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Shinohara, Mari L.

Overview:

We need to mount a strong immune response against pathogens during infections, but excessive and uncontrolled immune reactions can lead to autoimmunity.  How does our immune system keep the balance fine-tuned?  This is a central question being asked in my laboratory.

Immune system needs to detect pathogens quickly and effectively.  This is performed by the innate immune system, which includes cells such as macrophages and dendritic cells (DCs).  Pathogens are recognized by pattern recognition receptors (PRRs) and may be cleared in the innate immune system.  However, when pathogens cannot be eliminated by innate immunity, the adaptive immune system participates by exploiting the ability of T cells and B cells.  The two immune systems work together not only to clear pathogens effectively but also to avoid collateral damages by from our own immune responses.

In my lab, we use mouse models for infectious and autoimmune diseases to understand the cellular and molecular mechanisms of; pathogen recognition by PRRs in macrophages and DCs, initiation of inflammatory responses in the innate immune system, and the impact of innate immune inflammation on the development and regulation of T cell-mediated adaptive immune responses.

Several projects are ongoing in the lab.  They are; (1) elucidating the role of the NLRP3 inflammasome, an innate immune sensor of pathogens and endogenous danger signals, in T-cell mediated pathology of EAE (an animal model of multiple sclerosis), (2) dissecting molecular mechanisms of pathogen recognition through Toll-like receptors (TLRs) and c-type lectin receptors (CLRs) and of downregulating  hyperinflammation, (3) molecular and cellular mechanisms in the innate immune system to induce immune tolerance in T cells, and (4) elucidating a role of a protein termed osteopontin (OPN), as both secreted (sOPN) and intracellular (iOPN) isoforms, in regulation of immune responses during infections and tumor development.  Although we are very active in EAE to study autoimmunity, other mouse models, such as psoriasis and colitis are ongoing.  As for infections, we are interested in fungal infections, which have not been well explored as bacterial and viral infections.  Cell types we study are mainly DCs, macrophages, and T cells.  By focusing on these immune cell types, we study impacts of infections on the development of autoimmunity.

Positions:

Associate Professor of Immunology

Immunology
School of Medicine

Assistant Professor in Molecular Genetics and Microbiology

Molecular Genetics and Microbiology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

News:

Publications:

Inflammasome activation in multiple sclerosis and experimental autoimmune encephalomyelitis (EAE).

The aptly named inflammasomes are powerful signaling complexes that sense inflammatory signals under a myriad of conditions, including those from infections and endogenous sources. The inflammasomes promote inflammation by maturation and release of the pro-inflammatory cytokines, IL-1β and IL-18. Several inflammasomes have been identified so far, but this review focuses mainly on the NLRP3 inflammasome. By still ill-defined activation mechanisms, a sensor molecule, NLRP3 (NACHT, LRR and PYD domains-containing protein 3), responds to danger signals and rapidly recruits ASC (apoptosis-associated speck-like protein containing a CARD) and pro-caspase-1 to form a large oligomeric signaling platform-the inflammasome. Involvement of the NLRP3 inflammasome in infections, metabolic disorders, autoinflammation, and autoimmunity, underscores its position as a central player in sensing microbial and damage signals and coordinating pro-inflammatory immune responses. Indeed, evidence in patients with multiple sclerosis (MS) suggests inflammasome activation occurs during disease. Experiments with the mouse model of MS, experimental autoimmune encephalomyelitis (EAE), specifically describe the NLRP3 inflammasome as critical and necessary to disease development. This review discusses recent studies in EAE and MS which describe associations of inflammasome activation with promotion of T cell pathogenicity, infiltration of cells into the central nervous system (CNS) and direct neurodegeneration during EAE and MS.

Authors
Barclay, W; Shinohara, ML
MLA Citation
Barclay, W, and Shinohara, ML. "Inflammasome activation in multiple sclerosis and experimental autoimmune encephalomyelitis (EAE)." Brain pathology (Zurich, Switzerland) 27.2 (March 2017): 213-219.
PMID
27997058
Source
epmc
Published In
Brain Pathology
Volume
27
Issue
2
Publish Date
2017
Start Page
213
End Page
219
DOI
10.1111/bpa.12477

Metabolic Alterations Contribute to Enhanced Inflammatory Cytokine Production in Irgm1-deficient Macrophages.

The immunity-related GTPases (IRGs) are a family of proteins that are induced by interferon (IFN)-γ and play pivotal roles in immune and inflammatory responses. IRGs ostensibly function as dynamin-like proteins that bind to intracellular membranes and promote remodeling and trafficking of those membranes. Prior studies have shown that loss of Irgm1 in mice leads to increased lethality to bacterial infections as well as enhanced inflammation to non-infectious stimuli; however, the mechanisms underlying these phenotypes are unclear. In the studies reported here, we found that uninfected Irgm1-deficient mice displayed high levels of serum cytokines typifying profound autoinflammation. Similar increases in cytokine production were also seen in cultured, IFN-γ-primed macrophages that lacked Irgm1. A series of metabolic studies indicated that the enhanced cytokine production was associated with marked metabolic changes in the Irgm1-deficient macrophages, including increased glycolysis and an accumulation of long chain acylcarnitines. Cells were exposed to the glycolytic inhibitor, 2-deoxyglucose, or fatty acid synthase inhibitors to perturb the metabolic alterations, which resulted in dampening of the excessive cytokine production. These results suggest that Irgm1 deficiency drives metabolic dysfunction in macrophages in a manner that is cell-autonomous and independent of infectious triggers. This may be a significant contributor to excessive inflammation seen in Irgm1-deficient mice in different contexts.

Authors
Schmidt, EA; Fee, BE; Henry, SC; Nichols, AG; Shinohara, ML; Rathmell, JC; MacIver, NJ; Coers, J; Ilkayeva, OR; Koves, TR; Taylor, GA
MLA Citation
Schmidt, EA, Fee, BE, Henry, SC, Nichols, AG, Shinohara, ML, Rathmell, JC, MacIver, NJ, Coers, J, Ilkayeva, OR, Koves, TR, and Taylor, GA. "Metabolic Alterations Contribute to Enhanced Inflammatory Cytokine Production in Irgm1-deficient Macrophages." The Journal of biological chemistry 292.11 (March 2017): 4651-4662.
PMID
28154172
Source
epmc
Published In
The Journal of biological chemistry
Volume
292
Issue
11
Publish Date
2017
Start Page
4651
End Page
4662
DOI
10.1074/jbc.m116.770735

An interferon-β-resistant and NLRP3 inflammasome-independent subtype of EAE with neuronal damage.

Inflammation induced by innate immunity influences the development of T cell-mediated autoimmunity in multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). We found that strong activation of innate immunity induced Nod-like receptor protein 3 (NLRP3) inflammasome-independent and interferon-β (IFNβ)-resistant EAE (termed type B EAE), whereas EAE induced by weak activation of innate immunity requires the NLRP3 inflammasome and is sensitive to IFNβ treatment. Instead, an alternative inflammatory mechanism, including membrane-bound lymphotoxin-β receptor (LTβR) and CXC chemokine receptor 2 (CXCR2), is involved in type B EAE development, and type B EAE is ameliorated by antagonizing these receptors. Relative expression of Ltbr and Cxcr2 genes was indeed enhanced in patients with IFNβ-resistant multiple sclerosis. Remission was minimal in type B EAE due to neuronal damages induced by semaphorin 6B upregulation on CD4+ T cells. Our data reveal a new inflammatory mechanism by which an IFNβ-resistant EAE subtype develops.

Authors
Inoue, M; Chen, P-H; Siecinski, S; Li, Q-J; Liu, C; Steinman, L; Gregory, SG; Benner, E; Shinohara, ML
MLA Citation
Inoue, M, Chen, P-H, Siecinski, S, Li, Q-J, Liu, C, Steinman, L, Gregory, SG, Benner, E, and Shinohara, ML. "An interferon-β-resistant and NLRP3 inflammasome-independent subtype of EAE with neuronal damage." Nature neuroscience 19.12 (December 2016): 1599-1609.
PMID
27820602
Source
epmc
Published In
Nature Neuroscience
Volume
19
Issue
12
Publish Date
2016
Start Page
1599
End Page
1609
DOI
10.1038/nn.4421

Osteopontin has a protective role in prostate tumor development in mice.

Osteopontin (OPN) is a protein, generally considered to play a pro-tumorigenic role, whereas several reports have demonstrated the anti-tumorigenic function of OPN during tumor development. These opposing anti- and pro-tumorigenic functions are not fully understood. Here, we report that host-derived OPN plays an anti-tumorigenic role in the transgenic adenocarcinoma of the mouse prostate (TRAMP) model and a TRAMP tumor transplant model. Tumor suppression mediated by OPN in Rag2-/- mice suggests that OPN is dispensable in the adaptive immune response. We found that host-derived OPN enhanced infiltration of natural killer (NK) cells into TRAMP tumors. The requirement of OPN in NK cell migration towards TRAMP cells was confirmed by an ex vivo cell migration assay. In contrast to TRAMP cells, in vivo B16 tumor development was not inhibited by OPN, and B16 tumors did not show OPN-mediated cell recruitment. It is possible that low levels of chemokine expression by B16 cells do not allow OPN to enhance immune cell recruitment. In addition to demonstrating the anti-tumorigenic role of OPN in TRAMP tumor development, this study also suggests that the contribution of OPN to tumor development depends on the type of tumor as well as the source and isoform of OPN.

Authors
Danzaki, K; Kanayama, M; Alcazar, O; Shinohara, ML
MLA Citation
Danzaki, K, Kanayama, M, Alcazar, O, and Shinohara, ML. "Osteopontin has a protective role in prostate tumor development in mice." European journal of immunology 46.11 (November 2016): 2669-2678.
PMID
27601131
Source
epmc
Published In
European Journal of Immunology
Volume
46
Issue
11
Publish Date
2016
Start Page
2669
End Page
2678
DOI
10.1002/eji.201646391

Lung inflammation stalls Th17-cell migration en route to the central nervous system during the development of experimental autoimmune encephalomyelitis.

Recruiting pathogenic T cells to the central nervous system (CNS) is a critical step during the development of experimental autoimmune encephalomyelitis (EAE). Here, we report that the absence of autophagy and microtubule-associated protein 1A/1B-light chain 3-associated phagocytosis significantly delayed the onset of EAE in Atg7 conditional knockout (Atg7 CKO) mice in myeloid cells. T-helper cell-cell priming appeared to be normal in the Atg7 CKO mice, but the mice showed significant accumulation of Th17 cells in the lung. The data suggested that the stalling of Th17 cells in the lung en route to the CNS caused the delay. The lung of Atg7 CKO mice, in which we previously demonstrated spontaneous mild inflammation, showed high expression of CCL20, a chemokine that attracts Th17 cells. We have also shown that LPS intranasal instillation delayed EAE onset, suggesting that pulmonary inflammation has an impact on EAE development. Based on our data, therapeutic immunomodulation targeted to the lung, rather than systemically, might be a possible future option to treat multiple sclerosis.

Authors
Kanayama, M; Danzaki, K; He, Y-W; Shinohara, ML
MLA Citation
Kanayama, M, Danzaki, K, He, Y-W, and Shinohara, ML. "Lung inflammation stalls Th17-cell migration en route to the central nervous system during the development of experimental autoimmune encephalomyelitis." International immunology 28.9 (September 2016): 463-469.
PMID
26989091
Source
epmc
Published In
International Immunology
Volume
28
Issue
9
Publish Date
2016
Start Page
463
End Page
469
DOI
10.1093/intimm/dxw013

Leptin directly promotes T-cell glycolytic metabolism to drive effector T-cell differentiation in a mouse model of autoimmunity.

