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Yang, Yiping

Overview:

The goal of Dr. Yang’s laboratory is to understand the molecular and cellular mechanisms leading to the generation of potent and long-lasting anti-tumor immunity, and to develop effective gene immunotherapeutic strategies for treating cancer. Furthermore, rational pre-clinical approaches will be tested in clinical trials in patients with Epstein-Barr virus (EBV)-related malignancies. Specifically, we focus on the following areas:

1. Innate Immunity to Viruses. Recombinant vaccinia virus and adenovirus have been developed as potent vaccine vehicles for treating cancer and infectious diseases. Recent studies have shown that the unique potency of these viruses lies in their effective activation of the innate immune system. How these viruses activate the innate immune system remains largely unknown. We have been interested in the role of pattern-recognition receptors including Toll-like receptors (TLRs)in innate immune recognition of these viruses as well as their signaling pathways. In addition, we are investigating the role of innate immune cells such as natural killer (NK) cells in innate and adaptive immune responses to these viruses. A full understanding of these processes will help us design effective vaccine strategies.

2. T Cell Memory. Eliciting long-lived memory T cell response is an ultimate goal of vaccination to provide long-term immunity against cancer. However, it is not clear what controls the formation of long-lived memory T cells. The understanding of mechanisms underlying memory T cell formation will provide important insights into the design of effective vaccines for treating cancer.

3. Regulatory T Cell Biology. Accumulating evidence has shown that the immunosuppressive CD4+CD25+Foxp3+ regulatory T cells (TReg) play a critical role in the suppression of anti-tumor immunity. However, little is known about how TReg suppress T cell activation in vivo. Delineation of mechanisms underlying TReg-mediated suppression in vivo will help develop strategies to overcome TReg-mediated suppression in favor of boosting anti-tumor immunity.

4. Immunotherapy for EBV-associated Malignancies. Clinically, EBV-associated malignancies such as Hodgkin’s lymphoma offer a unique model to explore antigen-defined immunotherapy approaches because EBV-derived tumor antigens are specific for tumor cells only. Using this clinical model, we will test the utility of rational strategies identified in our preclinical models.

Positions:

Professor of Medicine

Medicine, Hematologic Malignancies and Cellular Therapy
School of Medicine

Professor of Immunology

Immunology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

M.D. 1985

M.D. — Zhejiang University (China)

Ph.D. 1993

Ph.D. — University of Michigan at Ann Arbor

Residency, General Internal Medicine

University of Pennsylvania School of Medicine

Fellowship, Medical Oncology

Johns Hopkins University School of Medicine

Grants:

Medical Scientist Training Program

Administered By
School of Medicine
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1997
End Date
June 30, 2022

Translational Research in Surgical Oncology

Administered By
Surgery, Surgical Sciences
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
January 01, 2002
End Date
August 31, 2021

Role of hedgehog signaling in tumor-associated macrophage polarization

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 2017
End Date
June 30, 2021

Transfusion Medicine and Hematology

Administered By
Medicine, Hematology
AwardedBy
National Institutes of Health
Role
Preceptor
Start Date
July 01, 1975
End Date
June 30, 2021

T memory stem cells in cancer

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
December 01, 2015
End Date
November 30, 2020

Organization and Function of Cellular Structure

Administered By
Basic Science Departments
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1975
End Date
June 30, 2020

Basic Immunology Training Program

Administered By
Immunology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 2002
End Date
June 30, 2019

Viral Oncology Training Grant

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Participating Faculty Member
Start Date
July 01, 1980
End Date
June 30, 2019

Novel Strategies for Cancer Immunotherapy in Stem Cell Transplant

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
June 01, 2014
End Date
May 31, 2019

Smartphone Enabled Point-of-Care Detection of Serum Markers of Liver Cancer

Administered By
Biomedical Engineering
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
June 01, 2017
End Date
April 30, 2019

Role of Endogenous Toll-Like Receptor Ligands in Allospecific T Cell Activation

Administered By
Surgery, Abdominal Transplant Surgery
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 2012
End Date
December 31, 2017

Role of inflammation in cancer progression

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 01, 2014
End Date
July 31, 2017

Cancer Biology Training Grant

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Cancer Institute
Role
Mentor
Start Date
July 01, 1993
End Date
March 31, 2016

Mechanisms for NK Cell Activation in Vaccinia Virus Control

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
May 01, 2009
End Date
September 30, 2015

Novel Strategies for Cancer Immunotherapy in Stem Cell Transplant

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
March 01, 2009
End Date
May 31, 2014

IMVC Program Equipment Application: "BioRad CFX Connect Real-Time PCR Detection System"

Administered By
Medicine, Hematological Malignancies
AwardedBy
Georgia Regents University
Role
Principal Investigator
Start Date
May 01, 2013
End Date
April 30, 2014

TLR Signaling in CD8 T Cell Response to Vaccinia Virus

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
April 01, 2009
End Date
March 31, 2014

The Role of Toll-like Receptors in Cancer Immunotherapy

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 01, 2005
End Date
April 30, 2013

Multispectral Imaging Flow Cytometer Core

Administered By
Medicine, Medical Oncology
AwardedBy
National Institutes of Health
Role
Major User
Start Date
March 04, 2010
End Date
March 03, 2011

Immunotherapy for Epstein-Barr virus-associated malignancies

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 12, 2002
End Date
August 31, 2005
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Publications:

IL-18-dependent NKG2D ligand upregulation on accessory cells is mediated by the PI3K/GSK-3 pathway.

NK cells are critical for the control of viral infections. Studies have shown that efficient NK cell activation in response to infection with VV in vivo requires multiple pathways, including the NKG2D pathway. We have recently shown that IL-18 is necessary for the activation of NK cells through upregulation of the NKG2D ligand Rae-1 on DCs upon VV infection. However, how IL-18R signaling on the accessory cells contributes to Rae-1 up-regulation remains to be defined. In this study, we found IL-18-mediated Rae-1 up-regulation in accessory cells, including macrophages and DCs, to be dependent on the MyD88-PI3K pathway. We further found that IL-18 signaling through PI3K led to inhibition of GSK-3, which we found to be a negative regulator of Rae-1. Finally, we demonstrated that in vivo inhibition of GSK-3 could restore Rae-1 up-regulation on IL18R-/- DCs and partially rescue NK-cell activation against VV, leading to improved viral control in IL-18R-/- mice. Our results showed that IL18-dependent Rae-1 up-regulation on accessory cells is mediated by the MyD88-PI3K-GSK3 pathway. These observations may provide important insights into the design of effective NK cell-based immunotherapies.

Authors
Brandstadter, JD; Chen, H; Jiang, S; Huang, X; Yang, Y
MLA Citation
Brandstadter, JD, Chen, H, Jiang, S, Huang, X, and Yang, Y. "IL-18-dependent NKG2D ligand upregulation on accessory cells is mediated by the PI3K/GSK-3 pathway." Journal of leukocyte biology 101.6 (June 2017): 1317-1323.
PMID
28283665
Source
epmc
Published In
Journal of leukocyte biology
Volume
101
Issue
6
Publish Date
2017
Start Page
1317
End Page
1323
DOI
10.1189/jlb.2a0816-342r

Monocytic myeloid-derived suppressor cells regulate T-cell responses against vaccinia virus.

Vaccinia virus (VV) can potently activate NK- and T-cell responses, leading to efficient viral control and generation of long-lasting protective immunity. However, immune responses against viral infections are often tightly controlled to avoid collateral damage and systemic inflammation. We have previously shown that granulocytic myeloid-derived suppressor cells (g-MDSCs) can suppress the NK-cell response to VV infection. It remains unknown what regulates T-cell responses to VV infection in vivo. In this study, we first showed that monocytic MDSCs (m-MDSCs), but not g-MDSCs, from VV-infected mice could directly suppress CD4+ and CD8+ T-cell activation in vitro. We then demonstrated that defective recruitment of m-MDSCs to the site of VV infection in CCR2-/- mice enhanced VV-specific CD8+ T-cell response and that adoptive transfer of m-MDSCs into VV-infected mice suppressed VV-specific CD8+ T-cell activation, leading to a delay in viral clearance. Mechanistically, we further showed that T-cell suppression by m-MDSCs is mediated by indication of iNOS and production of NO upon VV infection, and that IFN-γ is required for activation of m-MDSCs. Collectively, our results highlight a critical role for m-MDSCs in regulating T-cell responses against VV infection and may suggest potential strategies using m-MDSCs to modulate T-cell responses during viral infections.

Authors
Fortin, C; Yang, Y; Huang, X
MLA Citation
Fortin, C, Yang, Y, and Huang, X. "Monocytic myeloid-derived suppressor cells regulate T-cell responses against vaccinia virus." European journal of immunology 47.6 (June 2017): 1022-1031.
PMID
28383204
Source
epmc
Published In
European Journal of Immunology
Volume
47
Issue
6
Publish Date
2017
Start Page
1022
End Page
1031
DOI
10.1002/eji.201646797

PD-L1 serves as a double agent in separating GVL from GVHD.

Allogeneic hematopoietic cell transplantation (HCT) represents a potentially curative treatment for a variety of hematologic malignancies due to the well-recognized graft-versus-leukemia/lymphoma (GVL) effect that is mediated by donor-derived alloreactive T cells. However, graft-versus-host disease (GVHD) is mediated by the same T cells and remains a significant clinical problem associated with substantial morbidity and mortality. In this issue of the JCI, Ni and colleagues used several murine models of GVHD to evaluate the effect of CD4+ T cell depletion on GVL versus GVHD and revealed that depletion of CD4+ T cells leads to the upregulation of PD-L1 by recipient tissues and donor CD8+ T cells. Interaction of PD-L1 with PD-1 in GVHD-targeted tissues resulted in CD8+ T cell exhaustion and apoptosis, thereby preventing GVHD, whereas PD-L1 interactions with CD80 in lymphoid tissue promoted CD8+ T cell survival and expansion, thereby enhancing the GVL response. By separating these seemingly similar alloreactive T cell responses based on the context of interaction, the results of this study may lay the groundwork for the development of effective clinical strategies to enhance GVL while minimizing GVHD following allogeneic HCT.

Authors
Brennan, TV; Yang, Y
MLA Citation
Brennan, TV, and Yang, Y. "PD-L1 serves as a double agent in separating GVL from GVHD." Journal of Clinical Investigation 127.5 (May 2017): 1627-1630.
PMID
28414300
Source
epmc
Published In
Journal of Clinical Investigation
Volume
127
Issue
5
Publish Date
2017
Start Page
1627
End Page
1630
DOI
10.1172/jci94196

IL-21 is required for CD4 memory formation in response to viral infection.

IL-21 has been shown to play an important role in the CD8 T cell response during acute and chronic viral infections. However, the role of IL-21 signaling in the CD4 T cell response to viral infection remains incompletely defined. In a model of infection with vaccinia virus, we show that intrinsic IL-21 signaling on CD4 T cells was critical for the formation of memory CD4 T cells in vivo. We further reveal that IL-21 promoted CD4 T cell survival in a mechanism dependent on activation of the STAT1 and STAT3 signaling pathways. In addition, the activation of Akt is also required for IL-21-dependent survival of CD4 T cells in vivo. These results identify a critical role for intrinsic IL-21 signaling in CD4 T cell survival and memory formation in response to viral infection in vivo and may provide insights into the design of effective vaccine strategies.

Authors
Yuan, Y; Yang, Y; Huang, X
MLA Citation
Yuan, Y, Yang, Y, and Huang, X. "IL-21 is required for CD4 memory formation in response to viral infection." JCI insight 2.7 (April 6, 2017): e90652-.
PMID
28405614
Source
epmc
Published In
JCI insight
Volume
2
Issue
7
Publish Date
2017
Start Page
e90652
DOI
10.1172/jci.insight.90652

Tumor-associated macrophages: implications in cancer immunotherapy.

Tumor-associated macrophages (TAMs), representing most of the leukocyte population in solid tumors, demonstrate great phenotypic heterogeneity and diverse functional capabilities under the influence of the local tumor microenvironment. These anti-inflammatory and protumorigenic macrophages modulate the local microenvironment to facilitate tumor growth and metastasis. In this review, we examine the origin of TAMs and the complex regulatory networks within the tumor microenvironment that facilitate the polarization of TAMs toward a protumoral phenotype. More extensively, we evaluate the mechanisms by which TAMs mediate angiogenesis, metastasis, chemotherapeutic resistance and immune evasion. Lastly, we will highlight novel interventional strategies targeting TAMs in preclinical studies and in early clinical trials that have significant potential in improving efficacy of current chemotherapeutic and/or immunotherapeutic approaches.

Authors
Petty, AJ; Yang, Y
MLA Citation
Petty, AJ, and Yang, Y. "Tumor-associated macrophages: implications in cancer immunotherapy." Immunotherapy 9.3 (March 2017): 289-302. (Review)
PMID
28231720
Source
epmc
Published In
Immunotherapy
Volume
9
Issue
3
Publish Date
2017
Start Page
289
End Page
302
DOI
10.2217/imt-2016-0135

Mechanisms of heparanase inhibitors in cancer therapy.

Heparanase is an endo-β-D-glucuronidase capable of cleaving heparan sulfate side chains contributing to breakdown of the extracellular matrix. Increased expression of heparanase has been observed in numerous malignancies and is associated with a poor prognosis. It has generated significant interest as a potential antineoplastic target because of the multiple roles it plays in tumor growth and metastasis. The protumorigenic effects of heparanase are enhanced by the release of heparan sulfate side chains, with subsequent increase in bioactive fragments and cytokine levels that promote tumor invasion, angiogenesis, and metastasis. Preclinical experiments have found heparanase inhibitors to substantially reduce tumor growth and metastasis, leading to clinical trials with heparan sulfate mimetics. In this review, we examine the role of heparanase in tumor biology and its interaction with heparan surface proteoglycans, specifically syndecan-1, as well as the mechanism of action for heparanase inhibitors developed as antineoplastic therapeutics.

Authors
Heyman, B; Yang, Y
MLA Citation
Heyman, B, and Yang, Y. "Mechanisms of heparanase inhibitors in cancer therapy." Experimental hematology 44.11 (November 2016): 1002-1012. (Review)
PMID
27576132
Source
epmc
Published In
Experimental Hematology
Volume
44
Issue
11
Publish Date
2016
Start Page
1002
End Page
1012
DOI
10.1016/j.exphem.2016.08.006

Ly6C(hi) monocytes regulate T cell responses in viral hepatitis.

Viral hepatitis remains a global health challenge despite recent progress in the development of more effective therapies. Although virus-specific CD8(+) and CD4(+) T cell responses are essential for viral clearance, it remains largely unknown what regulates T cell-mediated viral clearance. Thus, a better understanding of the regulation of anti-viral T cell immunity would be critical for the design of more effective therapies for viral hepatitis. Using a model of adenovirus-induced hepatitis, here we showed that adenoviral infection induced recruitment of Ly6C(hi) monocytes to the liver in a CCR2-dependent manner. These recruited Ly6C(hi) monocytes suppressed CD8(+) and CD4(+) T cell responses to adenoviral infection, leading to a delay in viral clearance. In vivo depletion of Ly6C(hi) monocytes markedly enhanced anti-viral T cell responses and promoted viral clearance. Mechanistically, we showed that induction of iNOS and the production of NO by Ly6C(hi) monocytes are critical for the suppression of T cell responses. In addition, a contact-dependent mechanism mediated by PD-1 and PD-L1 interaction is also required for T cell suppression by Ly6C(hi) monocytes. These findings suggest a critical role for Ly6C(hi) monocytes in the regulation of T cell immunity in viral hepatitis and may provide new insights into development of more effective therapies for treating viral hepatitis based on targeting the immunosuppressing monocytes.

Authors
Zhu, J; Chen, H; Huang, X; Jiang, S; Yang, Y
MLA Citation
Zhu, J, Chen, H, Huang, X, Jiang, S, and Yang, Y. "Ly6C(hi) monocytes regulate T cell responses in viral hepatitis." JCI insight 1.17 (October 20, 2016): e89880-.
PMID
27777980
Source
epmc
Published In
JCI insight
Volume
1
Issue
17
Publish Date
2016
Start Page
e89880

Driving an improved CAR for cancer immunotherapy.

The recent clinical success of chimeric antigen receptor (CAR) T cell therapy for B cell malignancies represents a paradigm shift in cancer immunotherapy. Unfortunately, application of CAR T cell-mediated therapy for solid tumors has so far been disappointing, and the reasons for this poor response in solid tumors remain unknown. In this issue of the JCI, Cherkassky and colleagues report on their use of a murine model of human pleural mesothelioma to explore potential factors that limit CAR T cell efficacy. Their studies have uncovered the importance of the tumor microenvironment in the inhibition of CAR T cell functions, revealed a critical role for the programmed death-1 (PD-1) pathway in CAR T cell exhaustion within the tumor microenvironment, and demonstrated improved antitumor effects with a CAR T cell-intrinsic PD-1 blockade strategy using a dominant negative form of PD-1. Together, the results of this study lay the groundwork for further evaluation of mechanisms underlying CAR T cell immune evasion within the tumor microenvironment for the improvement of CAR T cell-mediated therapy for solid tumors.

Authors
Huang, X; Yang, Y
MLA Citation
Huang, X, and Yang, Y. "Driving an improved CAR for cancer immunotherapy." The Journal of clinical investigation 126.8 (August 2016): 2795-2798.
PMID
27454296
Source
epmc
Published In
Journal of Clinical Investigation
Volume
126
Issue
8
Publish Date
2016
Start Page
2795
End Page
2798
DOI
10.1172/jci88959

Heparan sulfate mimetic PG545-mediated antilymphoma effects require TLR9-dependent NK cell activation.