Upon activation, T cells require energy for growth, proliferation, and function. Effector T (Teff) cells, such as Th1 and Th17 cells, utilize high levels of glycolytic metabolism to fuel proliferation and function. In contrast, Treg cells require oxidative metabolism to fuel suppressive function. It remains unknown how Teff/Treg-cell metabolism is altered when nutrients are limited and leptin levels are low. We therefore examined the role of malnutrition and associated hypoleptinemia on Teff versus Treg cells. We found that both malnutrition-associated hypoleptinemia and T cell-specific leptin receptor knockout suppressed Teff-cell number, function, and glucose metabolism, but did not alter Treg-cell metabolism or suppressive function. Using the autoimmune mouse model EAE, we confirmed that fasting-induced hypoleptinemia altered Teff-cell, but not Treg-cell, glucose metabolism, and function in vivo, leading to decreased disease severity. To explore potential mechanisms, we examined HIF-1α, a key regulator of Th17 differentiation and Teff-cell glucose metabolism, and found HIF-1α expression was decreased in T cell-specific leptin receptor knockout Th17 cells, and in Teff cells from fasted EAE mice, but was unchanged in Treg cells. Altogether, these data demonstrate a selective, cell-intrinsic requirement for leptin to upregulate glucose metabolism and maintain function in Teff, but not Treg cells.

Authors
Gerriets, VA; Danzaki, K; Kishton, RJ; Eisner, W; Nichols, AG; Saucillo, DC; Shinohara, ML; MacIver, NJ
MLA Citation
Gerriets, VA, Danzaki, K, Kishton, RJ, Eisner, W, Nichols, AG, Saucillo, DC, Shinohara, ML, and MacIver, NJ. "Leptin directly promotes T-cell glycolytic metabolism to drive effector T-cell differentiation in a mouse model of autoimmunity." European journal of immunology 46.8 (August 2016): 1970-1983.
Website
http://hdl.handle.net/10161/12472
PMID
27222115
Source
epmc
Published In
European Journal of Immunology
Volume
46
Issue
8
Publish Date
2016
Start Page
1970
End Page
1983
DOI
10.1002/eji.201545861

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).

Authors
Klionsky, DJ; Abdelmohsen, K; Abe, A; Abedin, MJ; Abeliovich, H; Acevedo Arozena, A; Adachi, H; Adams, CM; Adams, PD; Adeli, K; Adhihetty, PJ; Adler, SG; Agam, G; Agarwal, R; Aghi, MK; Agnello, M; Agostinis, P; Aguilar, PV; Aguirre-Ghiso, J; Airoldi, EM; Ait-Si-Ali, S; Akematsu, T; Akporiaye, ET; Al-Rubeai, M; Albaiceta, GM; Albanese, C; Albani, D; Albert, ML; Aldudo, J; Algül, H; Alirezaei, M; Alloza, I; Almasan, A; Almonte-Beceril, M; Alnemri, ES; Alonso, C; Altan-Bonnet, N; Altieri, DC et al.
MLA Citation
Klionsky, DJ, Abdelmohsen, K, Abe, A, Abedin, MJ, Abeliovich, H, Acevedo Arozena, A, Adachi, H, Adams, CM, Adams, PD, Adeli, K, Adhihetty, PJ, Adler, SG, Agam, G, Agarwal, R, Aghi, MK, Agnello, M, Agostinis, P, Aguilar, PV, Aguirre-Ghiso, J, Airoldi, EM, Ait-Si-Ali, S, Akematsu, T, Akporiaye, ET, Al-Rubeai, M, Albaiceta, GM, Albanese, C, Albani, D, Albert, ML, Aldudo, J, Algül, H, Alirezaei, M, Alloza, I, Almasan, A, Almonte-Beceril, M, Alnemri, ES, Alonso, C, Altan-Bonnet, N, and Altieri, DC et al. "Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)." Autophagy 12.1 (January 2016): 1-222.
PMID
26799652
Source
epmc
Published In
Autophagy
Volume
12
Issue
1
Publish Date
2016
Start Page
1
End Page
222
DOI
10.1080/15548627.2015.1100356

Roles of Autophagy and Autophagy-Related Proteins in Antifungal Immunity.

Autophagy was initially characterized as a process to digest cellular components, including damaged cell organelles or unused proteins. However, later studies showed that autophagy plays an important role to protect hosts from microbial infections. Accumulating evidences showed the contribution of autophagy itself and autophagy-related proteins (ATGs) in the clearance of bacteria, virus, and parasites. A number of studies also revealed the molecular mechanisms by which autophagy is initiated and developed. Furthermore, it is now understood that some ATGs are shared between two distinct processes; autophagy and LC3-associated phagocytosis (LAP). Thus, our understanding on autophagy has been greatly enhanced in the last decade. By contrast, roles of autophagy and ATGs in fungal infections are still elusive relative to those in bacterial and viral infections. Based on limited numbers of reports, ATG-mediated host responses appear to significantly vary depending on invading fungal species. In this review, we discuss how autophagy and ATGs are involved in antifungal immune responses based on recent discoveries.

Authors
Kanayama, M; Shinohara, ML
MLA Citation
Kanayama, M, and Shinohara, ML. "Roles of Autophagy and Autophagy-Related Proteins in Antifungal Immunity." Frontiers in immunology 7 (January 2016): 47-. (Review)
PMID
26925060
Source
epmc
Published In
Frontiers in Immunology
Volume
7
Publish Date
2016
Start Page
47
DOI
10.3389/fimmu.2016.00047

Calcineurin orchestrates dimorphic transitions, antifungal drug responses and host-pathogen interactions of the pathogenic mucoralean fungus Mucor circinelloides.

Calcineurin plays essential roles in virulence and growth of pathogenic fungi and is a target of the natural products FK506 and Cyclosporine A. In the pathogenic mucoralean fungus Mucor circinelloides, calcineurin mutation or inhibition confers a yeast-locked phenotype indicating that calcineurin governs the dimorphic transition. Genetic analysis in this study reveals that two calcineurin A catalytic subunits (out of three) are functionally diverged. Homology modeling illustrates modes of resistance resulting from amino substitutions in the interface between each calcineurin subunit and the inhibitory drugs. In addition, we show how the dimorphic transition orchestrated by calcineurin programs different outcomes during host-pathogen interactions. For example, when macrophages phagocytose Mucor yeast, subsequent phagosomal maturation occurs, indicating host cells respond appropriately to control the pathogen. On the other hand, upon phagocytosis of spores, macrophages fail to form mature phagosomes. Cytokine production from immune cells differs following exposure to yeast versus spores (which germinate into hyphae). Thus, the morphogenic transition can be targeted as an efficient treatment option against Mucor infection. In addition, genetic analysis (including gene disruption and mutational studies) further strengthens the understanding of calcineurin and provides a foundation to develop antifungal agents targeting calcineurin to deploy against Mucor and other pathogenic fungi.

Authors
Lee, SC; Li, A; Calo, S; Inoue, M; Tonthat, NK; Bain, JM; Louw, J; Shinohara, ML; Erwig, LP; Schumacher, MA; Ko, DC; Heitman, J
MLA Citation
Lee, SC, Li, A, Calo, S, Inoue, M, Tonthat, NK, Bain, JM, Louw, J, Shinohara, ML, Erwig, LP, Schumacher, MA, Ko, DC, and Heitman, J. "Calcineurin orchestrates dimorphic transitions, antifungal drug responses and host-pathogen interactions of the pathogenic mucoralean fungus Mucor circinelloides." Molecular microbiology 97.5 (September 2015): 844-865.
PMID
26010100
Source
epmc
Published In
Molecular Microbiology
Volume
97
Issue
5
Publish Date
2015
Start Page
844
End Page
865
DOI
10.1111/mmi.13071

Hyperinflammation, T cells, and endotoxemia.

Authors
Inoue, M; Shinohara, ML
MLA Citation
Inoue, M, and Shinohara, ML. "Hyperinflammation, T cells, and endotoxemia." Oncotarget 6.27 (September 2015): 23040-23041.
PMID
26309079
Source
epmc
Published In
Oncotarget
Volume
6
Issue
27
Publish Date
2015
Start Page
23040
End Page
23041

[NLRP3 inflammasome and multiple sclerosis/EAE].

Authors
Inoue, M; Shinohara, ML
MLA Citation
Inoue, M, and Shinohara, ML. "[NLRP3 inflammasome and multiple sclerosis/EAE]." Nihon rinsho. Japanese journal of clinical medicine 73 Suppl 7 (September 2015): 835-841.
PMID
26480802
Source
epmc
Published In
Nippon rinsho. Japanese journal of clinical medicine
Volume
73 Suppl 7
Publish Date
2015
Start Page
835
End Page
841

Cutting edge: Role of osteopontin and integrin αv in T cell-mediated anti-inflammatory responses in endotoxemia.

The immune system is equipped with mechanisms that downregulate hyperinflammation to avoid collateral damage. We demonstrated recently that unprimed T cells downregulate macrophage TNF production through direct interaction with macrophages in the spleen during LPS endotoxemia. How T cell migration toward macrophages occurs upon LPS injection is still not clear. In this study, we demonstrate that secreted osteopontin (sOPN) plays a role in the T cell migration to initiate the suppression of hyperinflammation during endotoxemia. Osteopontin levels in splenic macrophages were upregulated 2 h after LPS treatment, whereas T cell migration toward macrophages was observed 3 h after treatment. Neutralization of sOPN and blockade of its receptor, integrin αv, significantly inhibited CD4(+) T cell migration and increased susceptibility to endotoxemia. Our study demonstrates that the sOPN/integrin αv axis, which induces T cell chemotaxis toward macrophages, is critical for suppressing hyperinflammation during the first 3 h of endotoxemia.

Authors
Inoue, M; Shinohara, ML
MLA Citation
Inoue, M, and Shinohara, ML. "Cutting edge: Role of osteopontin and integrin αv in T cell-mediated anti-inflammatory responses in endotoxemia." Journal of immunology (Baltimore, Md. : 1950) 194.12 (June 2015): 5595-5598.
PMID
25972484
Source
epmc
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
194
Issue
12
Publish Date
2015
Start Page
5595
End Page
5598
DOI
10.4049/jimmunol.1500623

The lung is protected from spontaneous inflammation by autophagy in myeloid cells.

The lung is constantly exposed to the outer environment; thus, it must maintain a state of immune ignorance or tolerance not to overrespond to harmless environmental stimuli. How cells in the lung control immune responses under nonpathogenic condition is not fully understood. In this study, we found that autophagy plays a critical role in the lung-specific immune regulation that prevents spontaneous inflammation. Autophagy in pulmonary myeloid cells plays a role in maintaining low burdens of environmental microbes in the lung, as well as in lowering mitochondrial reactive oxygen species production and preventing overresponse to TLR4 ligands in alveolar macrophages. Based on these mechanisms, we also found that intranasal instillation of antibiotics or an inhibitor of reactive oxygen species was efficient in preventing spontaneous pulmonary inflammation. Thus, autophagy in myeloid cells, particularly alveolar macrophages, is critical for inhibiting spontaneous pulmonary inflammation, and pulmonary inflammation caused by dysfunctional autophagy is pharmacologically prevented.

Authors
Kanayama, M; He, Y-W; Shinohara, ML
MLA Citation
Kanayama, M, He, Y-W, and Shinohara, ML. "The lung is protected from spontaneous inflammation by autophagy in myeloid cells." Journal of immunology (Baltimore, Md. : 1950) 194.11 (June 2015): 5465-5471.
PMID
25911758
Source
epmc
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
194
Issue
11
Publish Date
2015
Start Page
5465
End Page
5471
DOI
10.4049/jimmunol.1403249

Neuroinflammation in Alzheimer's disease.

Increasing evidence suggests that Alzheimer's disease pathogenesis is not restricted to the neuronal compartment, but includes strong interactions with immunological mechanisms in the brain. Misfolded and aggregated proteins bind to pattern recognition receptors on microglia and astroglia, and trigger an innate immune response characterised by release of inflammatory mediators, which contribute to disease progression and severity. Genome-wide analysis suggests that several genes that increase the risk for sporadic Alzheimer's disease encode factors that regulate glial clearance of misfolded proteins and the inflammatory reaction. External factors, including systemic inflammation and obesity, are likely to interfere with immunological processes of the brain and further promote disease progression. Modulation of risk factors and targeting of these immune mechanisms could lead to future therapeutic or preventive strategies for Alzheimer's disease.

Authors
Heneka, MT; Carson, MJ; El Khoury, J; Landreth, GE; Brosseron, F; Feinstein, DL; Jacobs, AH; Wyss-Coray, T; Vitorica, J; Ransohoff, RM; Herrup, K; Frautschy, SA; Finsen, B; Brown, GC; Verkhratsky, A; Yamanaka, K; Koistinaho, J; Latz, E; Halle, A; Petzold, GC; Town, T; Morgan, D; Shinohara, ML; Perry, VH; Holmes, C; Bazan, NG; Brooks, DJ; Hunot, S; Joseph, B; Deigendesch, N; Garaschuk, O; Boddeke, E; Dinarello, CA; Breitner, JC; Cole, GM; Golenbock, DT; Kummer, MP
MLA Citation
Heneka, MT, Carson, MJ, El Khoury, J, Landreth, GE, Brosseron, F, Feinstein, DL, Jacobs, AH, Wyss-Coray, T, Vitorica, J, Ransohoff, RM, Herrup, K, Frautschy, SA, Finsen, B, Brown, GC, Verkhratsky, A, Yamanaka, K, Koistinaho, J, Latz, E, Halle, A, Petzold, GC, Town, T, Morgan, D, Shinohara, ML, Perry, VH, Holmes, C, Bazan, NG, Brooks, DJ, Hunot, S, Joseph, B, Deigendesch, N, Garaschuk, O, Boddeke, E, Dinarello, CA, Breitner, JC, Cole, GM, Golenbock, DT, and Kummer, MP. "Neuroinflammation in Alzheimer's disease." The Lancet. Neurology 14.4 (April 2015): 388-405.
PMID
25792098
Source
epmc
Published In
The Lancet Neurology
Volume
14
Issue
4
Publish Date
2015
Start Page
388
End Page
405
DOI
10.1016/s1474-4422(15)70016-5

Autophagy enhances NFκB activity in specific tissue macrophages by sequestering A20 to boost antifungal immunity.

Immune responses must be well restrained in a steady state to avoid excessive inflammation. However, such restraints are quickly removed to exert antimicrobial responses. Here we report a role of autophagy in an early host antifungal response by enhancing NFκB activity through A20 sequestration. Enhancement of NFκB activation is achieved by autophagic depletion of A20, an NFκB inhibitor, in F4/80(hi) macrophages in the spleen, peritoneum and kidney. We show that p62, an autophagic adaptor protein, captures A20 to sequester it in the autophagosome. This allows the macrophages to release chemokines to recruit neutrophils. Indeed, mice lacking autophagy in myeloid cells show higher susceptibility to Candida albicans infection due to impairment in neutrophil recruitment. Thus, at least in the specific aforementioned tissues, autophagy appears to break A20-dependent suppression in F4/80(hi) macrophages, which express abundant A20 and contribute to the initiation of efficient innate immune responses.

Authors
Kanayama, M; Inoue, M; Danzaki, K; Hammer, G; He, Y-W; Shinohara, ML
MLA Citation
Kanayama, M, Inoue, M, Danzaki, K, Hammer, G, He, Y-W, and Shinohara, ML. "Autophagy enhances NFκB activity in specific tissue macrophages by sequestering A20 to boost antifungal immunity." Nature communications 6 (January 22, 2015): 5779-.
Website
http://hdl.handle.net/10161/9376
PMID
25609235
Source
epmc
Published In
Nature Communications
Volume
6
Publish Date
2015
Start Page
5779
DOI
10.1038/ncomms6779

Metabolic programming and PDHK1 control CD4+ T cell subsets and inflammation.

Activation of CD4+ T cells results in rapid proliferation and differentiation into effector and regulatory subsets. CD4+ effector T cell (Teff) (Th1 and Th17) and Treg subsets are metabolically distinct, yet the specific metabolic differences that modify T cell populations are uncertain. Here, we evaluated CD4+ T cell populations in murine models and determined that inflammatory Teffs maintain high expression of glycolytic genes and rely on high glycolytic rates, while Tregs are oxidative and require mitochondrial electron transport to proliferate, differentiate, and survive. Metabolic profiling revealed that pyruvate dehydrogenase (PDH) is a key bifurcation point between T cell glycolytic and oxidative metabolism. PDH function is inhibited by PDH kinases (PDHKs). PDHK1 was expressed in Th17 cells, but not Th1 cells, and at low levels in Tregs, and inhibition or knockdown of PDHK1 selectively suppressed Th17 cells and increased Tregs. This alteration in the CD4+ T cell populations was mediated in part through ROS, as N-acetyl cysteine (NAC) treatment restored Th17 cell generation. Moreover, inhibition of PDHK1 modulated immunity and protected animals against experimental autoimmune encephalomyelitis, decreasing Th17 cells and increasing Tregs. Together, these data show that CD4+ subsets utilize and require distinct metabolic programs that can be targeted to control specific T cell populations in autoimmune and inflammatory diseases.

Authors
Gerriets, VA; Kishton, RJ; Nichols, AG; Macintyre, AN; Inoue, M; Ilkayeva, O; Winter, PS; Liu, X; Priyadharshini, B; Slawinska, ME; Haeberli, L; Huck, C; Turka, LA; Wood, KC; Hale, LP; Smith, PA; Schneider, MA; MacIver, NJ; Locasale, JW; Newgard, CB; Shinohara, ML; Rathmell, JC
MLA Citation
Gerriets, VA, Kishton, RJ, Nichols, AG, Macintyre, AN, Inoue, M, Ilkayeva, O, Winter, PS, Liu, X, Priyadharshini, B, Slawinska, ME, Haeberli, L, Huck, C, Turka, LA, Wood, KC, Hale, LP, Smith, PA, Schneider, MA, MacIver, NJ, Locasale, JW, Newgard, CB, Shinohara, ML, and Rathmell, JC. "Metabolic programming and PDHK1 control CD4+ T cell subsets and inflammation." The Journal of clinical investigation 125.1 (January 2015): 194-207.
Website
http://hdl.handle.net/10161/10313
PMID
25437876
Source
epmc
Published In
Journal of Clinical Investigation
Volume
125
Issue
1
Publish Date
2015
Start Page
194
End Page
207
DOI
10.1172/jci76012

T cells down-regulate macrophage TNF production by IRAK1-mediated IL-10 expression and control innate hyperinflammation.

Endotoxemia is caused by excessive inflammation, but the immune system has various mechanisms to avoid collateral organ damage in endotoxemia. A handful of reports have shown that innate immune responses are suppressed by the adaptive immune system. However, the molecular mechanism by which adaptive immune cells suppress innate inflammatory responses is not clear. Here, we report that T cells are shown to interact with macrophages at the early stage of enodotoxemia and to prolong survival of mice through controlling TNF and IL-10 levels by macrophage CD40 stimulation. The cross-talk between CD40 and toll-like receptor (TLR4) signaling first mediates IL-1 receptor-associated kinase 1 (IRAK1) nuclear translocation and its binding to the IL-10 gene promoter in macrophages, without interfering with the NFκB pathway. IL-10 is then detected by macrophages in an autocrine fashion to destabilize Tnfa mRNA. To induce IRAK1-mediated IL-10 expression, signals from both CD40 and TLR4 are essential. CD40 signaling induces IRAK1 sumoylation in the presence of TNF receptor-associated factor 2 (TRAF2) and intracellular isoform of osteopontin (iOPN) whereas TLR4 signaling provides IFN regulatory factor 5 (IRF5) as a chaperone for sumoylated IRAK1 nuclear translocation. Interaction of T cells with macrophages was observed in the spleen in vivo after endotoxemia induction with LPS injection. Our study demonstrates a mechanistic basis for the immunosuppressive role of macrophage CD40 in LPS endotoxemia.

Authors
Inoue, M; Arikawa, T; Chen, Y-H; Moriwaki, Y; Price, M; Brown, M; Perfect, JR; Shinohara, ML
MLA Citation
Inoue, M, Arikawa, T, Chen, Y-H, Moriwaki, Y, Price, M, Brown, M, Perfect, JR, and Shinohara, ML. "T cells down-regulate macrophage TNF production by IRAK1-mediated IL-10 expression and control innate hyperinflammation." Proceedings of the National Academy of Sciences of the United States of America 111.14 (April 2014): 5295-5300.
PMID
24706909
Source
epmc
Published In
Proceedings of the National Academy of Sciences of USA
Volume
111
Issue
14
Publish Date
2014
Start Page
5295
End Page
5300
DOI
10.1073/pnas.1321427111

Microtubule acetylation amplifies p38 kinase signalling and anti-inflammatory IL-10 production.

Reversible acetylation of α-tubulin is an evolutionarily conserved modification in microtubule networks. Despite its prevalence, the physiological function and regulation of microtubule acetylation remain poorly understood. Here we report that macrophages challenged by bacterial lipopolysaccharides (LPS) undergo extensive microtubule acetylation. Suppression of LPS-induced microtubule acetylation by inactivating the tubulin acetyltransferase, MEC17, profoundly inhibits the induction of anti-inflammatory interleukin-10 (IL-10), a phenotype effectively reversed by an acetylation-mimicking α-tubulin mutant. Conversely, elevating microtubule acetylation by inhibiting the tubulin deacetylase, HDAC6, or stabilizing microtubules via Taxol stimulates IL-10 hyper-induction. Supporting the anti-inflammatory function of microtubule acetylation, HDAC6 inhibition significantly protects mice from LPS toxicity. In HDAC6-deficient macrophages challenged by LPS, p38 kinase signalling becomes selectively amplified, leading to SP1-dependent IL-10 transcription. Remarkably, the augmented p38 signalling is suppressed by MEC17 inactivation. Our findings identify reversible microtubule acetylation as a kinase signalling modulator and a key component in the inflammatory response.

Authors
Wang, B; Rao, Y-H; Inoue, M; Hao, R; Lai, C-H; Chen, D; McDonald, SL; Choi, M-C; Wang, Q; Shinohara, ML; Yao, T-P
MLA Citation
Wang, B, Rao, Y-H, Inoue, M, Hao, R, Lai, C-H, Chen, D, McDonald, SL, Choi, M-C, Wang, Q, Shinohara, ML, and Yao, T-P. "Microtubule acetylation amplifies p38 kinase signalling and anti-inflammatory IL-10 production." Nature communications 5 (March 17, 2014): 3479-.
PMID
24632940
Source
epmc
Published In
Nature Communications
Volume
5
Publish Date
2014
Start Page
3479
DOI
10.1038/ncomms4479

Clustering of pattern recognition receptors for fungal detection.

Authors
Inoue, M; Shinohara, ML
MLA Citation
Inoue, M, and Shinohara, ML. "Clustering of pattern recognition receptors for fungal detection." PLoS pathogens 10.2 (February 20, 2014): e1003873-.
PMID
24586145
Source
epmc
Published In
PLoS pathogens
Volume
10
Issue
2
Publish Date
2014
Start Page
e1003873
DOI
10.1371/journal.ppat.1003873

The role of interferon-β in the treatment of multiple sclerosis and experimental autoimmune encephalomyelitis - in the perspective of inflammasomes.