Heparan sulfate (HS) is an essential component of the extracellular matrix (ECM), which serves as a barrier to tumor invasion and metastasis. Heparanase promotes tumor growth by cleaving HS chains of proteoglycan and releasing HS-bound angiogenic growth factors and facilitates tumor invasion and metastasis by degrading the ECM. HS mimetics, such as PG545, have been developed as antitumor agents and are designed to suppress angiogenesis and metastasis by inhibiting heparanase and competing for the HS-binding domain of angiogenic growth factors. However, how PG545 exerts its antitumor effect remains incompletely defined. Here, using murine models of lymphoma, we determined that the antitumor effects of PG545 are critically dependent on NK cell activation and that NK cell activation by PG545 requires TLR9. We demonstrate that PG545 does not activate TLR9 directly but instead enhances TLR9 activation through the elevation of the TLR9 ligand CpG in DCs. Specifically, PG545 treatment resulted in CpG accumulation in the lysosomal compartment of DCs, leading to enhanced production of IL-12, which is essential for PG545-mediated NK cell activation. Overall, these results reveal that PG545 activates NK cells and that this activation is critical for the antitumor effect of PG545. Moreover, our findings may have important implications for improving NK cell-based antitumor therapies.

Authors
Brennan, TV; Lin, L; Brandstadter, JD; Rendell, VR; Dredge, K; Huang, X; Yang, Y
MLA Citation
Brennan, TV, Lin, L, Brandstadter, JD, Rendell, VR, Dredge, K, Huang, X, and Yang, Y. "Heparan sulfate mimetic PG545-mediated antilymphoma effects require TLR9-dependent NK cell activation." The Journal of clinical investigation 126.1 (January 2016): 207-219.
PMID
26649979
Source
epmc
Published In
Journal of Clinical Investigation
Volume
126
Issue
1
Publish Date
2016
Start Page
207
End Page
219
DOI
10.1172/jci76566

Targeting the Human Notch 2-BCR Axis: A Driver of B-Cell Hyper-Responsiveness in Active Chronic Graft-Versus Host Disease (cGVHD)

Authors
Poe, JC; Jia, W; Li, Z; Hakim, FT; Pavletic, SZ; Rose, JJ; Rizzieri, DA; Yang, Y; Chen, BJ; Green, M; Anand, S; Siebel, CW; Maillard, I; Chao, NJ; Sarantopoulos, S
MLA Citation
Poe, JC, Jia, W, Li, Z, Hakim, FT, Pavletic, SZ, Rose, JJ, Rizzieri, DA, Yang, Y, Chen, BJ, Green, M, Anand, S, Siebel, CW, Maillard, I, Chao, NJ, and Sarantopoulos, S. "Targeting the Human Notch 2-BCR Axis: A Driver of B-Cell Hyper-Responsiveness in Active Chronic Graft-Versus Host Disease (cGVHD)." December 3, 2015.
Source
wos-lite
Published In
Blood
Volume
126
Issue
23
Publish Date
2015

Cancer immunotherapy: harnessing the immune system to battle cancer.

The recent clinical successes of immune checkpoint blockade and chimeric antigen receptor T cell therapies represent a turning point in cancer immunotherapy. These successes also underscore the importance of understanding basic tumor immunology for successful clinical translation in treating patients with cancer. The Reviews in this Review Series focus on current developments in cancer immunotherapy, highlight recent advances in our understanding of basic aspects of tumor immunology, and suggest how these insights can lead to the development of new immunotherapeutic strategies.

Authors
Yang, Y
MLA Citation
Yang, Y. "Cancer immunotherapy: harnessing the immune system to battle cancer." The Journal of clinical investigation 125.9 (September 2015): 3335-3337. (Review)
PMID
26325031
Source
epmc
Published In
Journal of Clinical Investigation
Volume
125
Issue
9
Publish Date
2015
Start Page
3335
End Page
3337
DOI
10.1172/jci83871

Innate immune activation by tissue injury and cell death in the setting of hematopoietic stem cell transplantation.

Allogeneic hematopoietic stem cell transplantation (Allo-HSCT) with donor lymphocyte infusion is the mainstay of treatment for many types of hematological malignancies, but the therapeutic effect and prevention of relapse is complicated by donor T-cell recognition and attack of host tissue in a process known as graft-versus-host disease (GvHD). Cytotoxic myeloablative conditioning regimens used prior to Allo-HSCT result in the release of endogenous innate immune activators that are increasingly recognized for their role in creating a pro-inflammatory milieu. This increased inflammatory state promotes allogeneic T-cell activation and the induction and perpetuation of GvHD. Here, we review the processes of cellular response to injury and cell death that are relevant following Allo-HSCT and present the current evidence for a causative role of a variety of endogenous innate immune activators in the mediation of sterile inflammation following Allo-HSCT. Finally, we discuss the potential therapeutic strategies that target the endogenous pathways of innate immune activation to decrease the incidence and severity of GvHD following Allo-HSCT.

Authors
Brennan, TV; Rendell, VR; Yang, Y
MLA Citation
Brennan, TV, Rendell, VR, and Yang, Y. "Innate immune activation by tissue injury and cell death in the setting of hematopoietic stem cell transplantation." Frontiers in immunology 6 (January 2015): 101-. (Review)
PMID
25852683
Source
epmc
Published In
Frontiers in Immunology
Volume
6
Publish Date
2015
Start Page
101
DOI
10.3389/fimmu.2015.00101

NK cell-extrinsic IL-18 signaling is required for efficient NK-cell activation by vaccinia virus.

NK cells are important for the control of vaccinia virus (VV) in vivo. Recent studies have shown that multiple pathways are required for effective activation of NK cells. These include both TLR-dependent and -independent pathways, as well as the NKG2D activating receptor that recognizes host stress-induced NKG2D ligands. However, it remains largely unknown what controls the upregulation of NKG2D ligands in response to VV infection. In this study using C57BL/6 mice, we first showed that IL-18 is critical for NK-cell activation and viral clearance. We then demonstrated that IL-18 signaling on both NK cells and DCs is required for efficient NK-cell activation upon VV infection in vitro. We further showed in vivo that efficient NK-cell activation in response to VV is dependent on DCs and IL-18 signaling in non-NK cells, suggesting an essential role for NK cell-extrinsic IL-18 signaling in NK-cell activation. Mechanistically, IL-18 signaling in DCs promotes expression of Rae-1, an NKG2D ligand. Collectively, our data reveal a previously unrecognized role for NK cell-extrinsic IL-18 signaling in NK-cell activation through upregulation of NKG2D ligands. These observations may provide insights into the design of effective NK-cell-based therapies for viral infections and cancer.

Authors
Brandstadter, JD; Huang, X; Yang, Y
MLA Citation
Brandstadter, JD, Huang, X, and Yang, Y. "NK cell-extrinsic IL-18 signaling is required for efficient NK-cell activation by vaccinia virus." European journal of immunology 44.9 (September 2014): 2659-2666.
PMID
24846540
Source
epmc
Published In
European Journal of Immunology
Volume
44
Issue
9
Publish Date
2014
Start Page
2659
End Page
2666
DOI
10.1002/eji.201344134

Both NK cell-intrinsic and -extrinsic STAT1 signaling are required for NK cell response against vaccinia virus.

NK cells play an important role in innate immune control of the infection with vaccinia virus (VV). However, it remains incompletely defined how the activation of NK cells in response to VV is regulated. In this study, we showed that STAT1 was critical for NK cell activation upon VV infection and the subsequent clearance of VV infection in vivo. We further demonstrated that STAT1 signaling in both NK and accessory cells such as dendritic cells was required for efficient NK cell activation upon VV infection. Mechanistically, STAT1 signaling in dendritic cells promoted the expression of NKG2D ligands, which is required for NK cell activation via the NKG2D pathway. Taken together, our data suggest that STAT1 mediates anti-VV effect by promoting NK cell activation through both NK-intrinsic and extrinsic mechanisms and may provide insights into the design of effective NK cell-based therapies for viral infections.

Authors
Fortin, C; Huang, X; Yang, Y
MLA Citation
Fortin, C, Huang, X, and Yang, Y. "Both NK cell-intrinsic and -extrinsic STAT1 signaling are required for NK cell response against vaccinia virus." J Immunol 191.1 (July 1, 2013): 363-368.
PMID
23733873
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
191
Issue
1
Publish Date
2013
Start Page
363
End Page
368
DOI
10.4049/jimmunol.1202714

A mouse model for cancer immunoediting with renal cell carcinoma to explore mechanisms of immune escape

Authors
Bigger, E; Quigley, M; Yang, Y
MLA Citation
Bigger, E, Quigley, M, and Yang, Y. "A mouse model for cancer immunoediting with renal cell carcinoma to explore mechanisms of immune escape." May 20, 2013.
Source
wos-lite
Published In
Journal of Clinical Oncology
Volume
31
Issue
15
Publish Date
2013

Jak3 Dependent Inhibition of Alloreactive T Cell Proliferation in the Setting of Chronic TLR Stimulation.

Authors
Brennan, T; Lin, L; Yang, Y
MLA Citation
Brennan, T, Lin, L, and Yang, Y. "Jak3 Dependent Inhibition of Alloreactive T Cell Proliferation in the Setting of Chronic TLR Stimulation." April 2013.
Source
wos-lite
Published In
American Journal of Transplantation
Volume
13
Publish Date
2013
Start Page
256
End Page
256

Long-Term Persistence of Recipient MHC Class-II Antigen Following Allogeneic Bone Marrow Transplantation.

Authors
Brennan, T; Lin, L; Yang, Y
MLA Citation
Brennan, T, Lin, L, and Yang, Y. "Long-Term Persistence of Recipient MHC Class-II Antigen Following Allogeneic Bone Marrow Transplantation." April 2013.
Source
wos-lite
Published In
American Journal of Transplantation
Volume
13
Publish Date
2013
Start Page
375
End Page
375

The benefits of restraint: a pivotal role for IL-13 in hepatic glucose homeostasis.

In response to feeding, insulin promotes the uptake of sugar in peripheral tissues and suppresses the production of sugar, a process called gluconeogenesis, in the liver. Recent research has shown that chronic inflammation promotes insulin resistance, and in turn, chronically high glucose levels can drive inflammation. In this issue of the JCI, Stanya et al. investigate the connection between inflammation and glucose homeostasis by analyzing the effect of the antiinflammatory cytokine IL-13. Their results suggest that IL-13 plays an unexpected role in the regulation of glucose homeostasis by modulating gluconeogenesis and may be a useful therapeutic target for treatment of diabetes and metabolic syndrome.

Authors
Machado, MV; Yang, Y; Diehl, AM
MLA Citation
Machado, MV, Yang, Y, and Diehl, AM. "The benefits of restraint: a pivotal role for IL-13 in hepatic glucose homeostasis." J Clin Invest 123.1 (January 2013): 115-117.
PMID
23257364
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
123
Issue
1
Publish Date
2013
Start Page
115
End Page
117
DOI
10.1172/JCI67238

Survival analysis of tongue squamous cell carcinoma with CXCR4, CD44 and CD133 expression

Objective: This study aimed to analyze the correlation of the expression of CXCR4, CD44, and CD133 proteins with the clinicopathological characteristics of patients to identify the factors affecting the post-operation survival rate of tongue squamous cell carcinomas (TSCCs). Methods: Clinical data of 44 patients with TSCCs were collected and retrospectively analyzed. The diagnoses of all cases were pathologically confirmed. CXCR4, CD44, and CD133 expression in 44 TSCCs patients with different pathological grades was examined immunohistochemically. Survival curves were processed in accordance with the Kaplan-Meier method. The Cox regression model was used for the multivariate analysis of relevant clinical and survival data. Results: Among the 44 examined TSCCs patients, 29 cases were well differentiated and 15 were moderately or poor differentiated; 11 cases were stage I, 12 were stage II, 8 were stage III, and 13 were stage IV. Positive staining of CXCR4, CD44, and CD133 was found in all cases with different degrees. According to the pathological tumor grade, the positive rates of CXCR4, CD44, and CD133 expression were 79.54% (35/44 cases), 77.27% (34/44 cases), and 75.00% (33/44 cases), respectively. Expression of CXCR4, CD44, and CD133 significantly differed between different histological grades (P<0.05). Correlation analysis indicated that the expression of CXCR4, CD44, and CD133 was positively correlated with the metastasis, recurrence of TSCCs. COX multivariate analysis indicated that CXCR4 expression, clinical stage, and neck metastasis were independent prognostic predictors of TSCCs patients and risk factors of death. Conclusion: CXCR4, CD44, and CD133 may be correlated with the malignancy of TSCCs. CXCR4 expression, clinical stage, cervical lymph node metastasis were the correlated prognosis factors of TSCC patients after operation.

Authors
Nong, X; Xu, M; Li, H; Yang, Y; Nong, D; Cao, Y; Li, J; Xu, H; Li, Y
MLA Citation
Nong, X, Xu, M, Li, H, Yang, Y, Nong, D, Cao, Y, Li, J, Xu, H, and Li, Y. "Survival analysis of tongue squamous cell carcinoma with CXCR4, CD44 and CD133 expression." Chinese Journal of Clinical Oncology 40.14 (2013): 832-837.
Source
scival
Published In
Chinese Journal of Clinical Oncology
Volume
40
Issue
14
Publish Date
2013
Start Page
832
End Page
837
DOI
10.3969/j.issn.1000-8179.2013.14.005

Heparan sulfate, an endogenous TLR4 agonist, promotes acute GVHD after allogeneic stem cell transplantation.

Graft-versus-host disease (GVHD) remains the most common cause of nonrelapse-related morbidity and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Although T-cell depletion and intensive immunosuppression are effective in the control of GVHD, they are often associated with higher rates of infection and tumor recurrence. In this study, we showed that heparan sulfate (HS), an extracellular matrix component, can activate Toll-like receptor 4 on dendritic cells in vitro, leading to the enhancement of dendritic cell maturation and alloreactive T-cell responses. We further demonstrated in vivo that serum HS levels were acutely elevated at the onset of clinical GVHD in mice after allo-HSCT. Treatment with the serine protease inhibitor α1-antitrypsin decreased serum levels of HS, leading to a reduction in alloreactive T-cell responses and GVHD severity. Conversely, an HS mimetic that increased serum HS levels accelerated GVHD. In addition, in patients undergoing allo-HSCT for hematologic malignancies, serum HS levels were elevated and correlated with the severity of GVHD. These results identify a critical role for HS in promoting acute GVHD after allo-HSCT, and they suggest that modulation of HS release may have therapeutic potential for the control of clinical GVHD.

Authors
Brennan, TV; Lin, L; Huang, X; Cardona, DM; Li, Z; Dredge, K; Chao, NJ; Yang, Y
MLA Citation
Brennan, TV, Lin, L, Huang, X, Cardona, DM, Li, Z, Dredge, K, Chao, NJ, and Yang, Y. "Heparan sulfate, an endogenous TLR4 agonist, promotes acute GVHD after allogeneic stem cell transplantation." Blood 120.14 (October 4, 2012): 2899-2908.
PMID
22760779
Source
pubmed
Published In
Blood
Volume
120
Issue
14
Publish Date
2012
Start Page
2899
End Page
2908
DOI
10.1182/blood-2011-07-368720

The development and function of memory regulatory T cells after acute viral infections.

Natural CD4+CD25+Foxp3+ regulatory T cells (Tregs) are critical for the control of immune responses to pathogens. However, most studies have focused on chronic infections, in which pathogen-specific Tregs contribute to pathogen persistence and, in some cases, concomitant immunity. How Tregs behave and function following acute infections remains largely unknown. In this article, we show that pathogen-specific Tregs can be activated and expand upon acute viral infections in vivo. The activated Tregs then contract to form a memory pool after resolution of the infection. These memory Tregs expand rapidly upon a secondary challenge, secrete large amounts of IL-10, and suppress excessive immunopathological conditions elicited by recall expansion of non-Tregs via an IL-10-dependent mechanism. Our work reveals a memory Treg population that develops after acute viral infections and may help in the design of effective strategies to circumvent excessive immunopathological effects.

Authors
Sanchez, AM; Zhu, J; Huang, X; Yang, Y
MLA Citation
Sanchez, AM, Zhu, J, Huang, X, and Yang, Y. "The development and function of memory regulatory T cells after acute viral infections." J Immunol 189.6 (September 15, 2012): 2805-2814.
PMID
22855712
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
189
Issue
6
Publish Date
2012
Start Page
2805
End Page
2814
DOI
10.4049/jimmunol.1200645

NK cell response to vaccinia virus is regulated by myeloid-derived suppressor cells.

NK cells are critical for the innate immune control of poxviral infections. Previous studies have shown that NK cells are efficiently activated in response to infection with vaccinia virus (VV), the most studied member of the poxvirus family. However, it remains unknown whether the activation of NK cells in response to VV infection is tightly regulated. In this study, we showed that myeloid-derived suppressor cells (MDSCs) rapidly accumulated at the site of VV infection. In vivo depletion of MDSCs led to enhanced NK cell proliferation, activation, and function in response to VV infection. This was accompanied by an increase in mortality and systemic IFN-γ production. We further demonstrated that the granulocytic-MDSC (G-MDSC) subset was responsible for the suppression on NK cells and that this suppression was mediated by reactive oxygen species. These results indicate that G-MDSCs can negatively regulate NK cell activation and function in response to VV infection and suggest that manipulation of G-MDSCs could represent an attractive strategy for regulating NK cell activities for potential therapeutic benefits.

Authors
Fortin, C; Huang, X; Yang, Y
MLA Citation
Fortin, C, Huang, X, and Yang, Y. "NK cell response to vaccinia virus is regulated by myeloid-derived suppressor cells." J Immunol 189.4 (August 15, 2012): 1843-1849.
PMID
22798671
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
189
Issue
4
Publish Date
2012
Start Page
1843
End Page
1849
DOI
10.4049/jimmunol.1200584

Differential impact of inhibitory and activating Killer Ig-Like Receptors (KIR) on high-risk patients with myeloid and lymphoid malignancies undergoing reduced intensity transplantation from haploidentical related donors.