Inflammasomes in innate immune cells mediate the induction of inflammation by sensing microbes and pathogen-associated/damage-associated molecular patterns. Inflammasomes are also known to be involved in the development of some human and animal autoimmune diseases. The Nod-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is currently the most fully characterized inflammasome, although a limited number of studies have demonstrated its role in demyelinating autoimmune diseases in the central nervous system of humans and animals. Currently, the development of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS), is known to be induced by the NLRP3 inflammasome through enhanced recruitment of inflammatory immune cells in the central nervous system. On the other hand, interferon-β (IFNβ), a first-line drug to treat MS, inhibits NLRP3 inflammasome activation, and ameliorates EAE. The NLRP3 inflammasome is indeed a factor capable of inducing EAE, but it is dispensable when EAE is induced by aggressive disease induction regimens. In such NLRP3 inflammasome-independent EAE, IFN-β treatment is generally not effective. This might therefore be one mechanism that leads to occasional failures of IFN-β treatment in EAE, and possibly, in MS as well. In the current review, we discuss inflammasomes and autoimmunity; in particular, the impact of the NLRP3 inflammasome on MS/EAE, and on IFN-β therapy.

Authors
Inoue, M; Shinohara, ML
MLA Citation
Inoue, M, and Shinohara, ML. "The role of interferon-β in the treatment of multiple sclerosis and experimental autoimmune encephalomyelitis - in the perspective of inflammasomes." Immunology 139.1 (May 2013): 11-18. (Review)
PMID
23360426
Source
pubmed
Published In
Immunology
Volume
139
Issue
1
Publish Date
2013
Start Page
11
End Page
18
DOI
10.1111/imm.12081

Mnk1 and 2 are dispensable for T cell development and activation but important for the pathogenesis of experimental autoimmune encephalomyelitis.

T cell development and activation are usually accompanied by expansion and production of numerous proteins that require active translation. The eukaryotic translation initiation factor 4E (eIF4E) binds to the 5' cap structure of mRNA and is critical for cap-dependent translational initiation. It has been hypothesized that MAPK-interacting kinase 1 and 2 (Mnk1/2) promote cap-dependent translation by phosphorylating eIF4E at serine 209 (S209). Pharmacologic studies using inhibitors have suggested that Mnk1/2 have important roles in T cells. However, genetic evidence supporting such conclusions is lacking. Moreover, the signaling pathways that regulate Mnk1/2 in T cells remain unclear. We demonstrate that TCR engagement activates Mnk1/2 in primary T cells. Such activation is dependent on Ras-Erk1/2 signaling and is inhibited by diacylglycerol kinases α and ζ. Mnk1/2 double deficiency in mice abolishes TCR-induced eIF4E S209 phosphorylation, indicating their absolute requirement for eIF4E S209 phosphorylation. However, Mnk1/2 double deficiency does not affect the development of conventional αβ T cells, regulatory T cells, or NKT cells. Furthermore, T cell activation, in vivo primary and memory CD8 T cell responses to microbial infection, and NKT cell cytokine production were not obviously altered by Mnk1/2 deficiency. Although Mnk1/2 deficiency causes decreased IL-17 and IFN-γ production by CD4 T cells following immunization of mice with myelin oligodendrocyte glycoprotein peptide in complete Freund's adjuvant, correlating with milder experimental autoimmune encephalomyelitis scores, it does not affect Th cell differentiation in vitro. Together, these data suggest that Mnk1/2 has a minimal role in T cell development and activation but may regulate non-T cell lineages to control Th1 and Th17 differentiation in vivo.

Authors
Gorentla, BK; Krishna, S; Shin, J; Inoue, M; Shinohara, ML; Grayson, JM; Fukunaga, R; Zhong, X-P
MLA Citation
Gorentla, BK, Krishna, S, Shin, J, Inoue, M, Shinohara, ML, Grayson, JM, Fukunaga, R, and Zhong, X-P. "Mnk1 and 2 are dispensable for T cell development and activation but important for the pathogenesis of experimental autoimmune encephalomyelitis." J Immunol 190.3 (February 1, 2013): 1026-1037.
PMID
23269249
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
190
Issue
3
Publish Date
2013
Start Page
1026
End Page
1037
DOI
10.4049/jimmunol.1200026

NLRP3 Inflammasome and MS/EAE.

Inflammasomes are cytosolic sensors that detect pathogens and danger signals in the innate immune system. The NLRP3 inflammasome is currently the most fully characterized inflammasome and is known to detect a wide array of microbes and endogenous damage-associated molecules. Possible involvement of the NLRP3 inflammasome (or inflammasomes) in the development of multiple sclerosis (MS) was suggested in a number of studies. Recent studies showed that the NLRP3 inflammasome exacerbates experimental autoimmune encephalomyelitis (EAE), an animal model of MS, although EAE can also develop without the NLRP3 inflammasome. In this paper, we discuss the NLRP3 inflammasome in MS and EAE development.

Authors
Inoue, M; Shinohara, ML
MLA Citation
Inoue, M, and Shinohara, ML. "NLRP3 Inflammasome and MS/EAE." Autoimmune Dis 2013 (2013): 859145-.
PMID
23365725
Source
pubmed
Published In
Autoimmune Diseases
Volume
2013
Publish Date
2013
Start Page
859145
DOI
10.1155/2013/859145

Transcriptional regulator Id2 is required for the CD4 T cell immune response in the development of experimental autoimmune encephalomyelitis.

An effective immune response to Ag challenge is critically dependent on the size of the effector cell population generated from clonal activation of Ag-specific T cells. The transcription network involved in regulating the size of the effector population, particularly for CD4 Th cells, is poorly understood. In this study, we investigate the role of Id2, an inhibitor of E protein transcription factors, in the generation of CD4 effectors. Using a T cell-specific conditional Id2 knockout mouse model, we show that inhibitor of DNA binding (Id)2 is essential for the development of experimental autoimmune encephalomyelitis. Although Ag-specific and IL-17-producing CD4 T cells are produced in these mice, the activated CD4 T cells form a smaller pool of effector cells in the peripheral lymphoid organs, exhibit reduced proliferation and increased cell death, and are largely absent in the CNS. In the absence of Id2, E protein targets, including the proapoptotic protein Bim and SOCS3, are expressed at higher levels among activated CD4 T cells. This study reveals a critical role of Id2 in the control of effector CD4 T cell population size and the development of a Th17-mediated autoimmune disease.

Authors
Lin, Y-Y; Jones-Mason, ME; Inoue, M; Lasorella, A; Iavarone, A; Li, Q-J; Shinohara, ML; Zhuang, Y
MLA Citation
Lin, Y-Y, Jones-Mason, ME, Inoue, M, Lasorella, A, Iavarone, A, Li, Q-J, Shinohara, ML, and Zhuang, Y. "Transcriptional regulator Id2 is required for the CD4 T cell immune response in the development of experimental autoimmune encephalomyelitis." J Immunol 189.3 (August 1, 2012): 1400-1405.
PMID
22745378
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
189
Issue
3
Publish Date
2012
Start Page
1400
End Page
1405
DOI
10.4049/jimmunol.1200491

NLRP3 inflammasome induces chemotactic immune cell migration to the CNS in experimental autoimmune encephalomyelitis.

The NLRP3 inflammasome is a multiprotein complex consisting of three kinds of proteins, NLRP3, ASC, and pro-caspase-1, and plays a role in sensing pathogens and danger signals in the innate immune system. The NLRP3 inflammasome is thought to be involved in the development of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). However, the mechanism by which the NLRP3 inflammasome induces EAE is not clear. In this study, we found that the NLRP3 inflammasome played a critical role in inducing T-helper cell migration into the CNS. To gain migratory ability, CD4(+) T cells need to be primed by NLRP3 inflammasome-sufficient antigen-presenting cells to up-regulate chemotaxis-related proteins, such as osteopontin, CCR2, and CXCR6. In the presence of the NLRP3 inflammasome, dendritic cells and macrophages also induce chemotactic ability and up-regulate chemotaxis-related proteins, such as α4β1 integrin, CCL7, CCL8, and CXCL16. On the other hand, reduced Th17 cell population size in immunized Nlrp3(-/-) and Asc(-/-) mice is not a determinative factor for their resistance to EAE. As currently applied in clinical interventions of MS, targeting immune cell migration molecules may be an effective approach in treating MS accompanied by NLRP3 inflammasome activation.

Authors
Inoue, M; Williams, KL; Gunn, MD; Shinohara, ML
MLA Citation
Inoue, M, Williams, KL, Gunn, MD, and Shinohara, ML. "NLRP3 inflammasome induces chemotactic immune cell migration to the CNS in experimental autoimmune encephalomyelitis." Proc Natl Acad Sci U S A 109.26 (June 26, 2012): 10480-10485.
PMID
22699511
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
109
Issue
26
Publish Date
2012
Start Page
10480
End Page
10485
DOI
10.1073/pnas.1201836109

Interferon-β therapy against EAE is effective only when development of the disease depends on the NLRP3 inflammasome.

Interferon-β (IFN-β) is widely used to treat multiple sclerosis (MS), and its efficacy was demonstrated in the setting of experimental autoimmune encephalomyelitis (EAE), an animal model of MS; however, IFN-β is not effective in treating all cases of MS. Here, we demonstrate that signaling by IFNAR (the shared receptor for IFN-α and IFN-β) on macrophages inhibits activation of Rac1 and the generation of reactive oxygen species (ROS) through suppressor of cytokine signaling 1 (SOCS1). The inhibition of Rac1 activation and ROS generation suppressed the activity of the Nod-like receptor (NLR) family, pyrin domain-containing 3 (NLRP3) inflammasome, which resulted in attenuated EAE pathogenicity. We further found that two subsets of EAE could be defined on the basis of their dependency on the NLRP3 inflammasome and that IFN-β was not an effective therapy when EAE was induced in an NLRP3 inflammasome-independent fashion. Thus, our study demonstrates a previously uncharacterized signaling pathway that is involved in the suppression of EAE by IFN-β and characterizes NLRP3-independent EAE, which cannot be treated with IFN-β.

Authors
Inoue, M; Williams, KL; Oliver, T; Vandenabeele, P; Rajan, JV; Miao, EA; Shinohara, ML
MLA Citation
Inoue, M, Williams, KL, Oliver, T, Vandenabeele, P, Rajan, JV, Miao, EA, and Shinohara, ML. "Interferon-β therapy against EAE is effective only when development of the disease depends on the NLRP3 inflammasome. (Published online)" Sci Signal 5.225 (May 22, 2012): ra38-.
PMID
22623753
Source
pubmed
Published In
Science Signaling
Volume
5
Issue
225
Publish Date
2012
Start Page
ra38
DOI
10.1126/scisignal.2002767

M-HIFU inhibits tumor growth, suppresses STAT3 activity and enhances tumor specific immunity in a transplant tumor model of prostate cancer.