The impact of activating KIR (aKIR) and inhibitory KIR (iKIR) on OS, relapse-related mortality (RRM) and acute GVHD (aGVHD) was prospectively studied in 84 adults with high-risk hematologic malignancies receiving reduced intensity conditioning (RIC) T-cell depleted hematopoietic SCT (HSCT) from haploidentical related donors. In this clinical model, freedom from RRM is dependent on GVL effect. Patients were divided into myeloid (n=49) and lymphoid (n=35) malignancy groups. KIR-ligand and ligand-ligand models were studied in both GVH and rejection directions and statistically correlated with outcome measures. In the myeloid group, OS was higher (P=0.009) and RRM was lower (P=0.036) in patients missing HLA-C group2 ligand to donor iKIR. OS was higher if patients had >1 missing ligand (P=0.018). In lymphoid malignancy, missing ligand to donor KIR had no impact on OS or RRM. However, OS was better with donor aKIR 2DS2 (P=0.028). There was a trend towards shorter OS in recipient with KIR 2DS1, 2DS5 and 3DS1, although sample sizes were too small to provide inferential statistics. Findings in lymphoid malignancy patients should be further studied. These results suggest that the absence of appropriate HLA ligands in the recipient to donor iKIR may induce GVL without aGVHD in myeloid malignancy patients undergoing TCD-RIC transplants.

Authors
Chen, D-F; Prasad, VK; Broadwater, G; Reinsmoen, NL; DeOliveira, A; Clark, A; Sullivan, KM; Chute, JP; Horwitz, ME; Gasparetto, C; Long, GD; Yang, Y; Chao, NJ; Rizzieri, DA
MLA Citation
Chen, D-F, Prasad, VK, Broadwater, G, Reinsmoen, NL, DeOliveira, A, Clark, A, Sullivan, KM, Chute, JP, Horwitz, ME, Gasparetto, C, Long, GD, Yang, Y, Chao, NJ, and Rizzieri, DA. "Differential impact of inhibitory and activating Killer Ig-Like Receptors (KIR) on high-risk patients with myeloid and lymphoid malignancies undergoing reduced intensity transplantation from haploidentical related donors." Bone Marrow Transplant 47.6 (June 2012): 817-823.
PMID
22139069
Source
pubmed
Published In
Bone Marrow Transplantation
Volume
47
Issue
6
Publish Date
2012
Start Page
817
End Page
823
DOI
10.1038/bmt.2011.181

Alpha 1-Antitrypsin Therapy Reduces Alloreactive T Cell Activation and Decreases Graft-vs-Host Disease Following Hematopoietic Stem Cell Transplantation

Authors
Brennan, TV; Lin, L; Huang, X; Sudan, DL; Yang, Y
MLA Citation
Brennan, TV, Lin, L, Huang, X, Sudan, DL, and Yang, Y. "Alpha 1-Antitrypsin Therapy Reduces Alloreactive T Cell Activation and Decreases Graft-vs-Host Disease Following Hematopoietic Stem Cell Transplantation." May 2012.
Source
wos-lite
Published In
American Journal of Transplantation
Volume
12
Publish Date
2012
Start Page
292
End Page
292

Myeloid-derived suppressor cells regulate natural killer cell response to adenovirus-mediated gene transfer

The attendant innate and adaptive immune responses to viral vectors have posed a significant hurdle for clinical application of viral vector-mediated gene therapy. Previous studies have shown that natural killer (NK) cells play a critical role in innate immune elimination of adenoviral vectors in the liver. However, it is not clear how the NK cell response to adenoviral vectors is regulated. In this study, we identified a role for granulocytic myeloid-derived suppressor cells (G-MDSCs) in this process. We show that in vivo administration of adenoviral vectors results in rapid accumulation of G-MDSCs early during adenoviral infection. In vivo depletion of both MDSC populations, but not monocytic MDSCs (M-MDSCs) alone, resulted in accelerated clearance of adenoviral vectors in the liver. This was accompanied by enhanced NK cell proliferation and activation, suggesting a role for MDSCs, probably G-MDSCs, in suppressing NK cell activation and function in vivo. We further demonstrate in vitro that G-MDSCs, but not M-MDSCs, are responsible for the suppression of NK cell activation. In addition, we show that adenoviral infection activated G-MDSCs to produce higher levels of reactive oxygen species (ROS) and that G-MDSC-mediated suppression of NK cells is mediated by ROS, specifically, H2O2. This study demonstrates for the first time that the NK cell response to adenoviral vectors is negatively regulated by G-MDSCs and suggests that G-MDSC-based strategies could potentially improve the outcome of viral vector-mediated gene therapy. © 2012, American Society for Microbiology.

Authors
Zhu, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Huang, X, and Yang, Y. "Myeloid-derived suppressor cells regulate natural killer cell response to adenovirus-mediated gene transfer." Journal of Virology 86.24 (2012): 13689-13696.
PMID
23055553
Source
scival
Published In
Journal of virology
Volume
86
Issue
24
Publish Date
2012
Start Page
13689
End Page
13696
DOI
10.1128/JVI.01595-12

NKT cells are essential for innate immune control of vaccinia viral infection in vivo

Authors
Novy, P; Yang, Y
MLA Citation
Novy, P, and Yang, Y. "NKT cells are essential for innate immune control of vaccinia viral infection in vivo." JOURNAL OF IMMUNOLOGY 186 (April 2011).
Source
wos-lite
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
186
Publish Date
2011

The role of natural regulatory T cells in infection.

Naturally occurring regulatory T cells (T(Reg)) suppress multiple cell types of the immune system to maintain dominant tolerance to protect from autoimmunity, down-modulate anti-tumor immunity and restrain allergic diseases. In addition to these functions, T(Reg) can alter effector responses to invading pathogens, leading to a variety of outcomes affecting both the host and infecting microorganisms. Here, we review how T(Reg) can influence the immune responses to chronic infections where pathogen-specific T(Reg) can contribute to pathogen persistence and, in some cases, concomitant immunity, as well as control immunopathology associated with robust immune responses. We also review the data on T(Reg) during acute infection, focusing on the questions these studies raise regarding the most appropriate model(s) to examine T(Reg) during infection. Finally, we discuss the ways in which the T(Reg) function can be altered by invading pathogens and how these can be exploited to develop methods therapeutically to influence disease and vaccine outcomes.

Authors
Sanchez, AM; Yang, Y
MLA Citation
Sanchez, AM, and Yang, Y. "The role of natural regulatory T cells in infection." Immunol Res 49.1-3 (April 2011): 124-134. (Review)
PMID
21116872
Source
pubmed
Published In
Immunologic Research
Volume
49
Issue
1-3
Publish Date
2011
Start Page
124
End Page
134
DOI
10.1007/s12026-010-8176-8

Intrinsic IL-21 signaling is critical for CD8 T cell survival and memory formation in response to vaccinia viral infection.

CD4 T cell help plays an important role in promoting CD8 T cell immunity to pathogens. In models of infection with vaccinia virus (VV) and Listeria monocytogenes, CD4 T cell help is critical for the survival of activated CD8 T cells during both the primary and memory recall responses. Still unclear, however, is how CD4 T cell help promotes CD8 T cell survival. In this study, we first showed that CD4 T cell help for the CD8 T cell response to VV infection was mediated by IL-21, a cytokine produced predominantly by activated CD4 T cells, and that direct action of IL-21 on CD8 T cells was critical for the VV-specific CD8 T cell response in vivo. We next demonstrated that this intrinsic IL-21 signaling was essential for the survival of activated CD8 T cells and the generation of long-lived memory cells. We further revealed that IL-21 promoted CD8 T cell survival in a mechanism dependent on activation of the STAT1 and STAT3 pathways and subsequent upregulation of the prosurvival molecules Bcl-2 and Bcl-x(L). These results identify a critical role for intrinsic IL-21 signaling in CD8 T cell responses to an acute viral infection in vivo and may help design effective vaccine strategies.

Authors
Novy, P; Huang, X; Leonard, WJ; Yang, Y
MLA Citation
Novy, P, Huang, X, Leonard, WJ, and Yang, Y. "Intrinsic IL-21 signaling is critical for CD8 T cell survival and memory formation in response to vaccinia viral infection." J Immunol 186.5 (March 1, 2011): 2729-2738.
PMID
21257966
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
186
Issue
5
Publish Date
2011
Start Page
2729
End Page
2738
DOI
10.4049/jimmunol.1003009

Feasibility of low-dose interleukin-2 therapy following T-cell-depleted nonmyeloablative allogeneic hematopoietic stem cell transplantation from HLA-matched or -mismatched family member donors.

INTRODUCTION: High relapse rates and infections remain primary causes of failure in nonmyeloablative transplantation. Interleukin-2 (IL-2) may stimulate the immune system and improve outcomes. The primary objective of this pilot study was to evaluate the feasibility of administering IL-2 following a T-cell-depleted nonmyeloablative hematopoietic stem cell transplant. METHODS: Patients received T-cell-depleted nonmyeloablative transplant from a matched or mismatched related donor. Those with allogeneic engraftment,

Authors
Rizzieri, DA; Crout, C; Storms, R; Golob, J; Long, GD; Gasparetto, C; Sullivan, KM; Horwitz, M; Chute, J; Lagoo, AS; Morris, A; Beaven, A; Yang, Y; Peterson, B; Li, Z; Chao, NJ
MLA Citation
Rizzieri, DA, Crout, C, Storms, R, Golob, J, Long, GD, Gasparetto, C, Sullivan, KM, Horwitz, M, Chute, J, Lagoo, AS, Morris, A, Beaven, A, Yang, Y, Peterson, B, Li, Z, and Chao, NJ. "Feasibility of low-dose interleukin-2 therapy following T-cell-depleted nonmyeloablative allogeneic hematopoietic stem cell transplantation from HLA-matched or -mismatched family member donors." Cancer Invest 29.1 (January 2011): 56-61.
PMID
21166499
Source
pubmed
Published In
Cancer Investigation (Informa)
Volume
29
Issue
1
Publish Date
2011
Start Page
56
End Page
61
DOI
10.3109/07357907.2010.535055

Natural killer cell responses to viral infection.

Natural killer (NK) cells, as part of the innate immune system, play a key role in host defense against viral infections. Recent advances have indicated that NK cell activation and function are regulated by the interplay between inhibitory and activating signals. Thus, a better understanding of mechanisms responsible for NK cell activation and function in the control of viral infections will help develop NK cell-based therapies. In this review, we will first discuss how NK cells are activated in response to viral infections. We will then focus on the recruitment of activated NK cells to the site of infection as well as on NK cell effector mechanisms against virally infected cells.

Authors
Brandstadter, JD; Yang, Y
MLA Citation
Brandstadter, JD, and Yang, Y. "Natural killer cell responses to viral infection." J Innate Immun 3.3 (2011): 274-279. (Review)
PMID
21411975
Source
pubmed
Published In
Journal of Innate Immunity
Volume
3
Issue
3
Publish Date
2011
Start Page
274
End Page
279
DOI
10.1159/000324176

Targeting co-stimulatory pathways in gene therapy.

Gene therapy with recombinant viral vectors such as adenovirus and adenovirus-associated virus holds great promise in treating a wide range of diseases because of the high efficiency with which the viruses transfer their genomes into host cells in vivo. However, the activation of the host immune responses remains a major hurdle to successful gene therapy. Studies in the past two decades have elucidated the important role co-stimulation plays in the activation of both T and B cells. This review summarizes our current understanding of T cell co-stimulatory pathways, and strategies targeting these co-stimulatory pathways in gene therapy applications as well as potential future directions.

Authors
Huang, X; Yang, Y
MLA Citation
Huang, X, and Yang, Y. "Targeting co-stimulatory pathways in gene therapy. (Published online)" Front Microbiol 2 (2011): 202-.
PMID
22046171
Source
pubmed
Published In
Frontiers in Microbiology
Volume
2
Publish Date
2011
Start Page
202
DOI
10.3389/fmicb.2011.00202

NKG2D is required for NK cell activation and function in response to E1-deleted adenovirus.

Despite high transduction efficiency in vivo, the application of recombinant E1-deleted adenoviral vectors for in vivo gene therapy has been limited by the attendant innate and adaptive immune responses to adenoviral vectors. NK cells have been shown to play an important role in innate immune elimination of adenoviral vectors in vivo. However, the mechanisms underlying NK cell activation and function in response to adenoviral vectors remain largely undefined. In this study, we showed that NK cell activation upon adenoviral infection was dependent on accessory cells such as dendritic cells and macrophages and that cell contact-dependent signals from the accessory cells are necessary for NK cell activation. We further demonstrated that ligands of the NK activating receptor NKG2D were upregulated in accessory cells upon adenoviral infection and that blockade of NKG2D inhibited NK cell activation upon adenoviral infection, leading to a delay in adenoviral clearance in vivo. In addition, NKG2D was required for NK cell-mediated cytolysis on adenovirus-infected targets. Taken together, these results suggest that efficient NK cell activation and function in response to adenoviral infection is critically dependent on the NKG2D pathway, which understanding may assist in the design of effective strategies to improve the outcome of adenovirus-mediated gene therapy.

Authors
Zhu, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Huang, X, and Yang, Y. "NKG2D is required for NK cell activation and function in response to E1-deleted adenovirus." J Immunol 185.12 (December 15, 2010): 7480-7486.
PMID
21076062
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
185
Issue
12
Publish Date
2010
Start Page
7480
End Page
7486
DOI
10.4049/jimmunol.1002771

Natural killer cell-enriched donor lymphocyte infusions from A 3-6/6 HLA matched family member following nonmyeloablative allogeneic stem cell transplantation.

Infusing natural killer (NK) cells following transplantation may allow less infections and relapse with little risk of acute graft-versus-host disease (aGVHD). We delivered 51 total NK cell-enriched donor lymphocyte infusions (DLIs) to 30 patients following a 3-6/6 HLA matched T cell-depleted nonmyeloablative allogeneic transplant. The primary endpoint of this study was feasibility and safety. Eight weeks following transplantation, donor NK cell-enriched DLIs were processed using a CD56(+) selecting column with up to 3 fresh infusions allowed. Toxicity, relapse, and survival were monitored. T cell phenotype, NK cell functional recovery, and KIR typing were assessed for association with outcomes. Fourteen matched and 16 mismatched transplanted patients received a total of 51 NK cell-enriched DLIs. Selection resulted in 96% (standard deviation [SD] 8%) purity and 83% (SD 21%) yield in the matched setting and 97% (SD 3%) purity and 77% (SD 24%) yield in the mismatched setting. The median number of CD3(-) CD56(+) NK cells infused was 10.6 (SD 7.91) x 10(6) cells/kg and 9.21 (SD 5.6) x 10(6) cells/kg, respectively. The median number of contaminating CD3(+)CD56(-) T cells infused was .53 (1.1) x 10(6) and .27 (.78) x 10(6) in the matched and mismatched setting, respectively. Only 1 patient each in the matched (n = 14) or mismatched (n = 16) setting experienced severe aGVHD with little other toxicity attributable to the infusions. Long-term responders with multiple NK cell-enriched infusions and improved T cell phenotypic recovery had improved duration of responses (p = .0045) and overall survival (OS) (P = .0058). A 1-step, high-yield process is feasible, and results in high doses of NK cells infused with little toxicity. NK cell-enriched DLIs result in improved immune recovery and outcomes for some. Future studies must assess whether the improved outcomes are the direct result of the high doses and improved NK cell function or other aspects of immune recovery.

Authors
Rizzieri, DA; Storms, R; Chen, D-F; Long, G; Yang, Y; Nikcevich, DA; Gasparetto, C; Horwitz, M; Chute, J; Sullivan, K; Hennig, T; Misra, D; Apple, C; Baker, M; Morris, A; Green, PG; Hasselblad, V; Chao, NJ
MLA Citation
Rizzieri, DA, Storms, R, Chen, D-F, Long, G, Yang, Y, Nikcevich, DA, Gasparetto, C, Horwitz, M, Chute, J, Sullivan, K, Hennig, T, Misra, D, Apple, C, Baker, M, Morris, A, Green, PG, Hasselblad, V, and Chao, NJ. "Natural killer cell-enriched donor lymphocyte infusions from A 3-6/6 HLA matched family member following nonmyeloablative allogeneic stem cell transplantation." Biol Blood Marrow Transplant 16.8 (August 2010): 1107-1114.
PMID
20188202
Source
pubmed
Published In
Biology of Blood and Marrow Transplantation
Volume
16
Issue
8
Publish Date
2010
Start Page
1107
End Page
1114
DOI
10.1016/j.bbmt.2010.02.018

Targeting the TLR9-MyD88 pathway in the regulation of adaptive immune responses.

IMPORTANCE OF THE FIELD: Toll-like receptors (TLRs) are innate immune receptors critical in the innate immune defense against invading pathogens. Recent advances also reveal a crucial role for TLRs in shaping adaptive immune responses, conferring a potential therapeutic value to their modulation in the treatment of diseases. AREAS COVERED IN THIS REVIEW: The aim of this review is to discuss TLR9, the TLR9-MyD88 signaling pathway and its role in regulation of adaptive immune responses, as well as potential therapeutic implications by targeting this pathway. WHAT THE READER WILL GAIN: This review shows that the TLR9-MyD88 signaling pathway plays a critical role in promoting adaptive immune responses and that modulation of this pathway may have enormous therapeutic potential in enhancing vaccine potency, controlling autoimmunity, as well as improving the outcome of viral-vector-mediated gene therapy. TAKE HOME MESSAGE: Although TLR9 agonists have been used as adjuvants for enhancing vaccine potency, further exploitation of the TLR9-MyD88 pathway and its dynamic interaction with the immune system in vivo is needed to provide more effective therapeutic inventions in the design of vaccines for infectious diseases, allergies and cancer, in the control of autoimmunity, as well as in the improvement of viral-vector-mediated gene therapy.

Authors
Huang, X; Yang, Y
MLA Citation
Huang, X, and Yang, Y. "Targeting the TLR9-MyD88 pathway in the regulation of adaptive immune responses." Expert Opin Ther Targets 14.8 (August 2010): 787-796. (Review)
PMID
20560798
Source
pubmed
Published In
Expert Opinion on Therapeutic Targets
Volume
14
Issue
8
Publish Date
2010
Start Page
787
End Page
796
DOI
10.1517/14728222.2010.501333

Toll-like receptor 8-mediated activation of murine plasmacytoid dendritic cells by vaccinia viral DNA.