OBJECTIVE: In this study, we explored the use of mechanical high intensity focused ultrasound (M-HIFU) as a neo-adjuvant therapy prior to surgical resection of the primary tumor. We also investigated the role of signal transducer and activator of transcription 3 (STAT3) in M-HIFU elicited anti-tumor immune response using a transplant tumor model of prostate cancer. METHODS: RM-9, a mouse prostate cancer cell line with constitutively activated STAT3, was inoculated subcutaneously in C57BL/6J mice. The tumor-bearing mice (with a maximum tumor diameter of 5∼6 mm) were treated by M-HIFU or sham exposure two days before surgical resection of the primary tumor. Following recovery, if no tumor recurrence was observed in 30 days, tumor rechallenge was performed. The growth of the rechallenged tumor, survival rate and anti-tumor immune response of the animal were evaluated. RESULTS: No tumor recurrence and distant metastasis were observed in both treatment groups employing M-HIFU + surgery and surgery alone. However, compared to surgery alone, M-HIFU combined with surgery were found to significantly inhibit the growth of rechallenged tumors, down-regulate intra-tumoral STAT3 activities, increase cytotoxic T cells in spleens and tumor draining lymph nodes (TDLNs), and improve the host survival. Furthermore, M-HIFU combined with surgery was found to significantly decrease the level of immunosuppression with concomitantly increased number and activities of dendritic cells, compared to surgery alone. CONCLUSION: Our results demonstrate that M-HIFU can inhibit STAT3 activities, and when combined synergistically with surgery, may provide a novel and promising strategy for the treatment of prostate cancers.

Authors
Huang, X; Yuan, F; Liang, M; Lo, H-W; Shinohara, ML; Robertson, C; Zhong, P
MLA Citation
Huang, X, Yuan, F, Liang, M, Lo, H-W, Shinohara, ML, Robertson, C, and Zhong, P. "M-HIFU inhibits tumor growth, suppresses STAT3 activity and enhances tumor specific immunity in a transplant tumor model of prostate cancer." PLoS One 7.7 (2012): e41632-.
PMID
22911830
Source
pubmed
Published In
PloS one
Volume
7
Issue
7
Publish Date
2012
Start Page
e41632
DOI
10.1371/journal.pone.0041632

Estrogen-related receptor-α is a metabolic regulator of effector T-cell activation and differentiation.

Stimulation of resting CD4(+) T lymphocytes leads to rapid proliferation and differentiation into effector (Teff) or inducible regulatory (Treg) subsets with specific functions to promote or suppress immunity. Importantly, Teff and Treg use distinct metabolic programs to support subset specification, survival, and function. Here, we describe that the orphan nuclear receptor estrogen-related receptor-α (ERRα) regulates metabolic pathways critical for Teff. Resting CD4(+) T cells expressed low levels of ERRα protein that increased on activation. ERRα deficiency reduced activated T-cell numbers in vivo and cytokine production in vitro but did not seem to modulate immunity through inhibition of activating signals or viability. Rather, ERRα broadly affected metabolic gene expression and glucose metabolism essential for Teff. In particular, up-regulation of Glut1 protein, glucose uptake, and mitochondrial processes were suppressed in activated ERRα(-/-) T cells and T cells treated with two chemically independent ERRα inhibitors or by shRNAi. Acute ERRα inhibition also blocked T-cell growth and proliferation. This defect appeared as a result of inadequate glucose metabolism, because provision of lipids, but not increased glucose uptake or pyruvate, rescued ATP levels and cell division. Additionally, we have shown that Treg requires lipid oxidation, whereas Teff uses glucose metabolism, and lipid addition selectively restored Treg--but not Teff--generation after acute ERRα inhibition. Furthermore, in vivo inhibition of ERRα reduced T-cell proliferation and Teff generation in both immunization and experimental autoimmune encephalomyelitis models. Thus, ERRα is a selective transcriptional regulator of Teff metabolism that may provide a metabolic means to modulate immunity.

Authors
Michalek, RD; Gerriets, VA; Nichols, AG; Inoue, M; Kazmin, D; Chang, C-Y; Dwyer, MA; Nelson, ER; Pollizzi, KN; Ilkayeva, O; Giguere, V; Zuercher, WJ; Powell, JD; Shinohara, ML; McDonnell, DP; Rathmell, JC
MLA Citation
Michalek, RD, Gerriets, VA, Nichols, AG, Inoue, M, Kazmin, D, Chang, C-Y, Dwyer, MA, Nelson, ER, Pollizzi, KN, Ilkayeva, O, Giguere, V, Zuercher, WJ, Powell, JD, Shinohara, ML, McDonnell, DP, and Rathmell, JC. "Estrogen-related receptor-α is a metabolic regulator of effector T-cell activation and differentiation." Proc Natl Acad Sci U S A 108.45 (November 8, 2011): 18348-18353.
PMID
22042850
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
108
Issue
45
Publish Date
2011
Start Page
18348
End Page
18353
DOI
10.1073/pnas.1108856108

Intracellular osteopontin (iOPN) and immunity.

Osteopontin (OPN) is a protein involved in various pathophysiological events. OPN has been studied as a secreted protein, but recent reports showed that OPN can be found in the cytoplasm and the nucleus. Therefore, some OPN molecules are not secreted and stay in cells. Such intracellular OPN (iOPN) has biological functions distinct from secreted OPN (sOPN). iOPN is involved in cytoskeletal rearrangement and in signal transduction pathways downstream of innate immune receptors, such as Toll-like receptors (TLRs), as an adaptor or scaffolding protein. Although sOPN and iOPN are generated from the same Opn mRNA species, biological outcomes mediated by two isoforms can be different. It would be necessary to delineate which isoform of OPN is responsible for pathophysiological events.

Authors
Inoue, M; Shinohara, ML
MLA Citation
Inoue, M, and Shinohara, ML. "Intracellular osteopontin (iOPN) and immunity." Immunol Res 49.1-3 (April 2011): 160-172. (Review)
PMID
21136203
Source
pubmed
Published In
Immunologic Research
Volume
49
Issue
1-3
Publish Date
2011
Start Page
160
End Page
172
DOI
10.1007/s12026-010-8179-5

Osteopontin expression during early cerebral ischemia-reperfusion in rats: enhanced expression in the right cortex is suppressed by acetaminophen.

Osteopontin (OPN) is a pleiotropic protein implicated in various inflammatory responses including ischemia-reperfusion (I-R) injury. Two distinct forms of the protein have been identified: an extensively studied secreted form (sOPN) and a less-well-known intracellular form (iOPN). Studies have shown that increased OPN expression parallels the time course of macrophage infiltration into injured tissue, a late event in the development of cerebral infarcts. sOPN has been suggested to promote remodeling of the extracellular matrix in the brain; the function of iOPN may be to facilitate certain signal transduction processes. Here, we studied OPN expression in adult male Sprague-Dawley rats subjected to global forebrain I-R injury. We found iOPN in the cytoplasm of both cortices and the hippocampus, but unexpectedly only the right cortex exhibited a marked increase in the iOPN level after 45 min of reperfusion. Acetaminophen, a drug recently shown to decrease apoptotic incidence, caspase-9 activation, and mitochondrial dysfunction during global I-R, significantly inhibited the increase in iOPN protein in the right cortex, suggesting a role for iOPN in the response to I-R injury in the right cortex.

Authors
Baliga, SS; Merrill, GF; Shinohara, ML; Denhardt, DT
MLA Citation
Baliga, SS, Merrill, GF, Shinohara, ML, and Denhardt, DT. "Osteopontin expression during early cerebral ischemia-reperfusion in rats: enhanced expression in the right cortex is suppressed by acetaminophen. (Published online)" PLoS One 6.1 (January 21, 2011): e14568-.
PMID
21283687
Source
pubmed
Published In
PloS one
Volume
6
Issue
1
Publish Date
2011
Start Page
e14568
DOI
10.1371/journal.pone.0014568

Cutting edge: critical role of intracellular osteopontin in antifungal innate immune responses.

We found that absence of osteopontin (OPN) in immunocompromised Rag2(-/-) mice, which lack T and B cells, made the mice extremely susceptible to an opportunistic fungus Pneumocystis, although immunocompetent OPN-deficient mice could clear Pneumocystis as well as wild-type mice. OPN has been studied as an extracellular protein, and the role of an intracellular isoform of OPN (iOPN) is still largely unknown. In this study, we elucidated the mechanism by which iOPN was involved in antifungal innate immunity. First, iOPN was essential for cluster formation of fungal receptors that detect Pneumocystis, including dectin-1, TLR2, and mannose receptor. Second, iOPN played a role as an adaptor molecule in TLR2 and dectin-1 signaling pathways and mediated ERK activation and cytokine production by zymosan, which simultaneously activates TLR2 and dectin-1 pathways. Third, iOPN enhanced phagocytosis and clearance of Pneumocystis. Our study suggests the critical involvement of iOPN in antifungal innate immunity.

Authors
Inoue, M; Moriwaki, Y; Arikawa, T; Chen, Y-H; Oh, YJ; Oliver, T; Shinohara, ML
MLA Citation
Inoue, M, Moriwaki, Y, Arikawa, T, Chen, Y-H, Oh, YJ, Oliver, T, and Shinohara, ML. "Cutting edge: critical role of intracellular osteopontin in antifungal innate immune responses." J Immunol 186.1 (January 1, 2011): 19-23.
PMID
21135164
Source
pubmed
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
186
Issue
1
Publish Date
2011
Start Page
19
End Page
23
DOI
10.4049/jimmunol.1002735

Unexpected role of clathrin adaptor AP-1 in MHC-dependent positive selection of T cells.

Trafficking of transmembrane receptors to a specific intracellular compartment is conducted by adaptor molecules that bind to target motifs within the cytoplasmic domains of cargo proteins. We generated mice containing a lymphoid-specific deficiency of AP-1 using RNAi knockdown technology. Inhibition of AP-1 expression in thymocytes blocks progression from double-positive immature thymocytes, resulting in complete absence of CD4(+) single-positive thymocytes and severe reduction of CD3(+)CD8(+) single-positive thymocytes. Analysis of the contribution of AP-1 deficiency on the interaction between mature CD4(+) T cells and antigen-presenting cells revealed that AP-1 is essential to efficient immune synapse formation and associated T cell activation, suggesting a possible mechanism of AP-1 function in thymocyte development.

Authors
Alvarez Arias, DA; McCarty, N; Lu, L; Maldonado, RA; Shinohara, ML; Cantor, H
MLA Citation
Alvarez Arias, DA, McCarty, N, Lu, L, Maldonado, RA, Shinohara, ML, and Cantor, H. "Unexpected role of clathrin adaptor AP-1 in MHC-dependent positive selection of T cells." Proc Natl Acad Sci U S A 107.6 (February 9, 2010): 2556-2561.
PMID
20133794
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
107
Issue
6
Publish Date
2010
Start Page
2556
End Page
2561
DOI
10.1073/pnas.0913671107

Reprogramming after chromosome transfer into mouse blastomeres.

It is well known that oocytes can reprogram differentiated cells, allowing animal cloning by nuclear transfer. We have recently shown that fertilized zygotes retain reprogramming activities, suggesting that such activities might also persist in cleavage-stage embryos. Here, we used chromosome transplantation techniques to investigate whether the blastomeres of two-cell-stage mouse embryos can reprogram more differentiated cells. When chromosomes from one of the two blastomeres were replaced with the chromosomes of an embryonic or CD4(+) T lymphocyte donor cell, we observed nuclear reprogramming and efficient contribution of the manipulated cell to the developing blastocyst. Embryos produced by this method could be used to derive stem cell lines and also developed to term, generating mosaic "cloned" animals. These results demonstrate that blastomeres retain reprogramming activities and support the notion that discarded human preimplantation embryos may be useful recipients for the production of genetically tailored human embryonic stem cell lines.

Authors
Egli, D; Sandler, VM; Shinohara, ML; Cantor, H; Eggan, K
MLA Citation
Egli, D, Sandler, VM, Shinohara, ML, Cantor, H, and Eggan, K. "Reprogramming after chromosome transfer into mouse blastomeres." Curr Biol 19.16 (August 25, 2009): 1403-1409.
PMID
19682906
Source
pubmed
Published In
Current Biology
Volume
19
Issue
16
Publish Date
2009
Start Page
1403
End Page
1409
DOI
10.1016/j.cub.2009.06.065

Engagement of the type I interferon receptor on dendritic cells inhibits T helper 17 cell development: role of intracellular osteopontin.