Plasmacytoid dendritic cells (pDCs) play a critical role in antiviral immunity through their ability to produce large amounts of type I IFNs. Activation of pDCs upon viral infection has been shown to be dependent on MyD88 and mediated by Toll-like receptors (TLR) 7 and 9, which sense viral ssRNA and CpG DNA, respectively. In this study, we showed that murine pDC recognition of vaccinia virus (VV), a dsDNA virus, was MyD88-dependent but TLR9-independent. Using HEK293 cells transfected with murine TLR7 or TLR8 and a NF-kappaB luciferase reporter, we demonstrated that stimulation of TLR8-, but not TLR7-, transfected cells with either VV or VV DNA resulted in substantial NF-kappaB activation, and that siRNA-mediated knockdown of TLR8 expression in pDCs led to a complete ablation of VV-induced type I IFN production. We further identified that the VV genome was rich in poly(A)/T sequences, and synthetic poly(A) and poly T oligodeoxynucleotides were capable of activating pDCs in a TLR8-dependent manner. In vivo, TLR8-MyD88-dependent pDC activation played a critical role in innate immune control of VV infection. Collectively, our data are unique in demonstrating that TLR8 is required for sensing poly(A)/T-rich DNA in pDCs, and that murine TLR8 is functional in the context of a viral infection.

Authors
Martinez, J; Huang, X; Yang, Y
MLA Citation
Martinez, J, Huang, X, and Yang, Y. "Toll-like receptor 8-mediated activation of murine plasmacytoid dendritic cells by vaccinia viral DNA." Proc Natl Acad Sci U S A 107.14 (April 6, 2010): 6442-6447.
PMID
20308556
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
107
Issue
14
Publish Date
2010
Start Page
6442
End Page
6447
DOI
10.1073/pnas.0913291107

Intrinsic IL-21 signaling is critical for CD8 T cell memory formation in response to Vaccinia viral infection

Authors
Novy, P; Yang, Y
MLA Citation
Novy, P, and Yang, Y. "Intrinsic IL-21 signaling is critical for CD8 T cell memory formation in response to Vaccinia viral infection." JOURNAL OF IMMUNOLOGY 184 (April 1, 2010).
Source
wos-lite
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
184
Publish Date
2010

Direct TLR2 signaling is critical for NK cell activation and function in response to vaccinia viral infection.

Natural killer (NK) cells play an essential role in innate immune control of poxviral infections in vivo. However, the mechanism(s) underlying NK cell activation and function in response to poxviruses remains poorly understood. In a mouse model of infection with vaccinia virus (VV), the most studied member of the poxvirus family, we identified that the Toll-like receptor (TLR) 2-myeloid differentiating factor 88 (MyD88) pathway was critical for the activation of NK cells and the control of VV infection in vivo. We further showed that TLR2 signaling on NK cells, but not on accessory cells such as dendritic cells (DCs), was necessary for NK cell activation and that this intrinsic TLR2-MyD88 signaling pathway was required for NK cell activation and played a critical role in the control of VV infection in vivo. In addition, we showed that the activating receptor NKG2D was also important for efficient NK activation and function, as well as recognition of VV-infected targets. We further demonstrated that VV could directly activate NK cells via TLR2 in the presence of cytokines in vitro and TLR2-MyD88-dependent activation of NK cells by VV was mediated through the phosphatidylinositol 3-kinase (PI3K)-extracellular signal-regulated kinase (ERK) pathway. Taken together, these results represent the first evidence that intrinsic TLR signaling is critical for NK cell activation and function in the control of a viral infection in vivo, indicate that multiple pathways are required for efficient NK cell activation and function in response to VV infection, and may provide important insights into the design of effective strategies to combat poxviral infections.

Authors
Martinez, J; Huang, X; Yang, Y
MLA Citation
Martinez, J, Huang, X, and Yang, Y. "Direct TLR2 signaling is critical for NK cell activation and function in response to vaccinia viral infection. (Published online)" PLoS Pathog 6.3 (March 12, 2010): e1000811-.
Website
http://hdl.handle.net/10161/4596
PMID
20300608
Source
pubmed
Published In
PLoS pathogens
Volume
6
Issue
3
Publish Date
2010
Start Page
e1000811
DOI
10.1371/journal.ppat.1000811

Reply to Bauer et al.: Murine pDC recognition of vaccinia viral DNA is mediated by TLR8

Authors
Martinez, J; Yang, Y
MLA Citation
Martinez, J, and Yang, Y. "Reply to Bauer et al.: Murine pDC recognition of vaccinia viral DNA is mediated by TLR8." Proceedings of the National Academy of Sciences of the United States of America 107.36 (2010): E140-.
Source
scival
Published In
Proceedings of the National Academy of Sciences of USA
Volume
107
Issue
36
Publish Date
2010
Start Page
E140
DOI
10.1073/pnas.1009858107

The TLR9-MyD88 pathway is critical for adaptive immune responses to adeno-associated virus gene therapy vectors in mice.

Recombinant adeno-associated viruses (AAVs) have been used widely for in vivo gene therapy. However, adaptive immune responses to AAV have posed a significant hurdle in clinical application of AAV vectors. Recent advances have suggested a crucial role for innate immunity in shaping adaptive immune responses. How AAV activates innate immunity, and thereby promotes AAV-targeted adaptive immune responses, remains unknown. Here we show that AAV activates mouse plasmacytoid DCs (pDCs) via TLR9 to produce type I IFNs. In vivo, the TLR9-MyD88 pathway was crucial to the activation of CD8+ T cell responses to both the transgene product and the AAV capsid, leading to loss of transgene expression and the generation of transgene product-specific and AAV-neutralizing antibodies. We further demonstrate that TLR9-dependent activation of adaptive immunity targeting AAV was mediated by type I IFNs and that human pDCs could be activated in vitro to induce type I IFN production via TLR9. These results reveal an essential role for the TLR9-MyD88-type I IFN pathway in induction of adaptive immune responses to AAV and suggest that strategies that interfere with this pathway may improve the outcome of AAV-mediated gene therapy in humans.

Authors
Zhu, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Huang, X, and Yang, Y. "The TLR9-MyD88 pathway is critical for adaptive immune responses to adeno-associated virus gene therapy vectors in mice." J Clin Invest 119.8 (August 2009): 2388-2398.
PMID
19587448
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
119
Issue
8
Publish Date
2009
Start Page
2388
End Page
2398
DOI
10.1172/JCI37607

Induction of type I IFN is required for overcoming tumor-specific T-cell tolerance after stem cell transplantation.

Tumor-specific T-cell tolerance represents one major mechanism of tumor-induced immune evasion. Myeloablative chemotherapy with stem cell transplantation may offer the best chance of achieving a state of minimal residual disease and, thus, minimize tumor-induced immune evasion. However, studies have shown that tumor-specific T-cell tolerance persists after transplantation. Here, we showed that CD4(+)CD25(+) regulatory T (T(Reg)) cells play a critical role in tumor-specific CD8(+) T-cell tolerance after transplantation. Removal of T(Reg) cells from the donor lymphocyte graft did not overcome this tolerance because of rapid conversion of donor CD4(+)CD25(-) T cells into CD4(+)CD25(+)Foxp3(+) T(Reg) cells in recipients after transplantation, and depletion of T(Reg) cells in recipients was necessary for the reversal of tumor-specific tolerance. These results suggest that strategies capable of overcoming T-cell tolerance in recipients are required to promote antitumor immunity after transplantation. Toward this goal, we showed that dendritic cell (DC) vaccines coadministered with the TLR9 ligand, CpG could effectively overcome tumor-specific tolerance, leading to significant prolongation of tumor-free survival after transplantation. We further showed that CpG-induced type I interferon was critical for the reversal of tumor-specific tolerance in vivo. Collectively, these results may suggest effective immunotherapeutic strategies for treating cancer after stem cell transplantation.

Authors
Horkheimer, I; Quigley, M; Zhu, J; Huang, X; Chao, NJ; Yang, Y
MLA Citation
Horkheimer, I, Quigley, M, Zhu, J, Huang, X, Chao, NJ, and Yang, Y. "Induction of type I IFN is required for overcoming tumor-specific T-cell tolerance after stem cell transplantation." Blood 113.21 (May 21, 2009): 5330-5339.
PMID
19279333
Source
pubmed
Published In
Blood
Volume
113
Issue
21
Publish Date
2009
Start Page
5330
End Page
5339
DOI
10.1182/blood-2008-05-155150

Modulation of CD4(+)CD25(+)Foxp3(+) Regulatory T Cells in the Recipients Is Necessary for Enhancing Anti-Tumor Immunity Following Stem Cell Transplantation

Authors
Horkheimer, I; Quigley, M; Zhu, J; Huang, X; Chao, N; Yang, Y
MLA Citation
Horkheimer, I, Quigley, M, Zhu, J, Huang, X, Chao, N, and Yang, Y. "Modulation of CD4(+)CD25(+)Foxp3(+) Regulatory T Cells in the Recipients Is Necessary for Enhancing Anti-Tumor Immunity Following Stem Cell Transplantation." May 2009.
Source
wos-lite
Published In
Molecular Therapy
Volume
17
Publish Date
2009
Start Page
S209
End Page
S210

The TLR9-MyD88 Pathway Is Critical for Adaptive Immune Responses to AAV Vectors in Gene Therapy

Authors
Zhu, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Huang, X, and Yang, Y. "The TLR9-MyD88 Pathway Is Critical for Adaptive Immune Responses to AAV Vectors in Gene Therapy." May 2009.
Source
wos-lite
Published In
Molecular Therapy
Volume
17
Publish Date
2009
Start Page
S296
End Page
S296

Innate immune recognition of viruses and viral vectors.

Recombinant viral vectors such as adenovirus and adenovirus-associated virus have been used widely as vehicles for gene therapy applications because of the high efficiency with which they transfer genes into a wide spectrum of cells in vivo. However, enthusiasm for the use of viral vectors in gene therapy has been tempered by significant problems of attendant host cellular and humoral immune responses that limit their safety and efficacy in vivo. Advances in immunology have suggested a crucial role for the innate immune system in the induction of immune responses to viruses. Thus, a better understanding of the mechanisms by which the host's innate immune system recognizes viruses and viral vectors will help in the design of effective strategies to improve the outcome of viral vector-mediated gene therapy. In this review we first discuss our current understanding of innate immune recognition of viruses in general, and then focus on the innate immune responses to viral vectors for gene therapy.

Authors
Huang, X; Yang, Y
MLA Citation
Huang, X, and Yang, Y. "Innate immune recognition of viruses and viral vectors." Hum Gene Ther 20.4 (April 2009): 293-301. (Review)
PMID
19272012
Source
pubmed
Published In
Human Gene Therapy
Volume
20
Issue
4
Publish Date
2009
Start Page
293
End Page
301
DOI
10.1089/hum.2008.141

A critical role for direct TLR2-MyD88 signaling in CD8 T-cell clonal expansion and memory formation following vaccinia viral infection.

Recent advances have suggested a crucial role of the innate immunity in shaping adaptive immune responses. How activation of innate immunity promotes adaptive T-cell responses to pathogens in vivo is not fully understood. It has been thought that Toll-like receptor (TLR)-mediated control of adaptive T-cell responses is mainly achieved by the engagement of TLRs on antigen-presenting cells to promote their maturation and function. In this study, we showed that direct TLR2-myeloid differentiating factor 88 (MyD88) signaling in CD8 T cells was also required for their efficient clonal expansion by promoting the survival of activated T cells on vaccinia viral infection in vivo. Effector CD8 T cells that lacked direct TLR2-MyD88 signaling did not survive the contraction phase to differentiate into long-lived memory cells. Furthermore, we observed that direct TLR2 ligation on CD8 T cells promoted CD8 T-cell proliferation and survival in vitro in a manner dependent on the phosphatidylinositol 3-kinase (PI3K)-Akt pathway activation and that activation of Akt controlled memory cell formation in vivo. These results identify a critical role for intrinsic TLR2-MyD88 signaling and PI3K-Akt pathway activation in CD8 T-cell clonal expansion and memory formation in vivo and could lead to the development of new vaccine approaches.

Authors
Quigley, M; Martinez, J; Huang, X; Yang, Y
MLA Citation
Quigley, M, Martinez, J, Huang, X, and Yang, Y. "A critical role for direct TLR2-MyD88 signaling in CD8 T-cell clonal expansion and memory formation following vaccinia viral infection." Blood 113.10 (March 5, 2009): 2256-2264.
PMID
18948575
Source
pubmed
Published In
Blood
Volume
113
Issue
10
Publish Date
2009
Start Page
2256
End Page
2264
DOI
10.1182/blood-2008-03-148809

A critical role for type I IFN-dependent NK cell activation in innate immune elimination of adenoviral vectors in vivo.

Recombinant adenoviruses have been used widely for gene therapy due to their high transduction efficiency in vivo. However, the attendant innate immune response to adenoviral vectors has limited their applications for in vivo gene therapy. Recent studies have shown that adenoviruses activate the innate immunity through both Toll-like receptor-dependent (TLR-dependent) and TLR-independent pathways, leading to the production of type I interferons (IFNs) and other inflammatory cytokines. Furthermore, type I IFNs play a pivotal role in innate immune elimination of adenoviral vectors in vivo. It remains to be defined how type I IFNs regulate innate immune clearance of adenoviral vectors. In this study, we showed in vivo that natural killer (NK) cells were activated and accumulated in the liver upon intravenous administration of adenoviral vectors, leading to the loss of adenoviral genome and the reduction of transgene expression. We further demonstrated that type I IFNs were critical for the activation of NK cells. This was achieved by direct action of type I IFNs on NK cells. Overall, our observations reveal a critical role for type I IFN-dependent NK cell activation in innate immune elimination of adenoviral vectors in vivo and may help design effective strategies to improve the outcome of adenovirus-mediated gene therapy.

Authors
Zhu, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Huang, X, and Yang, Y. "A critical role for type I IFN-dependent NK cell activation in innate immune elimination of adenoviral vectors in vivo." Mol Ther 16.7 (July 2008): 1300-1307.
PMID
18443600
Source
pubmed
Published In
Molecular Therapy
Volume
16
Issue
7
Publish Date
2008
Start Page
1300
End Page
1307
DOI
10.1038/mt.2008.88

STAT1 signaling in CD8 T cells is required for their clonal expansion and memory formation following viral infection in vivo.

Recent advances have shown that direct type I IFN signaling on T cells is required for their efficient expansion in response to viral infections in vivo. It is not clear which intracellular signaling molecule is responsible for this effect. Although STAT1 has been shown to mediate many of the type I IFN-dependent biological effects, its role in T cells remains uncertain in vivo. In this study, we demonstrated that STAT1 signaling in CD8 T cells was required for their efficient expansion by promoting the survival of activated CD8 T cells upon vaccinia viral infection in vivo, suggesting that the direct effect of type I IFNs on CD8 T cells is mediated by STAT1. Furthermore, effector CD8 T cells that lack STAT1 signaling did not survive the contraction phase to differentiate into long-lived memory cells. These results identify a critical role for type I IFN-STAT1 signaling in multiple stages of CD8 T cell response in vivo and suggest that strategies to activate type I IFN-STAT1 signaling pathway may enhance vaccine potency.

Authors
Quigley, M; Huang, X; Yang, Y
MLA Citation
Quigley, M, Huang, X, and Yang, Y. "STAT1 signaling in CD8 T cells is required for their clonal expansion and memory formation following viral infection in vivo." J Immunol 180.4 (February 15, 2008): 2158-2164.
PMID
18250422
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
180
Issue
4
Publish Date
2008
Start Page
2158
End Page
2164

Direct action of type I IFN on NK cells is required for their activation in response to vaccinia viral infection in vivo.

Type I IFN plays an important role in the activation of NK cells. However, the mechanism underlying type I IFN-dependent NK cell activation remains largely unknown. A recent report suggested that type I IFN acted on accessory dendritic cells, leading to IL-15 production, and that subsequent trans-presentation of IL-15 was required for NK cell activation upon stimulation with synthetic TLR ligands. It is not clear how type I IFN regulates NK cell activation in response to live pathogens. Using a murine model of infection with vaccinia virus (VV), we previously demonstrated a critical role for type I IFN in the innate immune control of VV infection. In this study, we first showed that type I IFN did not directly protect L929 cells from VV infection in vitro and that type I IFN-dependent innate immune control of VV infection in vivo was mediated by activated NK cells. We further demonstrated that direct action of type I IFN on NK cells, but not on dendritic cells, is required for the activation of NK cells in response to VV infection both in vitro and in vivo, leading to efficient VV clearance. Our findings may help design effective strategies for the control of poxviral infections in vivo.

Authors
Martinez, J; Huang, X; Yang, Y
MLA Citation
Martinez, J, Huang, X, and Yang, Y. "Direct action of type I IFN on NK cells is required for their activation in response to vaccinia viral infection in vivo." J Immunol 180.3 (February 1, 2008): 1592-1597.
PMID
18209055
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
180
Issue
3
Publish Date
2008
Start Page
1592
End Page
1597

CD4 T cells are required for CD8 T cell survival during both primary and memory recall responses.

The role of CD4 T cell help in primary and secondary CD8 T cell responses to infectious pathogens remains incompletely defined. The primary CD8 T response to infections was initially thought to be largely independent of CD4 T cells, but it is not clear why some primary, pathogen-specific CD8 T cell responses are CD4 T cell dependent. Furthermore, although the generation of functional memory CD8 T cells is CD4 T cell help dependent, it remains controversial when the "help" is needed. In this study, we demonstrated that CD4 T cell help was not needed for the activation and effector differentiation of CD8 T cells during the primary response to vaccinia virus infection. However, the activated CD8 T cells showed poor survival without CD4 T cell help, leading to a reduction in clonal expansion and a diminished, but stable CD8 memory pool. In addition, we observed that CD4 T cell help provided during both the primary and secondary responses was required for the survival of memory CD8 T cells during recall expansion. Our study indicates that CD4 T cells play a crucial role in multiple stages of CD8 T cell response to vaccinia virus infection and may help to design effective vaccine strategies.

Authors
Novy, P; Quigley, M; Huang, X; Yang, Y
MLA Citation
Novy, P, Quigley, M, Huang, X, and Yang, Y. "CD4 T cells are required for CD8 T cell survival during both primary and memory recall responses." J Immunol 179.12 (December 15, 2007): 8243-8251.
PMID
18056368
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
179
Issue
12
Publish Date
2007
Start Page
8243
End Page
8251

Extent of stimulation controls the formation of memory CD8 T cells.

Only a small fraction of effector CD8 T cells survives to become long-lived memory cells, whereas the majority of them die after an acute infection. What controls the formation of memory CD8 T cells remains mostly unknown. In this study, we showed CD8 T cells primed earlier during vaccinia viral infection received stronger stimulation, divided more extensively, and survived better than those primed later, leading to generation of a larger memory pool. Despite differentiation into effectors, the late-primed CD8 T cells lacked full cell division, displayed increased apoptosis, and failed to develop into memory cells, suggesting that the extent of stimulation influences the survival of effector CD8 T cells. We further demonstrated that the extent of stimulation, which included both the duration and the levels of antigenic stimulation/costimulation, during priming determined the formation of memory CD8 T cells via controlling the extent of Akt activation, and functional suppression of Akt led to defective CD8 memory formation in vivo. Collectively, our data suggest that the extent of stimulation controls CD8 memory formation via activation of Akt and may provide important insights into the design of effective vaccines.

Authors
Quigley, M; Huang, X; Yang, Y
MLA Citation
Quigley, M, Huang, X, and Yang, Y. "Extent of stimulation controls the formation of memory CD8 T cells." J Immunol 179.9 (November 1, 2007): 5768-5777.
PMID
17947649
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
179
Issue
9
Publish Date
2007
Start Page
5768
End Page
5777

Innate immune response to adenoviral vectors is mediated by both Toll-like receptor-dependent and -independent pathways.

Recombinant adenoviral vectors have been widely used for gene therapy applications and as vaccine vehicles for treating infectious diseases such as human immunodeficiency virus disease. The innate immune response to adenoviruses represents the most significant hurdle in clinical application of adenoviral vectors for gene therapy, but it is an attractive feature for vaccine development. How adenovirus activates innate immunity remains largely unknown. Here we showed that adenovirus elicited innate immune response through the induction of high levels of type I interferons (IFNs) by both plasmacytoid dendritic cells (pDCs) and non-pDCs such as conventional DCs and macrophages. The innate immune recognition of adenovirus by pDCs was mediated by Toll-like receptor 9 (TLR9) and was dependent on MyD88, whereas that by non-pDCs was TLR independent through cytosolic sensing of adenoviral DNA. Furthermore, type I IFNs were pivotal in innate and adaptive immune responses to adenovirus in vivo, and type I IFN blockade diminished immune responses, resulting in more stable transgene expression and reduction of inflammation. These findings indicate that adenovirus activates innate immunity by its DNA through TLR-dependent and -independent pathways in a cell type-specific fashion, and they highlight a critical role for type I IFNs in innate and adaptive immune responses to adenoviral vectors. Our results that suggest strategies to interfere with type I IFN pathway may improve the outcome of adenovirus-mediated gene therapy, whereas approaches to activate the type I IFN pathway may enhance vaccine potency.

Authors
Zhu, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Huang, X, and Yang, Y. "Innate immune response to adenoviral vectors is mediated by both Toll-like receptor-dependent and -independent pathways." J Virol 81.7 (April 2007): 3170-3180.
PMID
17229689
Source
pubmed
Published In
Journal of virology
Volume
81
Issue
7
Publish Date
2007
Start Page
3170
End Page
3180
DOI
10.1128/JVI.02192-06

Type I IFN signaling on both B and CD4 T cells is required for protective antibody response to adenovirus.

Recombinant adenoviruses have been used as vehicles for gene therapy as well as vaccination against infectious diseases and cancer. Efficient activation of host B cell response to adenoviral vectors that leads to the generation of protective, neutralizing Ab, represents a major barrier for gene therapy, but an attractive feature for vaccine development. What regulate(s) potent B cell response to adenoviral vectors remains incompletely defined. In this study, we showed that type I IFNs induced upon adenoviral infection are critical for multiple stages of adaptive B cell response to adenovirus including early B cell activation, germinal center formation, Ig isotype switching as well as plasma cell differentiation. We further demonstrated that although type I IFN signaling on dendritic cells was important for the production of virus-specific IgM, the generation of protective neutralizing Ab critically depended on type I IFN signaling on both CD4 T and B cells. The results may suggest potential strategies for improving adenovirus-mediated gene therapy in vivo and/or the design of effective vaccines for cancer and infectious diseases.

Authors
Zhu, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Huang, X, and Yang, Y. "Type I IFN signaling on both B and CD4 T cells is required for protective antibody response to adenovirus." J Immunol 178.6 (March 15, 2007): 3505-3510.
PMID
17339445
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
178
Issue
6
Publish Date
2007
Start Page
3505
End Page
3510

Innate immunity against vaccinia virus is mediated by TLR2 and requires TLR-independent production of IFN-beta.

Vaccinia virus (VV) has been used extensively as a vaccine vehicle in the clinical application for infectious diseases and cancer. Previous studies have suggested that the unique potency of VV-based vaccine lies in its effective activation of the innate immune system. However, how VV activates innate immune pathways remains largely unknown. In this study, we showed that VV elicited innate immune response through both Toll-like receptor (TLR)-dependent and -independent pathways. The TLR pathway was mediated by TLR2 and MyD88, leading to the production of proinflammatory cytokines, whereas activation of the TLR-independent pathway resulted in the secretion of IFN-beta. More importantly, both TLR-dependent and -independent pathways were required for activating innate and adaptive immunity to VV in vivo. These findings represent the first evidence that innate immune recognition of VV is mediated by TLR2, demonstrate that one pathogen can target both TLR and non-TLR innate immune pathways to work together in achieving efficient activation of host defense, and suggest potential new strategies for the design of effective vaccines.

Authors
Zhu, J; Martinez, J; Huang, X; Yang, Y
MLA Citation
Zhu, J, Martinez, J, Huang, X, and Yang, Y. "Innate immunity against vaccinia virus is mediated by TLR2 and requires TLR-independent production of IFN-beta." Blood 109.2 (January 15, 2007): 619-625.
PMID
16973959
Source
pubmed
Published In
Blood
Volume
109
Issue
2
Publish Date
2007
Start Page
619
End Page
625
DOI
10.1182/blood-2006-06-027136

The fate of effector CD8 T cells in vivo is controlled by the duration of antigen stimulation.

What controls the fate of the T-cell response remains incompletely defined. Gain of effector function facilitated by costimulation has been thought to be a crucial factor in determining the outcome of the T-cell response, i.e. long-term memory in the presence of costimulation versus tolerance induction in the absence of costimulation. In this study, we show that while costimulation or cognate CD4 helps to promote the acquisition of effector function during the initial phase of the CD8 T-cell response, the fate of effector CD8 T cells is controlled by the duration of subsequent antigenic stimulation. Effector CD8 T cells differentiate into memory cells only after clearance of antigen, whereas in the presence of persistent antigen, effector CD8 T cells are tolerized. Furthermore, protective immunity against tumour cannot develop in the persisting antigen environment. These results suggest that removal of persisting antigen by other means might be a prerequisite for effective immunotherapy in cancer.

Authors
Huang, X; Yang, Y
MLA Citation
Huang, X, and Yang, Y. "The fate of effector CD8 T cells in vivo is controlled by the duration of antigen stimulation." Immunology 118.3 (July 2006): 361-371.
PMID
16827897
Source
pubmed
Published In
Immunology
Volume
118
Issue
3
Publish Date
2006
Start Page
361
End Page
371
DOI
10.1111/j.1365-2567.2006.02381.x

Protection against autoimmunity in nonlymphopenic hosts by CD4+ CD25+ regulatory T cells is antigen-specific and requires IL-10 and TGF-beta.

CD4+ CD25+ regulatory T cells (T(Reg)) play a critical role in the control of autoimmunity. However, little is known about how T(Reg) suppress self-reactive T cells in vivo, thus limiting the development of T(Reg)-based therapy for treating autoimmune diseases. This is in large part due to the dependency on a state of lymphopenia to demonstrate T(Reg)-mediated suppression in vivo and the unknown Ag specificity of T(Reg) in most experimental models. Using a nonlymphopenic model of autoimmune pneumonitis and T(Reg) with known Ag specificity, in this study we demonstrated that these T(Reg) can actively suppress activation of self-reactive T cells and protect mice from fatal autoimmune pneumonitis. The protection required T(Reg) with the same Ag specificity as the self-reactive T cells and depended on IL-10 and TGF-beta. These results suggest that suppression of autoimmunity by T(Reg) in vivo consists of multiple layers of regulation and advocate for a strategy involving Ag-specific T(Reg) for treating organ-specific autoimmunity, because they do not cause generalized immune suppression.

Authors
Huang, X; Zhu, J; Yang, Y
MLA Citation
Huang, X, Zhu, J, and Yang, Y. "Protection against autoimmunity in nonlymphopenic hosts by CD4+ CD25+ regulatory T cells is antigen-specific and requires IL-10 and TGF-beta." J Immunol 175.7 (October 1, 2005): 4283-4291.
PMID
16177068
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
175
Issue
7
Publish Date
2005
Start Page
4283
End Page
4291

Transient gain of effector function by CD8+ T cells undergoing peripheral tolerance to high-dose self-antigen.

Induction of peripheral T cell tolerance is mediated by bone marrow-derived dendritic cells that cross-present self-antigen to self-reactive T cells. The current model for peripheral CD8(+) T cell tolerance is that TCR engagement by self-antigen in the absence of costimulation results in abortive activation without development of effector function. Here we demonstrate in vivo that high-dose self-antigen ("signal 1") can compensate for lack of costimulation ("signal 2"), leading to full activation of and development of effector function by self-reactive T cells. In the setting of low-dose self-antigen, acquisition of effector function by self-reactive T cells is dependent on costimulation via CD40 ligation in vivo. However, gain of effector function in either setting does not prevent eventual tolerance of self-reactive CD8(+) T cells. These results suggest that the mechanisms for peripheral CD8(+) T cell tolerance are more complex than the proposed "signal 1 in the absence of signal 2" hypothesis. Further exploration of these mechanisms will have direct impact on the design of effective immunotherapy for autoimmune diseases, chronic infections and cancers.

Authors
Huang, X; Yang, Y
MLA Citation
Huang, X, and Yang, Y. "Transient gain of effector function by CD8+ T cells undergoing peripheral tolerance to high-dose self-antigen." Eur J Immunol 34.5 (May 2004): 1351-1360.
PMID
15114668
Source
pubmed
Published In
European Journal of Immunology
Volume
34
Issue
5
Publish Date
2004
Start Page
1351
End Page
1360
DOI
10.1002/eji.200324734

Persistent Toll-like receptor signals are required for reversal of regulatory T cell-mediated CD8 tolerance.

One chief barrier to cancer immunotherapy is tumor-specific T cell tolerance. Here we compared the ability of hemagglutinin (HA)-encoding recombinant viruses versus 'HA-loaded' dendritic cells to reverse HA-specific CD8 tolerance and to protect mice from tumor challenge. Both vaccines were comparable in activating naive HA-specific CD8(+) T cells. However, in circumstances of established tolerance, viral vaccines could break CD8 tolerance in the presence of CD4(+)CD25(+) regulatory T cells, whereas dendritic cell-based vaccines achieved this only after removal of regulatory T cells or the coadministration of a Toll-like receptor (TLR) ligand or irrelevant virus. These results demonstrate that virus provides TLR signals required for bypassing regulatory T cell-mediated tolerance and emphasize the importance of persistent TLR signals for immunotherapy in the setting of established tolerance.

Authors
Yang, Y; Huang, C-T; Huang, X; Pardoll, DM
MLA Citation
Yang, Y, Huang, C-T, Huang, X, and Pardoll, DM. "Persistent Toll-like receptor signals are required for reversal of regulatory T cell-mediated CD8 tolerance." Nat Immunol 5.5 (May 2004): 508-515.
PMID
15064759
Source
pubmed
Published In
Nature Immunology
Volume
5
Issue
5
Publish Date
2004
Start Page
508
End Page
515
DOI
10.1038/ni1059

Immunotherapy of surgical malignancies.

Authors
Morse, MA; Lyerly, HK; Clay, TM; Abdel-Wahab, O; Chui, SY; Garst, J; Gollob, J; Grossi, PM; Kalady, M; Mosca, PJ; Onaitis, M; Sampson, JH; Seigler, HF; Toloza, EM; Tyler, D; Vieweg, J; Yang, Y
MLA Citation
Morse, MA, Lyerly, HK, Clay, TM, Abdel-Wahab, O, Chui, SY, Garst, J, Gollob, J, Grossi, PM, Kalady, M, Mosca, PJ, Onaitis, M, Sampson, JH, Seigler, HF, Toloza, EM, Tyler, D, Vieweg, J, and Yang, Y. "Immunotherapy of surgical malignancies." Curr Probl Surg 41.1 (January 2004): 15-132. (Review)
PMID
14749625
Source
pubmed
Published In
Current Problems in Surgery
Volume
41
Issue
1
Publish Date
2004
Start Page
15
End Page
132
DOI
10.1016/S0011384003001321

How does the immune system attack cancer?

Authors
Morse, MA; Lyerly, HK; Clay, TM; Abdel-Wahab, O; Chui, SY; Garst, J; Gollob, J; Grossi, PM; Kalady, M; Mosca, PJ; Onaitis, M; Sampson, JH; Seigler, HF; Toloza, EM; Tyler, D; Vieweg, J; Yang, Y
MLA Citation
Morse, MA, Lyerly, HK, Clay, TM, Abdel-Wahab, O, Chui, SY, Garst, J, Gollob, J, Grossi, PM, Kalady, M, Mosca, PJ, Onaitis, M, Sampson, JH, Seigler, HF, Toloza, EM, Tyler, D, Vieweg, J, and Yang, Y. "How does the immune system attack cancer?." Current Problems in Surgery 41.1 (2004): 15-132.
Source
scival
Published In
Current Problems in Surgery
Volume
41
Issue
1
Publish Date
2004
Start Page
15
End Page
132
DOI
10.1016/j.cpsurg.2003.08.001

Warfarin-induced skin necrosis in a patient with a mutation of the prothrombin gene [3]

Authors
Yang, Y; Algazy, KM
MLA Citation
Yang, Y, and Algazy, KM. "Warfarin-induced skin necrosis in a patient with a mutation of the prothrombin gene [3]." New England Journal of Medicine 340.9 (1999): 735--.
PMID
10068331
Source
scival
Published In
The New England journal of medicine
Volume
340
Issue
9
Publish Date
1999
Start Page
735-
DOI
10.1056/NEJM199903043400912

Transduction of dendritic cells by DNA viral vectors directs the immune response to transgene products in muscle fibers

Immune responses to vector-corrected cells have limited the application of gene therapy for treatment of chronic disorders such as inherited deficiency states. We have found that recombinant adeno-associated virus (AAV) efficiently transduces muscle fibers in vivo without activation of cellular and humoral immunity to neoantigenic transgene products such as β- galactosidase, which differs from the experience with recombinant adenovirus, where vibrant T-cell responses to the transgene product destroy the targeted muscle fibers. T cells activated following intramuscular administration of adenovirus expressing lacZ (AdlacZ) can destroy AAVlacZ- transduced muscle fibers, indicating a prior state of immunologic nonresponsiveness in the context of AAV gene therapy. Adoptive transfer of dendritic cells infected with AdlacZ leads to immune mediated elimination of AAVlacZ-transduced muscle fibers. AAVlacZ-transduced antigen-presenting cells fail to demonstrate β-galactosidase activity and are unable to elicit transgene immunity in adoptive transfer experiments. These studies indicate that vector-mediated transduction of dendritic cells is necessary for cellular immune responses to muscle gene therapy, a step which AAV avoids, providing a useful biological niche for its use in gene therapy.

Authors
Jooss, K; Yang, Y; Fisher, KJ; Wilson, JM
MLA Citation
Jooss, K, Yang, Y, Fisher, KJ, and Wilson, JM. "Transduction of dendritic cells by DNA viral vectors directs the immune response to transgene products in muscle fibers." Journal of Virology 72.5 (1998): 4212-4223.
PMID
9557710
Source
scival
Published In
Journal of virology
Volume
72
Issue
5
Publish Date
1998
Start Page
4212
End Page
4223

Amelioration of collagen-induced arthritis by Fas-ligand gene transfer

Both rheumatoid arthritis and animal models of autoimmune arthritis are characterized by hyper-activation of synovial cells and hyperplasia of the synovial membrane. The death factor Fas and its ligand (FasL) play pivotal roles in maintaining self tolerance and immune privilege. In both rheumatoid arthritis and animal models of autoimmune arthritis, high levels of Fas are expressed on activated synovial cells and infiltrating leukocytes in the inflamed joints. However, unlike Fas, the levels of FasL expressed in the arthritic joints are extremely low, and most activated synovial cells survive despite high levels of Fas expression. To upregulate FasL expression in the arthritic joints, we have generated a recombinant replication-defective adenovirus carrying FasL gene: injection of the FasL virus into inflamed joints conferred high levels of FasL expression, induced apoptosis of synovial cells and ameliorated collagen-induced arthritis in DBA/1 mice. The Fas-ligand virus also inhibited the production of interferon-g by collagen-specific T cells. Coadministration of Fas-immunoglobulin fusion protein with the Fas-ligand virus prevented these effects, demonstrating the specificity of the Fas-ligand virus. Thus, FasL gene transfer at the site of inflammation effectively ameliorates autoimmune disease.