Mechanisms that prevent inappropriate or excessive interleukin-17-producing T helper (Th17) cell responses after microbial infection may be necessary to avoid autoimmunity. Here, we define a pathway initiated by engagement of type I IFN receptor (IFNAR) expressed by dendritic cells (DC) that culminated in suppression of Th17 cell differentiation. IFNAR-dependent inhibition of an intracellular translational isoform of Osteopontin, termed Opn-i, derepressed interleukin-27 (IL-27) secretion and prevented efficient Th17 responses. Moreover, Opn-i expression in DC and microglia regulated the type and intensity of experimental autoimmune encephalomyelitis (EAE). Mice containing DC deficient in Opn-i produced excessive amounts of IL-27 and developed a delayed disease characterized by an enhanced Th1 response compared with the dominant Th17 response of Opn-sufficient mice. Definition of the IFNAR-Opn-i axis that controls Th17 development provides insight into regulation of Th cell sublineage development and the molecular basis of type I interferon therapy for MS and other autoimmune diseases.

Authors
Shinohara, ML; Kim, J-H; Garcia, VA; Cantor, H
MLA Citation
Shinohara, ML, Kim, J-H, Garcia, VA, and Cantor, H. "Engagement of the type I interferon receptor on dendritic cells inhibits T helper 17 cell development: role of intracellular osteopontin." Immunity 29.1 (July 18, 2008): 68-78.
PMID
18619869
Source
pubmed
Published In
Immunity
Volume
29
Issue
1
Publish Date
2008
Start Page
68
End Page
78
DOI
10.1016/j.immuni.2008.05.008

Alternative translation of osteopontin generates intracellular and secreted isoforms that mediate distinct biological activities in dendritic cells.

Osteopontin (Opn) contributes to diverse biological processes that include immune responses, vascularization, and bone formation. Until recently, studies describing the activities of Opn have focused on the cytokine-like properties of the secreted protein. Here, we show that alternative translation of a single Opn mRNA species generates a secreted and intracellular isoform. Utilization of a 5' canonical translation start site generates a protein that includes an N-terminal signal sequence allowing targeting to secretory vesicles and cytokine secretion, whereas usage of a downstream start site generates a shortened protein that lacks the N-terminal signal sequence and localizes mainly to cytoplasm. The coordinated action of these Opn gene products regulates the functional phenotype of subsets of dendritic cells.

Authors
Shinohara, ML; Kim, H-J; Kim, J-H; Garcia, VA; Cantor, H
MLA Citation
Shinohara, ML, Kim, H-J, Kim, J-H, Garcia, VA, and Cantor, H. "Alternative translation of osteopontin generates intracellular and secreted isoforms that mediate distinct biological activities in dendritic cells." Proc Natl Acad Sci U S A 105.20 (May 20, 2008): 7235-7239.
PMID
18480255
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
105
Issue
20
Publish Date
2008
Start Page
7235
End Page
7239
DOI
10.1073/pnas.0802301105

The bridge between dendritic cells and asthma.

Authors
Shinohara, ML; Cantor, H
MLA Citation
Shinohara, ML, and Cantor, H. "The bridge between dendritic cells and asthma." Nat Med 13.5 (May 2007): 536-538.
PMID
17479095
Source
pubmed
Published In
Nature Medicine
Volume
13
Issue
5
Publish Date
2007
Start Page
536
End Page
538
DOI
10.1038/nm0507-536

Innate immune mechanisms that promote development of effector and regulatory CD4 lineages in EAE (Experimental Autoimmune Encephalomyelitis)

Authors
Shinohara, ML; Cantor, H
MLA Citation
Shinohara, ML, and Cantor, H. "Innate immune mechanisms that promote development of effector and regulatory CD4 lineages in EAE (Experimental Autoimmune Encephalomyelitis)." September 2006.
Source
wos-lite
Published In
Journal of Neuroimmunology
Volume
178
Publish Date
2006
Start Page
34
End Page
34

Osteopontin expression is essential for interferon-alpha production by plasmacytoid dendritic cells.

The observation that the T-bet transcription factor allows tissue-specific upregulation of intracellular osteopontin (Opn-i) in plasmacytoid dendritic cells (pDCs) suggests that Opn might contribute to the expression of interferon-alpha (IFN-alpha) in those cells. Here we show that Opn deficiency substantially reduced Toll-like receptor 9 (TLR9)-dependent IFN-alpha responses but spared expression of transcription factor NF-kappaB-dependent proinflammatory cytokines. Shortly after TLR9 engagement, colocalization of Opn-i and the adaptor molecule MyD88 was associated with induction of transcription factor IRF7-dependent IFN-alpha gene expression, whereas deficient expression of Opn-i was associated with defective nuclear translocation of IRF7 in pDCs. The importance of the Opn-IFN-alpha pathway was emphasized by its essential involvement in cross-presentation in vitro and in anti-herpes simplex virus 1 IFN-alpha response in vivo. The finding that Opn-i selectively coupled TLR9 signaling to expression of IFN-alpha but not to that of other proinflammatory cytokines provides new molecular insight into the biology of pDCs.

Authors
Shinohara, ML; Lu, L; Bu, J; Werneck, MBF; Kobayashi, KS; Glimcher, LH; Cantor, H
MLA Citation
Shinohara, ML, Lu, L, Bu, J, Werneck, MBF, Kobayashi, KS, Glimcher, LH, and Cantor, H. "Osteopontin expression is essential for interferon-alpha production by plasmacytoid dendritic cells." Nat Immunol 7.5 (May 2006): 498-506.
PMID
16604075
Source
pubmed
Published In
Nature Immunology
Volume
7
Issue
5
Publish Date
2006
Start Page
498
End Page
506
DOI
10.1038/ni1327

Osteopontin expression is essential for interferon-α production by plasmacytoid dendritic cells

Authors
SHINOHARA, ML
MLA Citation
SHINOHARA, ML. "Osteopontin expression is essential for interferon-α production by plasmacytoid dendritic cells." Nat. Immunol. 7 (2006): 498-506.
Source
cinii-english
Published In
Nat. Immunol.
Volume
7
Publish Date
2006
Start Page
498
End Page
506

Osteopontin expression is essential for interferon-α production by plasmacytoid dendritic cells

Authors
SHINOHARA, M
MLA Citation
SHINOHARA, M. "Osteopontin expression is essential for interferon-α production by plasmacytoid dendritic cells." Nat. Immunol. 7 (2006): 498-506.
Source
cinii-english
Published In
Nat. Immunol.
Volume
7
Publish Date
2006
Start Page
498
End Page
506
DOI
10.1038/ni1327

T-bet-dependent expression of osteopontin contributes to T cell polarization.

The osteopontin (Opn) glycoprotein has been implicated in diverse physiological processes, including vascularization, bone formation, and inflammatory responses. Studies of its role in immune responses has suggested that Opn can set the early stage of type-1 immune (cell-mediated) responses through differential regulation of IL-12 and IL-10 cytokine gene expression in macrophages. Although Opn has been suggested to play a role in the development of type-1 immunity, little is known about control of Opn gene expression. Here, we report that Opn gene expression in activated T cells, but not macrophages, is regulated by T-bet, a transcription factor that controls CD4+ T helper (Th1) cell lineage commitment. We also find that T-bet-dependent expression of Opn in T cells is essential for efficient skewing of CD4+ T and CD8+ T cells toward the Th1 and type 1 CD8+ T cells (Tc1) pathway, respectively. Taken together, these findings begin to delineate the genetic basis of Opn expression in T cells and further clarify the role of Opn in Th and Tc1 development.

Authors
Shinohara, ML; Jansson, M; Hwang, ES; Werneck, MBF; Glimcher, LH; Cantor, H
MLA Citation
Shinohara, ML, Jansson, M, Hwang, ES, Werneck, MBF, Glimcher, LH, and Cantor, H. "T-bet-dependent expression of osteopontin contributes to T cell polarization." Proc Natl Acad Sci U S A 102.47 (November 22, 2005): 17101-17106.
PMID
16286640
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
102
Issue
47
Publish Date
2005
Start Page
17101
End Page
17106
DOI
10.1073/pnas.0508666102

Lack of requirement of osteopontin for inflammation, bone erosion, and cartilage damage in the K/BxN model of autoantibody-mediated arthritis.

OBJECTIVE: Osteopontin (OPN) is a secreted glycoprotein involved in a range of physiologic processes, including inflammation, immunity mediated by Th1 cells, and bone remodeling. It is expressed in the joints of rheumatoid arthritis patients and has been the subject of conflicting reports concerning its role in arthritis induced by antibodies against type II collagen. This study assessed the role of OPN in the K/BxN serum-transfer model of autoantibody-induced arthritis. METHODS: Expression of OPN gene transcripts was assessed by microarray analysis of ankle RNA taken at 6 time points after transfer of K/BxN serum. OPN-sufficient or OPN-deficient littermates backcrossed for 10 generations onto the C57BL/6 genetic background were given K/BxN serum. Arthritis severity was measured by ankle thickening and a clinical index. Hind limb sections were stained with hematoxylin and eosin or toluidine blue and scored for inflammation, cartilage damage, and bone erosion. RESULTS: OPN messenger RNA transcripts progressively increased in ankle joints during the course of K/BxN serum-transferred arthritis. OPN-deficient mice receiving K/BxN serum developed arthritis with kinetics and clinical severity comparable with those of OPN-sufficient littermates. Histologic assessment of arthritic joints from OPN-deficient mice revealed synovial hyperplasia, pannus formation, mononuclear cell infiltration, bone erosion, cartilage damage at sites adjacent to and distal from pannus invasion, and tartrate-resistant acid phosphatase-positive multinucleated cells at sites of bone erosion. Histopathologic scoring demonstrated comparable levels of inflammation, cartilage damage, and bone erosion in OPN-sufficient and OPN-deficient mice. CONCLUSION: OPN does not have a required role in inflammation, bone erosion, and cartilage damage in the K/BxN serum-transfer model.

Authors
Jacobs, JP; Pettit, AR; Shinohara, ML; Jansson, M; Cantor, H; Gravallese, EM; Mathis, D; Benoist, C
MLA Citation
Jacobs, JP, Pettit, AR, Shinohara, ML, Jansson, M, Cantor, H, Gravallese, EM, Mathis, D, and Benoist, C. "Lack of requirement of osteopontin for inflammation, bone erosion, and cartilage damage in the K/BxN model of autoantibody-mediated arthritis." Arthritis Rheum 50.8 (August 2004): 2685-2694.
PMID
15334485
Source
pubmed
Published In
Arthritis and Rheumatism
Volume
50
Issue
8
Publish Date
2004
Start Page
2685
End Page
2694
DOI
10.1002/art.20381

Detailed analysis of gene expression during development of T cell lineages in the thymus.

The genetic mechanisms that promote lineage commitment and eliminate autoreactive cells in the thymus are not well understood. To better understand this process, we have identified and quantitated transcripts in the two major thymocyte lineages by using serial analysis of gene expression. Approximately 25 genes displayed almost complete segregation to one or the other T cell lineage. Commitment to the CD4 lineage was marked by up-regulation of genes associated with increased survival and chaperone function followed by expression of genes that regulate nucleosome remodeling and T cell receptor signaling. Differentiation within the CD8 lineage, on the other hand, was marked by up-regulation of genes that regulate lymphocyte homing, followed by quenching of genes that inhibit apoptosis. Definition of differential gene expression during development of the two major thymocyte lineages will allow insight into mechanisms of T cell development after positive and negative selection.