Authors
Zhang, H; Yang, Y; Horton, JL; Samoilova, EB; Judge, TA; Turka, LA; Wilson, JM; Chen, Y
MLA Citation
Zhang, H, Yang, Y, Horton, JL, Samoilova, EB, Judge, TA, Turka, LA, Wilson, JM, and Chen, Y. "Amelioration of collagen-induced arthritis by Fas-ligand gene transfer." FASEB Journal 12.4 (1998): A309-.
Source
scival
Published In
The FASEB journal : official publication of the Federation of American Societies for Experimental Biology
Volume
12
Issue
4
Publish Date
1998
Start Page
A309

Amelioration of collagen-induced arthritis by CD95 (Apo-1/Fas)-ligand gene transfer

Both rheumatoid arthritis and animal models of autoimmune arthritis are characterized by hyperactivation of synovial cells and hyperplasia of the synovial membrane. The activated synovial cells produce inflammatory cytokines and degradative enzymes that lead to destruction of cartilage and bones. Effective treatment of arthritis may require elimination of most or all activated synovial cells. The death factor Fas/Apo-1 and its ligand (FasL) play pivotal roles in maintaining self-tolerance and immune privilege. Fas is expressed constitutively in most tissues, and is dramatically upregulated at the site of inflammation. In both rheumatoid arthritis and animal models of autoimmune arthritis, high levels of Fas are expressed on activated synovial cells and infiltrating leukocytes in the inflamed joints. Unlike Fas, however, the levels of FasL expressed in the arthritic joints are extremely low, and most activated synovial cells survive despite high levels of Fas expression. To upregulate FasL expression in the arthritic joints, we have generated a recombinant replication-defective adenovirus carrying FasL gene; injection of the FasL virus into inflamed joints conferred high levels of FasL expression, induced apoptosis of synovial cells, and ameliorated collagen-induced arthritis in DBA/1 mice. The Fas-ligand virus also inhibited production of interferon-γ by collagen-specific T cells. Coadministration of Fas-immunoglobulin fusion protein with the Fas-ligand virus prevented these effects, demonstrating the specificity of the Fas-ligand virus. Thus, FasL gene transfer at the site of inflammation effectively ameliorates autoimmune disease.

Authors
Zhang, H; Yang, Y; Horton, JL; Samoilova, EB; Judge, TA; Turka, LA; Wilson, JM; Chen, Y
MLA Citation
Zhang, H, Yang, Y, Horton, JL, Samoilova, EB, Judge, TA, Turka, LA, Wilson, JM, and Chen, Y. "Amelioration of collagen-induced arthritis by CD95 (Apo-1/Fas)-ligand gene transfer." Journal of Clinical Investigation 100.8 (1997): 1951-1957.
PMID
9329958
Source
scival
Published In
Journal of Clinical Investigation
Volume
100
Issue
8
Publish Date
1997
Start Page
1951
End Page
1957
DOI
10.1172/JCI119726

Recombinant adeno-associated virus for muscle directed gene therapy

Although gene transfer with adeno-associated virus (AAV) vectors has typically been low, transduction can be enhanced in the presence of adenovirus gene products through the formation of double stranded, non-integrated AAV genomes. We describe the unexpected finding of high level and stable transgene expression in mice following intramuscular injection of purified recombinant AAV (rAAV). The rAAV genome is efficiently incorporated into nuclei of differentiated muscle fibers where it persists as head-to-tail concatamers. Fluorescent in situ hybridization of muscle tissue suggests single integration sites. Neutralizing antibody against AAV capsid proteins does not prevent readministration of vector. Remarkably, no humoral or cellular immune responses are elicited to the neoantigenic transgene product E. coli β-galactosidase. The favorable biology of rAAV in muscle-directed gene therapy described in this study expands the potential of this vector for the treatment of inherited and acquired diseases.

Authors
Fisher, KJ; Jooss, K; Alston, J; Yang, Y; Haecker, SE; High, K; Pathak, R; Raper, SE; Wilson, JM
MLA Citation
Fisher, KJ, Jooss, K, Alston, J, Yang, Y, Haecker, SE, High, K, Pathak, R, Raper, SE, and Wilson, JM. "Recombinant adeno-associated virus for muscle directed gene therapy." Nature Medicine 3.3 (1997): 306-312.
PMID
9055858
Source
scival
Published In
Nature Medicine
Volume
3
Issue
3
Publish Date
1997
Start Page
306
End Page
312
DOI
10.1038/nm0397-306

Immunology of gene therapy with adenoviral vectors in mouse skeletal muscle

Skeletal muscle is an attractive target for somatic gene transfer of both acquired and inherited disorders. Direct injection of adenoviral vectors in the skeletal muscle leads to recombinant gene expression in a large number of muscle fibers. Transgene expression has been transient in most organs and associated with substantial inflammation when experiments are performed in adult immune competent mice. In this report, we utilize a variety of in vivo and in vitro models of T and B cell function to characterize the nature of the Immune response to adenoviral vectors injected into murine skeletal muscle. Cellular immunity dependent on CD4+ and CD8+ T cells contributes to the loss of recombinant gene expression and the development of localized inflammation. Antigen specific activation of T cells occurs to both viral proteins and the reporter gene β-galactosidase. Systemic levels of neutralizing antibody to the capsid proteins of the vector are also generated. Destructive immune responses responsible for loss of transgene expression are largely directed against β-galactosidase in that transgene expression was stable when β-galactosidase was eliminated as a neoantigen in mice transgenic for lacZ. A strategy to prevent the cellular and humoral immunity to this therapy was developed based on transiently ablating CD4+ T cell activation at the time of vector delivery. Encouraging results were obtained when vector was administered with one of several immune modulating agents including cyclophosphamide, mAb to CD4+ cells, and mAb to CD40 ligand. These studies indicate that cellular and humoral immune responses are elicited in the context of gene therapy directed to skeletal muscle with adenoviral vectors. Transient ablation of CD4+ T cell activation prevents the effector responses of the CD8+ T and B cells.

Authors
Yang, Y; Haecker, SE; Su, Q; Wilson, JM
MLA Citation
Yang, Y, Haecker, SE, Su, Q, and Wilson, JM. "Immunology of gene therapy with adenoviral vectors in mouse skeletal muscle." Human Molecular Genetics 5.11 (1996): 1703-1712.
PMID
8922997
Source
scival
Published In
Human Molecular Genetics
Volume
5
Issue
11
Publish Date
1996
Start Page
1703
End Page
1712
DOI
10.1093/hmg/5.11.1703

Biology of adenovirus vectors with E1 and E4 deletions for liver- directed gene therapy

Recombinant adenoviruses with E1 sequences deleted efficiently transfer genes into a wide variety of target cells. Antigen- and nonantigen-specific responses to the therapy lead to toxicity, loss of transgene expression, and difficulties with vector readministration. We have created new cell lines that allowed the isolation of more disabled adenovirus vectors that have both El and E4 deletions. Studies with murine models of liver-directed gene therapy indicated that the E1- and E4-deleted vector expresses fewer virus proteins and induces less apoptosis, leading to blunted host responses and an improved safety profile. The impact of the E4 deletion on the stability of vector expression was confounded by immune responses to the transgene product, which in this study was β-galactosidase. When transgene responses were eliminated, the doubly deleted vector was substantially more stable in mouse liver than was the E1-deleted construct. These studies indicate that adenovirus vectors with both E1 and E4 deletions may have advantages in terms of safety and efficacy over first-generation constructs for liver-directed gene therapy.

Authors
Gao, G-P; Yang, Y; Wilson, JM
MLA Citation
Gao, G-P, Yang, Y, and Wilson, JM. "Biology of adenovirus vectors with E1 and E4 deletions for liver- directed gene therapy." Journal of Virology 70.12 (1996): 8934-8943.
PMID
8971023
Source
scival
Published In
Journal of virology
Volume
70
Issue
12
Publish Date
1996
Start Page
8934
End Page
8943

CD40 ligand-dependent T cell activation: Requirement of B7-CD28 signaling through CD40

The role of CD40 ligand (CD40L) in the primary activation of T cells is not clear. The cellular and humoral immune responses to adenoviral vectors in a murine model of liver-directed gene transfer were studied to define the mechanisms responsible for CD40L-dependent T cell priming. CD40L-deficient mice did not develop effective cytotoxic T cells to transduced hepatocytes, and T cell-dependent B cell responses were absent. Full reconstitution of cellular and humoral immunity was achieved in CD40L-deficient mice by administration of an activating antibody to CD40 that increased expression of B7.2 on spleen cells. Wild-type mice could be made nonresponsive to vector by administration of antibodies to B7. Thus, CD40L-dependent activation of T cells occurs through signaling of CD40 in the antigen-presenting cell to enhance requisite costimulatory pathways that include B7.

Authors
Yang, Y; Wilson, JM
MLA Citation
Yang, Y, and Wilson, JM. "CD40 ligand-dependent T cell activation: Requirement of B7-CD28 signaling through CD40." Science 273.5283 (1996): 1862-1864.
PMID
8791591
Source
scival
Published In
Science
Volume
273
Issue
5283
Publish Date
1996
Start Page
1862
End Page
1864

Role of viral antigens in destructive cellular immune responses to adenovirus vector-transduced cells in mouse lungs

Adenoviruses missing E1 have been used as gene delivery vectors to the lungs for the treatment of cystic fibrosis. Transient expression of the recombinant gene and the development of inflammation have been two major limitations to the application of first-generation recombinant adenoviruses for gene therapy. Studies with mouse models of liver- and lung-directed gene therapy suggested that CD8+ cytotoxic T lymphocytes (CTLs) are effectors that contribute to extinction of transgene expression. The precise antigens responsible for activation of CTLs have not been identified. In this study, we examine the relative contributions of vital proteins versus the transgene product to the activation of CTLs which eliminate transgene-containing cells in mouse lungs. Instillation of a lacZ-expressing virus into the lungs of C57BL/6 mice elicited CTL responses to both viral proteins and the transgene product, β-galactosidase, which collectively contribute to loss of transgene expression in mouse airways. Similar results were obtained in two experimental models in which the animals should be tolerant to the transgene, i.e., lacZ virus delivered to an animal transgenic for lacZ and a virus expressing the liver-specific enzyme ornithine transcarbamylase administered to the lungs of various strains of immune-competent mice. These data confirm the hypothesis that CTLs specific for vital antigens contribute to the problem of transgene instability in mouse lungs and indicate that CTLs specific for transgene product alone cannot account far the observed problem.

Authors
Yang, Y; Su, Q; Wilson, JM
MLA Citation
Yang, Y, Su, Q, and Wilson, JM. "Role of viral antigens in destructive cellular immune responses to adenovirus vector-transduced cells in mouse lungs." Journal of Virology 70.10 (1996): 7209-7212.
PMID
8794368
Source
scival
Published In
Journal of virology
Volume
70
Issue
10
Publish Date
1996
Start Page
7209
End Page
7212

Transient subversion of CD40 ligand function diminishes immune responses to adenovirus vectors in mouse liver and lung tissues

First-generation adenovirus vectors will have limited application in gene therapy for chronic diseases because of destructive host immune responses. Important immune effectors include CD8+ T cells, which mediate target cell destruction and ablate transgene expression, and B cells, which produce neutralizing antibodies that block effective readministration of vector. Previous studies indicated that activation of CD4+ T cells by virus capsid proteins is necessary for full realization of effector function of CD8+ T cells and B cells. In this paper, we present a strategy for preventing CD4+ T-cell activation by an adenovirus vector delivered to mouse liver and lung tissues which is based on interfering with T-cell priming via CD40 ligand-CD40 interactions. Adenovirus transgene expression was stabilized in mice genetically deficient in CD40 ligand (CD40L), and neutralizing antibody to adenovirus did not develop, allowing efficient readministration of vector. A transient blockade of T-cell activation with an antibody to CD40L infused into the animal at the time of adenovirus vector-mediated gene transfer led to stabilization of transgene expression and diminished production of neutralizing antibody, allowing readministration of vector. In vitro T-cell assays suggested that a block in the primary activation of CD4+ T cells was responsible for the lack of B- cell- and cytotoxic-T-cell-dependent responses. This suggests a strategy for improving the potential of adenovirus vectors based on administration of an antibody to CD40L at the time of vector administration.

Authors
Yang, Y; Su, Q; Grewal, IS; Schilz, R; Flavell, RA; Wilson, JM
MLA Citation
Yang, Y, Su, Q, Grewal, IS, Schilz, R, Flavell, RA, and Wilson, JM. "Transient subversion of CD40 ligand function diminishes immune responses to adenovirus vectors in mouse liver and lung tissues." Journal of Virology 70.9 (1996): 6370-6377.
PMID
8709265
Source
scival
Published In
Journal of virology
Volume
70
Issue
9
Publish Date
1996
Start Page
6370
End Page
6377

Cyclophosphamide diminishes inflammation and prolongs transgene expression following delivery of adenoviral vectors to mouse liver and lung

Immune responses to adenovirus-mediated gene transfer contribute to the problems of transient recombinant gene expression, inflammation, and difficulties with vector readministration. Activation of CD4+ T cells is required for full realization of effector function of both CD8+ T cells (i.e., cytotoxic T cells) and B cells (i.e., neutralizing antibody). We evaluate in this study the effectiveness of a short course of high-dose cyclophosphamide to block immune responses in mice administered vector into lung and liver of C57BL/6 mice. Administration of cyclophosphamide with vector directed to liver blocked activation and mobilization of both CD4+ and CD8+ T cells. As a result, transgene expression was prolonged, inflammation was reduced, and, at the higher doses of cyclophosphamide, formation of neutralizing antibody was prevented and the vector was successfully readministered. Similar studies in the lung demonstrated an effective blockade of T and B cell responses. In contrast to the liver, where it was easier to stabilize transgene expression than to prevent neutralizing antibody, cyclophosphamide prevented the formation of neutralizing antibodies at all doses in the lung, whereas stabilization of transgene expression was only achieved at the highest dose. These experiments begin to define the parameters by which cyclophosphamide could be used as an adjunct in gene therapy.

Authors
Jooss, K; Yang, Y; Wilson, JM
MLA Citation
Jooss, K, Yang, Y, and Wilson, JM. "Cyclophosphamide diminishes inflammation and prolongs transgene expression following delivery of adenoviral vectors to mouse liver and lung." Human Gene Therapy 7.13 (1996): 1555-1566.
PMID
8864756
Source
scival
Published In
Human Gene Therapy
Volume
7
Issue
13
Publish Date
1996
Start Page
1555
End Page
1566

A replication-defective human adenovirus recombinant serves as a highly efficacious vaccine carrier

In this manuscript, an E1 and E3 deleted adenoviral recombinant expressing the rabies virus glycoprotein (G protein) under the control of the cytomegalovirus early promoter was tested for induction of a rabies virus-specific immune response in mice. The construct was found to induce neutralizing antibodies and cytolytic T cells to rabies virus. Mice vaccinated with the adenoviral construct either by the systemic route or by application into the airways were protected against a subsequent infection with a virulent strain of rabies virus. The efficacy of the replication-defective construct was far superior to that of a well-characterized vaccinia rabies glycoprotein recombinant.

Authors
Xiang, ZQ; Yang, Y; Wilson, JM; Ertl, HCJ
MLA Citation
Xiang, ZQ, Yang, Y, Wilson, JM, and Ertl, HCJ. "A replication-defective human adenovirus recombinant serves as a highly efficacious vaccine carrier." Virology 219.1 (1996): 220-227.
PMID
8623532
Source
scival
Published In
Virology
Volume
219
Issue
1
Publish Date
1996
Start Page
220
End Page
227
DOI
10.1006/viro.1996.0239

Transient immune blockade prevents formation of neutralizing antibody to recombinant adenovirus and allows repeated gene transfer to mouse liver

The hepatotropic properties of human adenoviruses have been used to develop vectors for in vivo liver-directed gene therapy. Current limitations of this vector system are the associated hepatitis that develops as a result of antigen-specific cellular immune responses and the difficulty in accomplishing repeated gene transfer. This study uses mouse models to define immune responses of the recipient animal that have previously been shown to prevent successful re-administration of virus and suggests approaches for preventing the development of these blocking immune responses. Our studies are most consistent with class II MHC-dependent activation of T helper cells and B cells to capsid proteins of the input virus leading to the production of antiviral neutralizing antibody following a primary exposure to virus; this capsid-specific antibody appears to bind to virus and prevents entry in the context of a second administration of virus. Transient ablation of CD4 function at the time of virus administration prevents the formation of neutralizing antibody thereby allowing efficient gene transfer after at least two subsequent administrations of virus. Experiments in β2m- mice and C57BL/6 mice treated with IL-12 suggested a more selective ablation of immune function based on inhibiting the activation of the T(H2) subset of T helper cells. From these studies on immune mechanisms it is hoped that Viable strategies can be developed to overcome the problem of humoral immunity that occurs after the initial genetic therapy.

Authors
Yang, Y; Greenough, K; Wilson, JM
MLA Citation
Yang, Y, Greenough, K, and Wilson, JM. "Transient immune blockade prevents formation of neutralizing antibody to recombinant adenovirus and allows repeated gene transfer to mouse liver." Gene Therapy 3.5 (1996): 412-420.
PMID
9156802
Source
scival
Published In
Gene Therapy
Volume
3
Issue
5
Publish Date
1996
Start Page
412
End Page
420

Immune responses to viral antigens versus transgene product in the elimination of recombinant adenovirus-infected hepatocytes in vivo

Human adenoviruses have been developed as an attractive vehicle for in vivo liver-directed gene therapy. Problems with the application of first generation recombinant adenoviruses to liver-directed gene therapy have been transient expression of the recombinant gene and development of hepatitis. Previous studies in mouse models of gene transfer to liver and lung suggested that MHC class I-restricted cytotoxic T lymphocytes (CTLs) to viral antigens may be effectors in the elimination of transgene expression. The goal of this study was to evaluate the importance of viral antigens versus transgene product in inducing CTL mediated hepatocyte destruction in vivo. Immunization of C57BL/6 mice with a lacZ-expressing adenovirus elicited CTL responses to both viral antigens and the transgene product, β-galactosidase (β-gal). Adoptive transfer experiments, as well as studies involving lacZ-transgenic mice (ROSA-26) revealed that CTLs to viral antigens are sufficient to destroy virus-infected hepatocytes, indicating that CTLs to β-gal can not solely account for the observed hepatocyte destruction that has characterized the use of first generation viruses. In addition, we confirm that B cell-mediated events do not participate in destruction of hepatocytes in vivo, despite the production of virus- and β-gal-specific antibodies. These data confirm the hypothesis that viral gene expression elicits host responses that contribute to the problem of transgene instability. Recombinant adenoviruses must be redesigned to diminish viral gene expression if they are to be used in the treatment of chronic diseases.

Authors
Yang, Y; Ku, J; Su, Q; Ertl, HCJ; Wilson, JM
MLA Citation
Yang, Y, Ku, J, Su, Q, Ertl, HCJ, and Wilson, JM. "Immune responses to viral antigens versus transgene product in the elimination of recombinant adenovirus-infected hepatocytes in vivo." Gene Therapy 3.2 (1996): 137-144.
PMID
8867861
Source
scival
Published In
Gene Therapy
Volume
3
Issue
2
Publish Date
1996
Start Page
137
End Page
144

Upregulation of class I major histocompatibility complex antigens by interferon γ is necessary for T-cell-mediated elimination of recombinant adenovirus-infected hepatocytes in vivo

Recombinant adenoviruses are attractive vehicles for liver-directed gene therapy because of the high efficiency with which they transfer genes to hepatocytes in vivo. First generation recombinant adenoviruses deleted of E1 sequences also express recombinant and early and late viral genes, which lead to development of destructive cellular immune responses. Previous studies indicated that class I major histocompatibility complex (MHC)-restricted cytotoxic T lymphocytes (CTLs) play a major role in eliminating virus- infected cells. The present studies utilize mouse models to evaluate the role of T-helper cells in the primary response to adenovirus-mediated gene transfer to the liver. In vivo ablation of CD4+ cells or interferon γ (IFN- γ) was sufficient to prevent the elimination of adenovirus-transduced hepatocytes, despite the induction of a measurable CTL response. Mobilization of an effective T(H1) response as measured by in vitro proliferation assays was associated with substantial upregulation of MHC class I expression, an effect that was prevented in IFN-γ-deficient animals. These results suggest that elimination of virus-infected hepatocytes in a primary exposure to recombinant adenovirus requires both induction of antigen-specific CTLs as well as sensitization of the target cell by T(H1)-mediated activation of MHC class I expression.

Authors
Yang, Y; Xiang, Z; Ertl, HCJ; Wilson, JM
MLA Citation
Yang, Y, Xiang, Z, Ertl, HCJ, and Wilson, JM. "Upregulation of class I major histocompatibility complex antigens by interferon γ is necessary for T-cell-mediated elimination of recombinant adenovirus-infected hepatocytes in vivo." Proceedings of the National Academy of Sciences of the United States of America 92.16 (1995): 7257-7261.
PMID
7638177
Source
scival
Published In
Proceedings of the National Academy of Sciences of USA
Volume
92
Issue
16
Publish Date
1995
Start Page
7257
End Page
7261
DOI
10.1073/pnas.92.16.7257

Clearance of adenovirus-infected hepatocytes by MHC class I-restricted CD4+ CTLs in vivo

E1-deleted recombinant adenoviruses have been developed for liver- directed gene therapy because efficient gene transfer to hepatocytes can be achieved in vivo. However, these viruses also express viral proteins in hepatocytes, leading to the development of destructive immune responses. Our previous studies indicated that MHC class I-restricted CD8+ CTLs are major effectors in eliminating virus-infected cells, and CD4+ cells are also necessary in developing a fully competent CTL response by secretion of IFN- γ, which sensitizes the virus infected hepatocytes to CTLs through up- regulation of MHC class I expression. In this study, we have used adoptive transfer techniques in combination with mice deficient in immune functions to further define the role of CD4+ cells in the primary response to adenovirus- mediated gene transfer to the liver. Our studies indicate that CD4+ cells alone are capable of destroying virus-infected hepatocytes. Adoptive transfer experiments with β2m mice along with in vitro CTL assays suggest that these CD4+ can act as CTL effectors, which are MHC class I-restricted. Depletion of these CD4+ effectors in vivo leads to prolongation of adenovirus-mediated transgene expression in hepatocytes. These results suggest that class I- restricted CD4+ CTLs contribute to elimination of adenovirus-transduced hepatocytes and extend our understanding of functional importance of CD4+ cells in viral pathogenesis.

Authors
Yang, Y; Wilson, JM
MLA Citation
Yang, Y, and Wilson, JM. "Clearance of adenovirus-infected hepatocytes by MHC class I-restricted CD4+ CTLs in vivo." Journal of Immunology 155.5 (1995): 2564-2570.
PMID
7650386
Source
scival
Published In
Journal of Immunology
Volume
155
Issue
5
Publish Date
1995
Start Page
2564
End Page
2570

Recombinant IL-12 prevents formation of blocking IgA antibodies to recombinant adenovirus and allows repeated gene therapy to mouse lung

Enthusiasm for the use of recombinant adenoviruses in gene therapy has been tempered by the problematic immune responses that develop to the virus and virus-infected cells. Humoral immune responses to the input viral proteins generate neutralizing antibodies that thwart attempts to effectively administer the therapy more than once. Previous studies in murine models of gene therapy for cystic fibrosis (CF) have shown that the formation of adenoviral antibodies of the IgA subtype, a process that is dependent on T helper cells of the TH2 subset, contributes to a block in gene transfer that occurs following a second administration of virus. We show in this report that coadministration of interferon-γ (IFN-γ) (or interleukin-12, which activates TH1 cells to secrete IFN-γ) with the recombinant adenovirus into the airway of C57BL/6 mice diminishes the activation of TH2 cells and formation of neutralizing antibody, allowing for efficient readministration of recombinant virus. This suggests a strategy for gene therapy of CF in which administration of a short-acting immune modulator at the time of gene therapy may be sufficient to overcome the problems of humoral immunity.

Authors
Yang, Y; Trinchieri, G; Wilson, JM
MLA Citation
Yang, Y, Trinchieri, G, and Wilson, JM. "Recombinant IL-12 prevents formation of blocking IgA antibodies to recombinant adenovirus and allows repeated gene therapy to mouse lung." Nature Medicine 1.9 (1995): 890-893.
PMID
7585213
Source
scival
Published In
Nature Medicine
Volume
1
Issue
9
Publish Date
1995
Start Page
890
End Page
893

Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses

Recombinant adenoviruses are an attractive vehicle for gene therapy to the lung in the treatment of cystic fibrosis (CF). First-generation viruses deleted of E1a and E1b transduce genes into airway epithelial cells in vivo; however, expression of the transgene is transient and associated with substantial inflammatory responses, and gene transfer is significantly reduced following a second administration of the virus. In this study, we have used mice deficient in immunological effector functions in combination with adoptive and passive transfer techniques to define antigen-specific cellular and humoral immune responses that underlie these important limitations. Our studies indicate that major histocompatibility complex class I-restricted CD8+ cytotoxic T lymphocytes are activated in response to newly synthesized antigens, leading to destruction of virus infected cells and loss of transgene expression. Major histocompatibility complex class II- associated presentation of exogenous viral antigens activates CD4+ T-helper (T(H)) cells of the T(H1) subset and, to a lesser extent, of the T(H2) subset. CD4+ cell-mediated responses are insufficient in the absence of cytotoxic T cells to completely eliminate transgene containing cells; however, they contribute to the formation of neutralizing antibodies in the airway which block subsequent adenovirus-mediated gene transfer. Definition of immunological barriers to gene therapy of cystic fibrosis should facilitate the design of rational strategies to overcome them.

Authors
Yang, Y; Li, Q; Ertl, HCJ; Wilson, JM
MLA Citation
Yang, Y, Li, Q, Ertl, HCJ, and Wilson, JM. "Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses." Journal of Virology 69.4 (1995): 2004-2015.
PMID
7884845
Source
scival
Published In
Journal of Virology
Volume
69
Issue
4
Publish Date
1995
Start Page
2004
End Page
2015

Gene therapy in a xenograft model of cystic fibrosis lung corrects chloride transport more effectively than the sodium defect

We have developed a model of gene therapy for cystic fibrosis (CF) lung disease, based on growth of human CF bronchial xenografts in nu/nu mice. We now report an evaluation of the primary abnormalities in CF lung epithelia- defective CI secretion and Na hyperabsorption-in xenografts following adenovirus-mediated gene transfer. In vivo infection of CF xenografts with a cystic fibrosis transmembrane regulator (CFTR) recombinant adenovirus, at a multiplicity of infection equal to 100, was sufficient to reconstitute near normal levels of cAMP-stimulated CI transport, despite transducing only 5% of cells in the pseudostratified epithelium. Correction in sodium hyperabsorption was partial and variable. These experiments define aspects of adenovirus-mediated gene therapy relevant to CF protocols based on intrapulmonary genetic reconstitution.

Authors
Goldman, MJ; Yang, Y; Wilson, JM
MLA Citation
Goldman, MJ, Yang, Y, and Wilson, JM. "Gene therapy in a xenograft model of cystic fibrosis lung corrects chloride transport more effectively than the sodium defect." Nature Genetics 9.2 (1995): 126-131.
PMID
7719338
Source
scival
Published In
Nature Genetics
Volume
9
Issue
2
Publish Date
1995
Start Page
126
End Page
131

Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy.

An important limitation that has emerged in the use of adenoviruses for gene therapy has been loss of recombinant gene expression that occurs concurrent with the development of pathology in the organ expressing the transgene. We have used liver-directed approaches to gene therapy in mice to study mechanisms that underlie the problems with transient expression and pathology that have characterized in vivo applications of first-generation recombinant adenoviruses (i.e., those deleted of E1a and E1b). Our data are consistent with the following hypothesis. Cells harboring the recombinant viral genome express the transgene as desired; however, low-level expression of viral genes also occurs. A virus-specific cellular immune response is stimulated that leads to destruction of the genetically modified hepatocytes, massive hepatitis, and repopulation of the liver with nontransgene-containing hepatocytes. These findings suggest approaches for improving recombinant adenoviruses that are based on further crippling the virus to limit expression of nondeleted viral genes.

Authors
Yang, Y; Nunes, FA; Berencsi, K; Furth, EE; Gönczöl, E; Wilson, JM
MLA Citation
Yang, Y, Nunes, FA, Berencsi, K, Furth, EE, Gönczöl, E, and Wilson, JM. "Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy." Proc Natl Acad Sci U S A 91.10 (May 10, 1994): 4407-4411.
PMID
8183921
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
91
Issue
10
Publish Date
1994
Start Page
4407
End Page
4411

Ultrastructural localization of variant forms of cystic fibrosis transmembrane conductance regulator in human bronchial epithelial of xenografts.

Cystic fibrosis (CF) is caused by mutations in the gene encoding a cyclic adenosine monophosphate (cAMP)-regulated chloride (CI) channel called the CF transmembrane conductance regulator (CFTR). Previous in vitro studies have indicated that the most common mutation, delta F508 CFTR (a deletion of phenylalanine 508), encodes a protein that is trapped in the endoplasmic reticulum (ER), leading to loss of cAMP-regulated CI transport at the plasma membrane. Another common variant, G551D CFTR (a G-->D missense mutation at position 551), is properly transported to the plasma membrane but is unresponsive to cAMP. These hypotheses are based primarily on studies in culture cells. We have attempted to extend the in vitro experiments by characterizing the molecular pathogenesis of the common mutations, delta F508 and G551D, in the context of a more relevant setting, the pseudostratified epithelium of a proximal human airway. Recombinant adenoviruses were used to transduce normal and variant forms of CFTR into surface epithelial cells of human bronchial xenografts grown in nu/nu mice. Recombinant forms of CFTR RNA and protein were expressed at levels that exceed expression of the endogenous gene. Immunolocalization of CFTR at the light and electron microscopic level indicated that products of the wild type and G551D alleles are found primarily at the apical plasma membrane of ciliated cells, while the delta F508 variant is distributed diffusely throughout the ER. Our data support previous observations primarily made in vitro that the G551D variant is a dysfunctional channel that is properly processed and that the delta F508 variant undergoes biosynthetic arrest at the level of the ER.

Authors
Yang, Y; Engelhardt, JF; Wilson, JM
MLA Citation
Yang, Y, Engelhardt, JF, and Wilson, JM. "Ultrastructural localization of variant forms of cystic fibrosis transmembrane conductance regulator in human bronchial epithelial of xenografts." American journal of respiratory cell and molecular biology 11.1 (1994): 7-15.
PMID
7517144
Source
scival
Published In
American journal of respiratory cell and molecular biology
Volume
11
Issue
1
Publish Date
1994
Start Page
7
End Page
15

Inactivation of E2a in recombinant adenoviruses improves the prospect for gene therapy in cystic fibrosis

Although first generation recombinant adenoviruses, deleted of sequences spanning E1a and E1b, have been useful for in vivo applications of gene therapy, expression of the recombinant gene has been transient and often associated with the development of inflammation. We show that with first generation adenovirus-mediated gene transfer to the mouse lung, viral proteins are expressed leading to destructive cellular immune responses and repopulation of the lung with nontransgene containing cells. Second generation E1 deleted viruses further crippled by a temperature sensitive mutation in the E2a gene were associated with substantially longer recombinant gene expression and less inflammation. Stable expression of human CF transmembrane conductance regulator has been achieved in lungs of CF mice instilled with a second generation virus.

Authors
Yang, Y; Nunes, FA; Berencsi, K; Gönczöl, E; Engelhardt, JF; Wilson, JM
MLA Citation
Yang, Y, Nunes, FA, Berencsi, K, Gönczöl, E, Engelhardt, JF, and Wilson, JM. "Inactivation of E2a in recombinant adenoviruses improves the prospect for gene therapy in cystic fibrosis." Nature Genetics 7.3 (1994): 362-369.
PMID
7522742
Source
scival
Published In
Nature Genetics
Volume
7
Issue
3
Publish Date
1994
Start Page
362
End Page
369
DOI
10.1038/ng0794-362

MHC class I-restricted cytotoxic T lymphocytes to viral antigens destroy hepatocytes in mice infected with E1-deleted recombinant adenoviruses

The use of E1-deleted recombinant adenoviruses in gene therapy has consistently been associated with transient gene expression and inflammation due to immune-based destruction of the infected cells. We have used murine models of adenovirus-mediated gene transfer to liver to investigate these immunologic mechanisms. Adoptive transfer experiments, as well as studies involving genetic knockout mice, confirmed the original hypothesis that cell-mediated immunity induced by E1-deleted adenovirus destroyed transgene-expressing hepatocytes and defined MHC class I-restricted CD8+ cytolytic lymphocytes as the primary immune effectors for hepatocyte destruction. Responses mediated by CD4+ cells per se were insufficient to mediate destruction of hepatocytes in vivo, despite the activation of virus-specific T helper cells of Th1 subsets. A better understanding of the response of the host to in vivo gene therapy is important in evaluating its usefulness in humans.

Authors
Yang, Y; Ertl, HCJ; Wilson, JM
MLA Citation
Yang, Y, Ertl, HCJ, and Wilson, JM. "MHC class I-restricted cytotoxic T lymphocytes to viral antigens destroy hepatocytes in mice infected with E1-deleted recombinant adenoviruses." Immunity 1.5 (1994): 433-442.
PMID
7533647
Source
scival
Published In
Immunity
Volume
1
Issue
5
Publish Date
1994
Start Page
433
End Page
442

Gene therapy for cystic fibrosis using el-deleted adenovirus: A phase I trial in the nasal cavity the University of North Carolina at Chapel Hill

Authors
Boucher, RC; Knowles, MR; Johnson, LG; Olsen, JC; Pickles, R; Wilson, JM; Engelhardt, J; Yang, Y; Grossman, M
MLA Citation
Boucher, RC, Knowles, MR, Johnson, LG, Olsen, JC, Pickles, R, Wilson, JM, Engelhardt, J, Yang, Y, and Grossman, M. "Gene therapy for cystic fibrosis using el-deleted adenovirus: A phase I trial in the nasal cavity the University of North Carolina at Chapel Hill." Human Gene Therapy 5.5 (1994): 615-639.
PMID
7519885
Source
scival
Published In
Human Gene Therapy
Volume
5
Issue
5
Publish Date
1994
Start Page
615
End Page
639

Gene therapy of cystic fibrosis lung disease using E1 deleted adenoviruses: A phase I trial

Cystic fibrosis (CF) is an autosomal recessive disorder caused by defective ion transport across various epithelia. Multiple organ systems are affected in this disease; however, the pulmonary complications are the most morbid and life limiting. The primary defect in the lung appears to be abnormal mucociliary clearance. Isolation of the gene responsible for CF in 1989 provided impetus for the development of new therapies based on gene therapy. We propose in this protocol a phase I trial to assess the safety and feasibility of treating CF pulmonary disease by directly delivering CFTR-expressing, replication defective adenoviruses to the airway. The rationale for this human protocol was based on extensive preclinical studies in a variety of animal models including human CF airway xenografts and nonhuman primates. In this protocol, 20 CF patients will be treated with CFTR-expressing virus and followed for a) evidence of CFTR gene transfer and expression, b) immunological responses to CFTR or adenoviral proteins, and c) toxicity. Adult CF patients with advanced disease who are considered acceptable candidates for bronchoscopy will be considered. A suspension of virus will be delivered to an isolated segment of the lung via a bronchoscope. Pulmonary samples will be harvested for analyses by follow-up bronchoscopies 4 days, 6 weeks, and 3 months following administration of the virus.

Authors
Wilson, JM; Engelhardt, JF; Grossman, M; Simon, RH; Yang, Y
MLA Citation
Wilson, JM, Engelhardt, JF, Grossman, M, Simon, RH, and Yang, Y. "Gene therapy of cystic fibrosis lung disease using E1 deleted adenoviruses: A phase I trial." Human Gene Therapy 5.4 (1994): 501-519.
PMID
7519452
Source
scival
Published In
Human Gene Therapy
Volume
5
Issue
4
Publish Date
1994
Start Page
501
End Page
519

The common variant of cystic fibrosis transmembrane conductance regulator is recognized by hsp70 and degraded in a pre-Golgi nonlysosomal compartment.

The most common cause of cystic fibrosis is deletion of Phe-508 (delta F508) from the cystic fibrosis transmembrane conductance regulator (CFTR). Previous studies have suggested that delta F508 CFTR is an unstable protein that retains a pattern of glycosylation specific to the endoplasmic reticulum. This report examines the mechanism responsible for the mislocalization of delta F508 CFTR in a human cystic fibrosis epithelial cell line overexpressing recombinant CFTR by virtue of adenovirus-mediated gene transfer. Immunoelectron microscopy confirmed that wild-type CFTR is delivered to the plasma membrane of these cells and that delta F508 CFTR is retained in the endoplasmic reticulum. Pulse-chase studies showed that newly synthesized CFTR complexes with the chaperone hsp70. The wild-type protein dissociates from hsp70 before its transport to the Golgi, and the protein is subsequently degraded in lysosomes. By contrast, the complex formed between delta F508 CFTR and hsp70 is retained in the endoplasmic reticulum and delta F508 CFTR is rapidly degraded in a pre-Golgi nonlysosomal compartment. Thus, hsp70 discriminates between the normal form of CFTR and the form of the protein that most commonly causes cystic fibrosis (delta F508). These findings clarify the mechanism by which mutation causing delta F508 affects the intracellular trafficking of CFTR and suggest another function for hsp70 in ensuring quality control during the biosynthesis of plasma-membrane proteins.