Authors
McCarty, N; Shinohara, ML; Lu, L; Cantor, H
MLA Citation
McCarty, N, Shinohara, ML, Lu, L, and Cantor, H. "Detailed analysis of gene expression during development of T cell lineages in the thymus." Proc Natl Acad Sci U S A 101.25 (June 22, 2004): 9339-9344.
PMID
15190182
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
101
Issue
25
Publish Date
2004
Start Page
9339
End Page
9344
DOI
10.1073/pnas.0402654101

Analysis of regulatory CD8 T cells in Qa-1-deficient mice.

The mouse protein Qa-1 (HLA-E in humans) is essential for immunological protection and immune regulation. Although Qa-1 has been linked to CD8 T cell-dependent suppression, the physiological relevance of this observation is unclear. We generated mice deficient in Qa-1 to develop an understanding of this process. Qa-1-deficient mice develop exaggerated secondary CD4 responses to foreign and self peptides. Enhanced responses to proteolipid protein self peptide were associated with resistance of Qa-1-deficient CD4 T cells to Qa-1-restricted CD8 T suppressor activity and increased susceptibility to experimental autoimmune encephalomyelitis. These findings delineate a Qa-1-dependent T cell-T cell inhibitory interaction that prevents the pathogenic expansion of autoreactive CD4 T cell populations and consequent autoimmune disease.

Authors
Hu, D; Ikizawa, K; Lu, L; Sanchirico, ME; Shinohara, ML; Cantor, H
MLA Citation
Hu, D, Ikizawa, K, Lu, L, Sanchirico, ME, Shinohara, ML, and Cantor, H. "Analysis of regulatory CD8 T cells in Qa-1-deficient mice." Nat Immunol 5.5 (May 2004): 516-523.
PMID
15098030
Source
pubmed
Published In
Nature Immunology
Volume
5
Issue
5
Publish Date
2004
Start Page
516
End Page
523
DOI
10.1038/ni1063

Neurospora clock-controlled gene 9 (ccg-9) encodes trehalose synthase: circadian regulation of stress responses and development.

The circadian clock of Neurospora crassa regulates the rhythmic expression of a number of genes encoding diverse functions which, as an ensemble, are adaptive to life in a rhythmic environment of alternating levels of light and dark, warmth and coolness, and dryness and humidity. Previous differential screens have identified a number of such genes based solely on their cycling expression, including clock-controlled gene 9 (ccg-9). Sequence analysis now shows the predicted CCG-9 polypeptide to be homologous to a novel form of trehalose synthase; as such it would catalyze the synthesis of the disaccharide trehalose, which plays an important role in protecting many cells from environmental stresses. Consistent with this, heat, glucose starvation, and osmotic stress induce ccg-9 transcript accumulation. Surprisingly, however, a parallel role in development is suggested by the finding that inactivation of ccg-9 results in altered conidiophore morphology and abolishes the normal circadian rhythm of asexual macroconidial development. Examination of a clock component, FRQ, in the ccg-9-null strain revealed normal cycling, phosphorylation, and light induction, indicating that loss of the conidiation rhythm is not due to changes in either the circadian oscillator or light input into the clock but pointing instead to a defect in circadian output. These data imply an interplay between a role of trehalose in stress protection and an apparent requirement for trehalose in clock regulation of conidiation under constant environmental conditions. This requirement can be bypassed by a daily light signal which drives a light-entrained rhythm in conidiation in the ccg-9-null strain; this bypass suggests that the trehalose requirement is related to clock control of development and not to the developmental process itself. Circadian control of trehalose synthase suggests a link between clock control of stress responses and that of development.

Authors
Shinohara, ML; Correa, A; Bell-Pedersen, D; Dunlap, JC; Loros, JJ
MLA Citation
Shinohara, ML, Correa, A, Bell-Pedersen, D, Dunlap, JC, and Loros, JJ. "Neurospora clock-controlled gene 9 (ccg-9) encodes trehalose synthase: circadian regulation of stress responses and development." Eukaryot Cell 1.1 (February 2002): 33-43.
PMID
12455969
Source
pubmed
Published In
Eukaryotic cell
Volume
1
Issue
1
Publish Date
2002
Start Page
33
End Page
43

A novel thermostable branching enzyme from an extremely thermophilic bacterial species, Rhodothermus obamensis.

A branching enzyme (EC 2.4.1.18) gene was isolated from an extremely thermophilic bacterium, Rhodothermus obamensis. The predicted protein encodes a polypeptide of 621 amino acids with a predicted molecular mass of 72 kDa. The deduced amino acid sequence shares 42-50% similarity to known bacterial branching enzyme sequences. Similar to the Bacillus branching enzymes, the predicted protein has a shorter N-terminal amino acid extension than that of the Escherichia coli branching enzyme. The deduced amino acid sequence does not appear to contain a signal sequence, suggesting that it is an intracellular enzyme. The R. obamensis branching enzyme was successfully expressed both in E. coli and a filamentous fungus, Aspergillus oryzae. The enzyme showed optimum catalytic activity at pH 6.0-6.5 and 65 degrees C. The enzyme was stable after 30 min at 80 degrees C and retained 50% of activity at 80 degrees C after 16 h. Branching activity of the enzyme was higher toward amylose than toward amylopectin. This is the first thermostable branching enzyme isolated from an extreme thermophile.

Authors
Shinohara, ML; Ihara, M; Abo, M; Hashida, M; Takagi, S; Beck, TC
MLA Citation
Shinohara, ML, Ihara, M, Abo, M, Hashida, M, Takagi, S, and Beck, TC. "A novel thermostable branching enzyme from an extremely thermophilic bacterial species, Rhodothermus obamensis." Appl Microbiol Biotechnol 57.5-6 (December 2001): 653-659.
PMID
11778874
Source
pubmed
Published In
Applied Microbiology and Biotechnology
Volume
57
Issue
5-6
Publish Date
2001
Start Page
653
End Page
659

LMP2 expression and proteasome activity in NOD mice.

Authors
Kessler, BM; Lennon-Duménil, AM; Shinohara, ML; Lipes, MA; Ploegh, HL
MLA Citation
Kessler, BM, Lennon-Duménil, AM, Shinohara, ML, Lipes, MA, and Ploegh, HL. "LMP2 expression and proteasome activity in NOD mice." Nat Med 6.10 (October 2000): 1064-. (Letter)
PMID
11017111
Source
pubmed
Published In
Nature Medicine
Volume
6
Issue
10
Publish Date
2000
Start Page
1064
DOI
10.1038/80346

LMP2 expression and proteasome activity in NOD mice [1] (multiple letters)

Authors
Kessler, BM; Lennon-Duménil, A-M; Shinohara, ML; Lipes, MA; Ploegh, HL
MLA Citation
Kessler, BM, Lennon-Duménil, A-M, Shinohara, ML, Lipes, MA, and Ploegh, HL. "LMP2 expression and proteasome activity in NOD mice [1] (multiple letters)." Nature Medicine 6.10 (2000): 1064-1066.
Source
scival
Published In
Nature Medicine
Volume
6
Issue
10
Publish Date
2000
Start Page
1064
End Page
1066

Comparison of ribosomal DNA ITS regions among geographic isolates of Cenococcum geophilum.

Cenococcum geophilum is an ecologically important mycorrhizal fungus with a global distribution and a wide host range. It has been difficult to study since it forms only sterile mycelia and, occasionally, sclerotial bodies. Because of its lack of morphological variability, its taxonomy and phylogenetic origins have until recently remained unclear. To better understand the genetic variation and environmental adaptability of C. geophilum, a molecular phylogeny was constructed based on the nuclear ribosomal DNA internal transcribed spacers (ITS1 and ITS2) of 69 isolates from various hosts and habitats. The results suggest DNA sequence conservation in the ITS regions. Considering its broad geographic and host range, this ITS conservation was unexpected. Our data imply that the ITS2 region is under evolutionary pressure to maintain the RNA secondary structure (similar to the pressure on the CgSSU introns) involved in the post-transcriptional processing of rRNA. Also, C. geophilum has very short ITS regions, thus limiting the number of mutable sites. This limited ITS variability suggests a recent radiation of C. geophilum, having been geographically distributed by a variety of efficient processes. C. geophilum appears to be a single taxonomic entity, possibly a single species. Therefore, it is an extremely adaptable, as well as ecologically valuable, taxon.

Authors
Shinohara, ML; LoBuglio, KF; Rogers, SO
MLA Citation
Shinohara, ML, LoBuglio, KF, and Rogers, SO. "Comparison of ribosomal DNA ITS regions among geographic isolates of Cenococcum geophilum." Curr Genet 35.5 (June 1999): 527-535.
PMID
10369960
Source
pubmed
Published In
Current Genetics
Volume
35
Issue
5
Publish Date
1999
Start Page
527
End Page
535

Glyceraldehyde-3-phosphate dehydrogenase is regulated on a daily basis by the circadian clock.

Circadian clocks function to govern a wide range of rhythmic activities in organisms. An integral part of rhythmicity is the daily control of target genes by the clock. Here we describe the sequence and analysis of a novel clock-controlled gene, ccg-7, showing similarity to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme widely used as a constitutive control in a variety of systems. That ccg-7 encodes GAPDH was confirmed by demonstrating that in vitro synthesized CCG-7 possesses GAPDH activity. Rhythms in both ccg-7 mRNA accumulation and CCG-7 (GAPDH) activity are observed in a clock wild-type strain where the peak in GAPDH activity lags several hours behind the peak in ccg-7 mRNA accumulation in the late night. Together with our previous observation that ccg-7 mRNA is not developmentally regulated, we show that ccg-7 is not induced by environmental stresses such as glucose or nitrogen deprivation (which also trigger development), heat shock, or osmotic stress. Thus, the finding that GAPDH is clock-regulated points to a specific role for the circadian clock in controlling aspects of general metabolism and provides evidence for circadian regulation of a gene found in most living organisms.

Authors
Shinohara, ML; Loros, JJ; Dunlap, JC
MLA Citation
Shinohara, ML, Loros, JJ, and Dunlap, JC. "Glyceraldehyde-3-phosphate dehydrogenase is regulated on a daily basis by the circadian clock." J Biol Chem 273.1 (January 2, 1998): 446-452.
PMID
9417102
Source
pubmed
Published In
The Journal of biological chemistry
Volume
273
Issue
1
Publish Date
1998
Start Page
446
End Page
452

Circadian clock-controlled genes isolated from Neurospora crassa are late night- to early morning-specific.

An endogenous circadian biological clock controls the temporal aspects of life in most organisms, including rhythmic control of genes involved in clock output pathways. In the fungus Neurospora crassa, one pathway known to be under control of the clock is asexual spore (conidia) development. To understand more fully the processes that are regulated by the N. crassa circadian clock, systematic screens were carried out for genes that oscillate at the transcriptional level. Time-of-day-specific cDNA libraries were generated and used in differential screens to identify six new clock-controlled genes (ccgs). Transcripts specific for each of the ccgs preferentially accumulate during the late night to early morning, although they vary with respect to steady-state mRNA levels and amplitude of the rhythm. Sequencing of the ends of the new ccg cDNAs revealed that ccg-12 is identical to N. crassa cmt encoding copper metallothionein, providing the suggestion that not all clock-regulated genes in N. crassa are specifically involved in the development of conidia. This was supported by finding that half of the new ccgs, including cmt(ccg-12), are not transcriptionally induced by developmental or light signals. These data suggest a major role for the clock in the regulation of biological processes distinct from development.

Authors
Bell-Pedersen, D; Shinohara, ML; Loros, JJ; Dunlap, JC
MLA Citation
Bell-Pedersen, D, Shinohara, ML, Loros, JJ, and Dunlap, JC. "Circadian clock-controlled genes isolated from Neurospora crassa are late night- to early morning-specific." Proc Natl Acad Sci U S A 93.23 (November 12, 1996): 13096-13101.
PMID
8917550
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
93
Issue
23
Publish Date
1996
Start Page
13096
End Page
13101

Group-I intron family in the nuclear ribosomal RNA small subunit genes of Cenococcum geophilum isolates.

A family of optional group-I introns was found near the 3' end of the nuclear small subunit rRNA genes in 61 out of 70 isolates of the deuteromycete mycorrhizal fungus Cenococcum geophilum. DNA sequence polymorphisms among the introns (termed CgSSU introns) from ten of the isolates were studied. The sequences, ranging in size from 488 to 514 nucleotides, were from 93.2% to 99.6% similar to each other. Mutations were less common in predicted base-paired regions (33% of all mutations) than in free-standing regions (67%). The introns were self-spliced in vitro and were closest to subgroup IC1 according to sequence and predicted secondary structure. Group-I intron pairing regions P1 through P10, including core regions P, Q, R and S, were present in all ten CgSSU introns studied. No lengthy open reading frames were found in any of the introns, indicating that the introns do not encode a protein, and therefore may not be mobile. It is likely that a single intron entered a progenote of C. geophilum and changed as the species evolved.

Authors
Shinohara, ML; LoBuglio, KF; Rogers, SO
MLA Citation
Shinohara, ML, LoBuglio, KF, and Rogers, SO. "Group-I intron family in the nuclear ribosomal RNA small subunit genes of Cenococcum geophilum isolates." Curr Genet 29.4 (March 1996): 377-387.
PMID
8598059
Source
pubmed
Published In
Current Genetics
Volume
29
Issue
4
Publish Date
1996
Start Page
377
End Page
387

The genetic and molecular dissection of a prototypic circadian system

A great deal is known about this archetypal circadian system, and it is likely that Neurospora will represent the first circadian system in which it will be possible to provide a complete description of the flow of information from the photoreceptor, through the components of oscillator, out to a terminal aspect of regulation. In Neurospora the strongest case has been made for there being a state variable of clock identified (Hall, 1995), it has now been shown that light resetting of the clock is mediated by the rapid light induction of the gene encoding this state variable, and a number of defined clock-regulated output genes have been identified, in two of which the clock-specific parts of the promoters have been localized. In addition to the importance of these factoids themselves, our efforts towards understanding of this system has allowed the development of tools and paradigms (e.g. Loros et al., 1989; Loros and Dunlap, 1991; Aronson et al., 1994a) that will help to pave the way for proving the identity of clock components in more complex systems, for understanding how clocks are regulated by entraining factors, and for showing how time information eventually is used to regulate the behaviors of clock cells, and of whole organisms.

Authors
Dunlap, JC; Loros, JJ; Merrow, M; Crosthwaite, S; Bell-Pedersen, D; Garceau, N; Shinohara, M; Cho, H; Luo, C
MLA Citation
Dunlap, JC, Loros, JJ, Merrow, M, Crosthwaite, S, Bell-Pedersen, D, Garceau, N, Shinohara, M, Cho, H, and Luo, C. "The genetic and molecular dissection of a prototypic circadian system." Progress in Brain Research 111 (1996): 11-27.
PMID
8990904
Source
scival
Published In
Progress in Brain Research
Volume
111
Publish Date
1996
Start Page
11
End Page
27

Messenger RNA intron in the nuclear 18 s ribosomal RNA gene of deuteromycetes

Introns within messenger RNA genes have characteristic border sequences and a conserved region near the 3′ end of the intron. All are involved in splicing to produce the mature mRNA. Introns in ribosomal RNA genes have less well-defined borders and contain no internal conservation. We report here mRNA-type introns located near the 3′ end of the 18 s rRNA genes of the deuteromycetes Phialophora americana and Cenococcum geophilum. Inserted sequences of various sizes have also been located at the same point in several other deuteromycete species. © 1993 Springer-Verlag.

Authors
Rogers, SO; Yan, ZH; Shinohara, M; LoBuglio, KF; Wang, CJK
MLA Citation
Rogers, SO, Yan, ZH, Shinohara, M, LoBuglio, KF, and Wang, CJK. "Messenger RNA intron in the nuclear 18 s ribosomal RNA gene of deuteromycetes." Current Genetics 23.4 (1993): 338-342.
PMID
8467532
Source
scival
Published In
Current Genetics
Volume
23
Issue
4
Publish Date
1993
Start Page
338
End Page
342
DOI
10.1007/BF00310896

Regulation of T-helper-cell lineage development by osteopontin: the inside story.

Studies of osteopontin (OPN)-dependent regulation of immune responses have focused on the cytokine activities of the secreted form of this protein. Recent evidence has revealed that an intracellular form of OPN expressed by dendritic cells regulates the expression of pro-inflammatory cytokines and the differentiation of T helper (T(H))-cell lineages. In this Opinion article, we discuss the properties of both OPN isoforms and their respective contributions to the immune response. We propose that cell-type-specific expression of secreted and intracellular OPN regulates the development of distinct effector T(H) cells, including that of T(H)1 and T(H)17 cells.

Authors
Cantor, H; Shinohara, ML
MLA Citation
Cantor, H, and Shinohara, ML. "Regulation of T-helper-cell lineage development by osteopontin: the inside story." Nat Rev Immunol 9.2: 137-141. (Review)
PMID
19096390
Source
pubmed
Published In
Nature Reviews Immunology
Volume
9
Issue
2
Start Page
137
End Page
141
DOI
10.1038/nri2460
Show More

Research Areas:

  • 1,4-alpha-Glucan Branching Enzyme
  • Acetaminophen
  • Adaptive Immunity
  • Adaptor Protein Complex 1
  • Adoptive Transfer
  • Alternative Splicing
  • Amino Acid Sequence
  • Aniline Compounds
  • Animals
  • Antigen-Presenting Cells
  • Arthritis
  • Aspergillus oryzae
  • Asthma
  • Astrocytes
  • Autoantibodies
  • Autoimmune Diseases
  • Autoimmunity
  • Biological Markers
  • Blotting, Western
  • Bone and Bones
  • Brain Injuries
  • Brain Ischemia
  • CD4-Positive T-Lymphocytes
  • CD8-Positive T-Lymphocytes
  • Caspase 1
  • Cell Adhesion
  • Cell Differentiation
  • Cell Line, Tumor
  • Cell Lineage
  • Cell Membrane
  • Cell Proliferation
  • Cell Separation
  • Cell Survival
  • Cells, Cultured
  • Central Nervous System
  • Cerebral Cortex
  • Chemotaxis
  • Chimera
  • Chromosomes
  • Circadian Rhythm
  • Clinical Trials as Topic
  • Cloning, Molecular
  • CpG Islands
  • Cross-Priming
  • Cysteine Endopeptidases
  • Cytochalasin B
  • Cytoplasm
  • Cytotoxicity, Immunologic
  • DNA Primers
  • DNA, Bacterial
  • DNA, Complementary
  • DNA, Ribosomal
  • Dendritic Cells
  • Diabetes Mellitus, Type 1
  • Disease Models, Animal
  • Embryo Transfer
  • Encephalomyelitis, Autoimmune, Experimental
  • Energy Metabolism
  • Enzyme Activation
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli
  • Eukaryotic Initiation Factor-4E
  • Evolution, Molecular
  • Female
  • Flow Cytometry
  • Fluorescent Dyes
  • Fungal Proteins
  • Fungi
  • G0 Phase
  • Gene Amplification
  • Gene Deletion
  • Gene Expression Regulation
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Enzymologic
  • Gene Expression Regulation, Fungal
  • Gene Knock-In Techniques
  • Gene Library
  • Genes, Bacterial
  • Genes, Fungal
  • Genetic Linkage
  • Genetic Predisposition to Disease
  • Genetic Variation
  • Genetics, Population
  • Genome, Fungal
  • Glucose
  • Glucosyltransferases
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • HLA Antigens
  • Herpesvirus 1, Human
  • High-Intensity Focused Ultrasound Ablation
  • Histocompatibility Antigens
  • Histocompatibility Antigens Class I
  • Histocompatibility Antigens Class II
  • Histones
  • Homeostasis
  • Humans
  • Immunity
  • Immunity, Innate
  • Immunization
  • Immunoblotting
  • Immunological Synapses
  • Inflammasomes
  • Inflammation
  • Inflammation Mediators
  • Inhibitor of Differentiation Protein 2
  • Integrins
  • Interferon-alpha
  • Interferon-beta
  • Interferon-gamma
  • Interleukin-17
  • Interleukin-1beta
  • Intracellular Fluid
  • Intracellular Space
  • Introns
  • Killer Cells, Natural
  • Lectins, C-Type
  • Lipopolysaccharides
  • Listeriosis
  • Luminescent Proteins
  • Lung
  • Lymph Nodes
  • Lymphocyte Activation
  • Lymphocyte Count
  • Lymphopoiesis
  • MAP Kinase Signaling System
  • Macrophages
  • Male
  • Membrane Proteins
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Inbred NOD
  • Mice, Knockout
  • Mice, Mutant Strains
  • Mice, Transgenic
  • Microglia
  • Mitochondria
  • Models, Biological
  • Models, Genetic
  • Models, Immunological
  • Molecular Sequence Data
  • Mosaicism
  • Multiple Sclerosis
  • Mycoses
  • Myelin-Oligodendrocyte Glycoprotein
  • Myeloid Cells
  • Natural Killer T-Cells
  • Nerve Tissue Proteins
  • Neuropeptides
  • Neurospora crassa
  • Nocodazole
  • Nucleic Acid Conformation
  • Oligonucleotide Array Sequence Analysis
  • Osteopontin
  • Ovalbumin
  • Oxidative Stress
  • Phagocytosis
  • Phenotype
  • Phosphorylation
  • Phylogeny
  • Plasmids
  • Pneumocystis
  • Pneumocystis Infections
  • Pneumonia
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Prostatic Neoplasms
  • Proteasome Endopeptidase Complex
  • Protein Isoforms
  • Protein Processing, Post-Translational
  • Protein Structure, Tertiary
  • Protein-Serine-Threonine Kinases
  • Proteins
  • Proto-Oncogene Proteins c-vav
  • RNA Caps
  • RNA Interference
  • RNA Splicing
  • RNA, Fungal
  • RNA, Messenger
  • RNA, Ribosomal
  • RNA, Small Nuclear
  • Rats
  • Reactive Oxygen Species
  • Receptor, Interferon alpha-beta
  • Receptors, Antigen, T-Cell
  • Receptors, Estrogen
  • Receptors, Pattern Recognition
  • Recombinant Proteins
  • Research Embryo Creation
  • Restriction Mapping
  • Reverse Transcriptase Polymerase Chain Reaction
  • STAT3 Transcription Factor
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Sialoglycoproteins
  • Signal Transduction
  • Spleen
  • Spores, Fungal
  • Suppressor of Cytokine Signaling Proteins
  • Survival Analysis
  • T-Box Domain Proteins
  • T-Lymphocyte Subsets
  • T-Lymphocytes
  • T-Lymphocytes, Cytotoxic
  • T-Lymphocytes, Helper-Inducer
  • T-Lymphocytes, Regulatory
  • Th1 Cells
  • Th17 Cells
  • Toll-Like Receptor 2
  • Toll-Like Receptors
  • Transcription Factors
  • Transcription, Genetic
  • Transcriptome
  • Virus Diseases
  • rac GTP-Binding Proteins
  • rac1 GTP-Binding Protein