Authors
Yang, Y; Janich, S; Cohn, JA; Wilson, JM
MLA Citation
Yang, Y, Janich, S, Cohn, JA, and Wilson, JM. "The common variant of cystic fibrosis transmembrane conductance regulator is recognized by hsp70 and degraded in a pre-Golgi nonlysosomal compartment." Proc Natl Acad Sci U S A 90.20 (October 15, 1993): 9480-9484.
PMID
7692448
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
90
Issue
20
Publish Date
1993
Start Page
9480
End Page
9484

Molecular basis of defective anion transport in L cells expressing recombinant forms of CFTR.

Cystic fibrosis (CF) is caused by mutations in the gene encoding a chloride channel called the CF transmembrane conductance regulator (CFTR). A single mutation in this gene, deletion of three nucleotides that leads to the absence of phenylalanine 508 (i.e., delta F508), is found on 70% of all CF chromosomes. To explore the molecular mechanism(s) responsible for defective chloride transport in patients with CF, we have studied the processing, localization, and function of wild type (W.T.), delta F508 and G551D CFTR (a G-->D missense mutation at position 551) in retrovirus transduced L cells. Cell transduced with W.T. CFTR expressed a 170 kd CFTR protein that was endoglycosidase H (Endo H) resistant, localized to the plasma membrane, and generated a cAMP-mediated anion conductance (GCl) when stimulated with standard concentrations of forskolin (5 microM), cpt cAMP (400 microM) and IBMX (100 microM). The G551D CFTR was indistinguishable from W.T. CFTR with respect to post-translational processing and localization, but it did not produce a cAMP-activated GCl in response to the standard stimulation cocktail. However, raising the IBMX concentration to 4 mM produced GCl in G551D expressing cells. Cells transduced with delta F508 CFTR expressed an Endo H sensitive CFTR protein (approximately 140 kd) that was found in a cytosolic, perinuclear location. These cells did not respond to the standard cocktail, but approximately 20% of cells increased GCl when the cocktail contained 4 mM IBMX.(ABSTRACT TRUNCATED AT 250 WORDS)

Authors
Yang, Y; Devor, DC; Engelhardt, JF; Ernst, SA; Strong, TV; Collins, FS; Cohn, JA; Frizzell, RA; Wilson, JM
MLA Citation
Yang, Y, Devor, DC, Engelhardt, JF, Ernst, SA, Strong, TV, Collins, FS, Cohn, JA, Frizzell, RA, and Wilson, JM. "Molecular basis of defective anion transport in L cells expressing recombinant forms of CFTR." Hum Mol Genet 2.8 (August 1993): 1253-1261.
PMID
7691345
Source
pubmed
Published In
Human Molecular Genetics
Volume
2
Issue
8
Publish Date
1993
Start Page
1253
End Page
1261

An approach for treating the hepatobiliary disease of cystic fibrosis by somatic gene transfer.

Cystic fibrosis (CF) is an inherited disease of epithelial cell ion transport that is associated with pathology in multiple organ systems, including lung, pancreas, and liver. As treatment of the pulmonary manifestations of CF has improved, management of CF liver disease has become increasingly important in adult patients. This report describes an approach for treating CF liver disease by somatic gene transfer. In situ hybridization and immunocytochemistry analysis of rat liver sections indicated that the endogenous CFTR (cystic fibrosis transmembrane conductance regulator) gene is primarily expressed in the intrahepatic biliary epithelial cells. To specifically target recombinant genes to the biliary epithelium in vivo, recombinant adenoviruses expressing lacZ or human CFTR were infused retrograde into the biliary tract through the common bile duct. Conditions were established for achieving recombinant gene expression in virtually all cells of the intrahepatic bile ducts in vivo. Expression persisted in the smaller bile ducts for the duration of the experiment, which was 21 days. These studies suggest that it may be feasible to prevent CF liver disease by genetically reconstituting CFTR expression in the biliary tract, using an approach that is clinically feasible.

Authors
Yang, Y; Raper, SE; Cohn, JA; Engelhardt, JF; Wilson, JM
MLA Citation
Yang, Y, Raper, SE, Cohn, JA, Engelhardt, JF, and Wilson, JM. "An approach for treating the hepatobiliary disease of cystic fibrosis by somatic gene transfer." Proc Natl Acad Sci U S A 90.10 (May 15, 1993): 4601-4605.
PMID
7685107
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
90
Issue
10
Publish Date
1993
Start Page
4601
End Page
4605

Expression of an abundant alternatively spliced form of the cystic fibrosis transmembrane conductance regulator (CFTR) gene is not associated with a cAMP-activated chloride conductance.

The cystic fibrosis transmembrane conductance regulator (CFTR) gene encodes a cAMP-activated chloride (Cl-) channel, and expression of the full length gene in vitro is sufficient to correct the Cl- conductance defect that is characteristic of cystic fibrosis (CF) epithelial cells. Alternatively spliced forms of CFTR mRNA have been identified in several tissues from normal individuals. One of the alternative transcripts, often present at high levels, results in the in-frame deletion of exon 9. Translation of this transcript would result in a CFTR protein missing the amino terminal portion of the first nucleotide binding fold (NBF). To evaluate the possible function of this form of CFTR, a cDNA representing this transcript (CFTR delta 9) was transduced into CFPAC cells, which are derived from a CF patient. CFTR delta 9 RNA was expressed in the transduced cell lines, but only immature, incompletely glycosylated protein was detectable by Western blot analysis. No increase in cAMP-activated anion permeability was detectable by 125I efflux assay or by means of the halide sensitive dye 6-methoxy-N-(3-sulfopropyl) quinolinium (SPQ). In a second assay system, in vitro synthesized mRNA representing CFTR delta D9 was injected into Xenopus oocytes, but expression of this alternatively spliced form of CFTR was not associated with the appearance of Cl- conductance. These results suggest that the protein produced by the CFTR delta 9 transcript is not properly processed and is not capable of generating Cl- conductance in response to cAMP. Whether this alternative transcript has some other function or represents 'noise' in the mRNA splicing mechanism remains unresolved.

Authors
Strong, TV; Wilkinson, DJ; Mansoura, MK; Devor, DC; Henze, K; Yang, Y; Wilson, JM; Cohn, JA; Dawson, DC; Frizzell, RA
MLA Citation
Strong, TV, Wilkinson, DJ, Mansoura, MK, Devor, DC, Henze, K, Yang, Y, Wilson, JM, Cohn, JA, Dawson, DC, and Frizzell, RA. "Expression of an abundant alternatively spliced form of the cystic fibrosis transmembrane conductance regulator (CFTR) gene is not associated with a cAMP-activated chloride conductance." Hum Mol Genet 2.3 (March 1993): 225-230.
PMID
7684641
Source
pubmed
Published In
Human Molecular Genetics
Volume
2
Issue
3
Publish Date
1993
Start Page
225
End Page
230

Adenovirus-mediated transfer of the CFTR gene to lung of nonhuman primates: Biological efficacy study

We have evaluated the biological efficacy of E1-deleted adenoviruses in baboons for lung-directed gene therapy of cystic fibrosis (CF). The experimental design attempted to simulate a phase I clinical trial with animals receiving a single dose of virus to an isolated pulmonary segment. A total of 14 animals divided into four groups, each of which received escalating doses of virus, were used. Individual animals were necropsied 4 and 21 days after gene transfer and tissues were carefully surveyed for gene expression. Expression of the transgene was localized primarily to the area into which it was infused; the efficiency of recombinant gene expression and the abundance of transgene sequences were proportional to dose and both diminished with time. Transgene expression was found predominantly in alveolar cells with patches of expression in the proximal and distal airway. Analysis of adenoviral protein expression within transgene-expressing cells revealed infrequent expression of the E2a gene and no detectable expression of late genes (i.e., fiber protein). These results suggest that recombinant adenovirus can be used to transfer genes efficiently to the lung of nonhuman primates and that therapeutic strategies of cystic fibrosis may require repetitive administration with current vectors.

Authors
Engelhardt, JF; Simon, RH; Yang, Y; Zepeda, M; Weber-Pendleton, S; Doranz, B; Grossman, M; Wilson, JM
MLA Citation
Engelhardt, JF, Simon, RH, Yang, Y, Zepeda, M, Weber-Pendleton, S, Doranz, B, Grossman, M, and Wilson, JM. "Adenovirus-mediated transfer of the CFTR gene to lung of nonhuman primates: Biological efficacy study." Human Gene Therapy 4.6 (1993): 759-769.
PMID
7514445
Source
scival
Published In
Human Gene Therapy
Volume
4
Issue
6
Publish Date
1993
Start Page
759
End Page
769

Adenovirus-mediated transfer of the CFTR gene to lung of nonhuman primates: Toxicity study

In preparation for human trials of gene therapy for cystic fibrosis (CF), we performed a preclinical study of gene transfer into the lungs of baboons. Recombinant adenovirus vectors containing expression cassettes for human cystic fibrosis transmembrane conductance regulator (CFTR) and Escherichia coli β-galactosidase (lacZ) were instilled through a bronchoscope into limited regions of lung in 14 baboons. A detailed accounting of the extent, distribution, and duration of gene expression is contained in a companion article. In this article, we report the results of toxicity studies in which clinical laboratory tests, chest radiographs, and necropsy studies were used to detect adverse effects. The only adverse effect noted was a mononuclear cell inflammatory response within the alveolar compartment of animals receiving doses of virus that were required to induce detectable gene expression. Minimal inflammation was seen at 107 and 108 pfu/ml, but at 109 and more prominently at 1010 pfu/ml, a perivascular lymphocytic and histocytic infiltrate was seen. The intensity of inflammation increased between 4 and 21 days. At its greatest intensity, there was diffuse alveolar wall damage with intra-alveolar edema. Airways were relatively spared, despite the intensity of alveolar inflammation. Clinical tests did not accurately reflect the presence of lung inflammation, with the exception of chest radiographs which revealed alveolar infiltrates, but only in regions of lung having the greatest intensity inflammation. We conclude that adenovirus-mediated gene transfer into the lungs of baboons is associated with development of alveolar inflammation at high doses of virus.

Authors
Simon, RH; Engelhardt, JF; Yang, Y; Zepeda, M; Weber-Pendleton, S; Grossman, M; Wilson, JM
MLA Citation
Simon, RH, Engelhardt, JF, Yang, Y, Zepeda, M, Weber-Pendleton, S, Grossman, M, and Wilson, JM. "Adenovirus-mediated transfer of the CFTR gene to lung of nonhuman primates: Toxicity study." Human Gene Therapy 4.6 (1993): 771-780.
PMID
7514446
Source
scival
Published In
Human Gene Therapy
Volume
4
Issue
6
Publish Date
1993
Start Page
771
End Page
780

Direct gene transfer of human CFTR into human bronchial epithelia of xenografts with E1-deleted adenoviruses

We describe the use of a human bronchial xenograft model for studying the efficiency and biology of in vivo gene transfer into human bronchial epithelia with recombinant E1 deleted adenoviruses. All cell types in the surface epithelium except basal cells efficiently expressed the adenoviral transduced recombinant genes, lacZ and CFTR, for 3-5 weeks. Stable transgene expression was associated with high level expression of the early adenoviral gene, E2a, in a subset of transgene expressing cells and virtually undetectable expression of the late adenoviral genes encoding the structural proteins, hexon and fiber. These studies begin to address important issues that relate to safety and in vivo efficacy of recombinant adenoviruses for gene delivery into the human airway.

Authors
Engelhardt, JF; Yang, Y; Stratford-Perricaudet, LD; Allen, ED; Kozarsky, K; Perricaudet, M; Yankaskas, JR; Wilson, JM
MLA Citation
Engelhardt, JF, Yang, Y, Stratford-Perricaudet, LD, Allen, ED, Kozarsky, K, Perricaudet, M, Yankaskas, JR, and Wilson, JM. "Direct gene transfer of human CFTR into human bronchial epithelia of xenografts with E1-deleted adenoviruses." Nature Genetics 4.1 (1993): 27-34.
PMID
7685651
Source
scival
Published In
Nature Genetics
Volume
4
Issue
1
Publish Date
1993
Start Page
27
End Page
34
DOI
10.1038/ng0593-27

Submucosal glands are the predominant site of CFTR expression in the human bronchus.

We have used in situ hybridization and immunocytochemistry to characterize the cellular distribution of cystic fibrosis (CF) gene expression in human bronchus. The cystic fibrosis transmembrane conductance regular (CFTR) was primarily localized to cells of submucosal glands in bronchial tissues from non-CF individuals notably in the serous component of the secretory tubules as well as a subpopulation of cells in ducts. Normal distribution of CFTR mRNA was found in CF tissues while expression of CFTR protein was genotype specific, with delta F508 homozygotes demonstrating no detectable protein and compound heterozygotes expressing decreased levels of normally distributed protein. Our data suggest mechanisms whereby defects in CFTR expression could lead to abnormal production of mucus in human lung.

Authors
Engelhardt, JF; Yankaskas, JR; Ernst, SA; Yang, Y; Marino, CR; Boucher, RC; Cohn, JA; Wilson, JM
MLA Citation
Engelhardt, JF, Yankaskas, JR, Ernst, SA, Yang, Y, Marino, CR, Boucher, RC, Cohn, JA, and Wilson, JM. "Submucosal glands are the predominant site of CFTR expression in the human bronchus." Nat Genet 2.3 (November 1992): 240-248.
PMID
1285365
Source
pubmed
Published In
Nature Genetics
Volume
2
Issue
3
Publish Date
1992
Start Page
240
End Page
248
DOI
10.1038/ng1192-240
Show More

Research Areas:

  • Acute Disease
  • Adaptive Immunity
  • Adenoviridae
  • Adenoviridae Infections
  • Adenovirus E1A Proteins
  • Adenovirus E1B Proteins
  • Adenoviruses, Human
  • Adjuvants, Immunologic
  • Adoptive Transfer
  • Aged
  • Alternative Splicing
  • Antibody Formation
  • Antigen Presentation
  • Antigens, CD4
  • Antigens, CD8
  • Antigens, Neoplasm
  • Antigens, Viral
  • Antineoplastic Agents
  • Autoantigens
  • Autoimmune Diseases
  • Autoimmunity
  • Blotting, Western
  • CD4-Positive T-Lymphocytes
  • CD8-Positive T-Lymphocytes
  • Cell Proliferation
  • Chaperonins
  • Chloride Channels
  • Coculture Techniques
  • Combined Modality Therapy
  • Cyclic AMP
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Cytokines
  • Cytotoxicity, Immunologic
  • DNA, Viral
  • Dendritic Cells
  • Dependovirus
  • Disease Models, Animal
  • Electric Conductivity
  • Endoplasmic Reticulum
  • Endosomes
  • Extracellular Signal-Regulated MAP Kinases
  • Female
  • Flow Cytometry
  • Gene Deletion
  • Gene Knock-In Techniques
  • Gene Transfer Techniques
  • Gene therapy
  • Genes, Bacterial
  • Genes, Viral
  • Genetic Therapy
  • Germinal Center
  • Glucose
  • Graft vs Host Disease
  • Growth Inhibitors
  • HLA Antigens
  • HLA-C Antigens
  • Heat-Shock Proteins
  • Hemagglutinins
  • Hematologic Neoplasms
  • Hematopoietic Stem Cell Transplantation
  • Heparitin Sulfate
  • Histocompatibility
  • Histocompatibility Testing
  • Humans
  • Immune System
  • Immune Tolerance
  • Immunity, Cellular
  • Immunity, Innate
  • Immunologic Memory
  • Immunosuppressive Agents
  • Immunotherapy
  • Influenza A virus
  • Interferon Type I
  • Interferon-beta
  • Interleukin-10
  • Interleukin-12
  • Interleukin-13
  • Interleukin-2
  • Interleukin-6
  • Killer Cells, Natural
  • Luciferases
  • Lung Neoplasms
  • Lymphocyte Activation
  • Lymphocyte Depletion
  • Lymphocyte Transfusion
  • Lymphocytes
  • Lymphoma
  • Lymphopenia
  • Macrophages
  • Male
  • Membrane Glycoproteins
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Inbred CBA
  • Mice, Knockout
  • Mice, Mutant Strains
  • Mice, Nude
  • Mice, Transgenic
  • Microsomes
  • Middle Aged
  • Mitosis
  • Molecular Sequence Data
  • Myelodysplastic Syndromes
  • Myeloid Cells
  • Myeloid Differentiation Factor 88
  • NK Cell Lectin-Like Receptor Subfamily K
  • Neoplasms
  • North Carolina
  • Oocytes
  • Peripheral Blood Stem Cell Transplantation
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • RNA, Messenger
  • Receptors, Cell Surface
  • Receptors, Interleukin-1
  • Receptors, KIR
  • Recombinant Proteins
  • Retrospective Studies
  • Reverse Transcriptase Polymerase Chain Reaction
  • Risk Factors
  • STAT1 Transcription Factor
  • Sequence Deletion
  • Stem Cell Transplantation
  • Survival Rate
  • T-Cell Antigen Receptor Specificity
  • T-Lymphocytes
  • T-Lymphocytes, Cytotoxic
  • T-Lymphocytes, Regulatory
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Toll-Like Receptor 8
  • Toll-Like Receptor 9
  • Toll-Like Receptors
  • Transfection
  • Transgenes
  • Transplantation Conditioning
  • Transplantation, Homologous
  • Tumor Escape
  • Vaccines
  • Vaccinia
  • Vaccinia virus
  • Virus Diseases
  • Viruses
  • Xenopus
  • beta-Galactosidase