Overview

The Duke Immune Profiling Core is a Duke School of Medicine Service Center and Duke Cancer Institute (DCI) Shared Resource whose academic home resides in the Department of Surgery.

Mission

Our overarching mission, to identify immunologic signatures that predict clinical outcomes, comprises two components.

Service

The first is a service component in which DIPC provides comprehensive, highly standardized, state-of-the-art human immunologic profiling services in support of both intra- and extramural translational research endeavors, including Phase I/II clinical trials.

Discovery

The second component focuses on discovery and utilizes advanced, high-dimensional Core technologies and resources to stimulate novel translational research initiatives that will impact existing standard medical care.

With over 35 years of experience in flow cytometry and immune profiling, we are available to support a wide range of services, such as small-scale projects for generating pilot data extending through comprehensive clinical trial testing. Additionally, our services extend to chip-based single-cell proteomics and multiplex immunoassays, study planning, performing experiments, data collection, analysis, and presentation support.

Services

  • Tissue processing and biorepository
  • Immune phenotyping by high-parameter polychromatic flow cytometry
  • Multiplex immunoassays
  • Chip-based single-cell proteomics assays (Isolight)
  • Bioinformatics and biostatistics
  • Computer stations for data analysis

DPIC Core Rates (PDF, 62.03 KB)

Equipment

BD SORP Symphony A5

The LSRFortessa X50 high dimensional analyzer has a five-laser bench (405nm 200mW, 488nm 400mW, 532nm 1000mW, 628nm 200mW, and 355nm 60mW) and is equipped with 44 fluorescence detectors, in addition to forward and side scatter detectors. Five decagons, coupled with the five higher-power lasers, tunable to variable power settings, allow for up to 28 parameters to be measured simultaneously. Laser Control Software allows for the adjustment of the individual laser powers as well as monitoring key performance parameters of the Coherent lasers. VXP electronics are configured for the detection of 46 parameters. The LSRFortessa X50 is equipped with a FACSFlow Supply System to regulate sample pressure accurately during high-volume use. The LSRFortessa X50 includes High Parameter Technical Applications Support as well as panel design assistance and support. The LSRFortessa X50 is maintained via an annual service contract.

BD SORP LSRFortessa (18 colors)

The SORP LSRFortessa analyzer has a five-laser bench (405nm 100mW, 488nm 50mW, 532nm 150mW, 640nm 100mW, and 355nm 20mW) and is equipped with 18 fluorescence detectors, in addition to forward and side scatter detectors. Two octagons and three trigons, coupled with the five lasers, allow for a total of 20 parameters to be measured simultaneously. Additional electronics have been added to the LSRFortessa to support the optical configuration. The LSRFortessa is equipped with a FACSFlow Supply System to regulate sample pressure accurately during high-volume use. The LSRFortessa is controlled by FACSDiva software that continuously samples signals at 10Mhz and processes signals to derive peak, width, and area measurements for each channel. The system has no electronic dead time and enables thresholding from any laser and multiple threshold parameters simultaneously. FACSDiva provides enhanced multicolor analysis and full compensation networks that allow the facility to acquire FCS files using digital technology. The LSRFortessa has an optical configuration to match the custom-designed LSRFortessa X50s for redundancy and backup as needed. The LSRFortessa is maintained via an annual service contract.

Luminex MAGPIX

The MAGPIX is a versatile multiplexing platform capable of performing qualitative and quantitative magnetic bead-based analysis of proteins and nucleic acids in a variety of sample matrices. This system can perform up to 50 tests in a single reaction volume, greatly reducing sample input, reagents, and labor. The MAGPIX system features an innovative design based on LED excitation and CCD imaging technology that allows for a compact, robust system. MAGPIX is supported by a broad menu of commercially available assay kits, as well as reagents to build your own assays. Data Acquisition and Analysis for Luminex Instrumentation: xPONENT 4.3 is a modular, flexible, software package for control of the MAGPIX. The software’s graphical user interface follows the typical assay workflow but can be customized to meet the needs of a specific laboratory. The MAGPIX is maintained via an annual service contract.

Isoplexis IsoLight Instrumentation

The Duke Immune Profiling Core has an innovative new assay platform to serve the Duke medical research enterprise. These chip-based, highly multiplexed, and single-cell proteomics platforms developed by Isoplexis Inc. are relatively new to the Duke research community and have broad applicability across a wide variety of research disciplines such as novel cellular immunotherapies, solid organ transplantation, pre- and post-medical/surgical interventions, and general immunologic monitoring, to name a few. The key component of this new platform is the IsoLight instrumentation that is in place in DIPC. The Isolight is maintained via an annual service contract. Further details regarding the capabilities of the platform can be gleaned either from the Isoplexis website or by contacting Kent Weinhold or Katelyn Steadman.

Data Management and Archival

Immediately following sample acquisition, all raw data (in the form of FCS files, Isoplexis data files, and/or Luminex csv are exported from the acquisition software to the local computer, then transferred to the local DHTS server in a single DIPC folder location, named Data. From there the data files are both downloaded to a local workstation for subsequent analysis and transferred to long-term storage for archival, with limited access. All investigators have access to download data files as needed from the DHTS Data folder; however, only Core staff and local IT have access to data in the archived folder, where the data are available as read-only and cannot be modified. All archived data are stored on the server permanently. When Core staff performs flow cytometry analysis for clinical trials, we maintain a folder for each trial; the folder includes two subfolders: Assay Docs (protocol or SOP, assay worksheet Excel document, count sheets, completed assay and count sheets, etc) and Analysis Docs (FlowJo analysis file, a Powerpoint or PDF summary of the analysis, Excel containing data directly exported from FlowJo, and a record of any deviations that occurred during the assay or analysis).

Server

File storage is managed and maintained by Duke Health Technology Services (DHTS). It is an expandable NAS that is enterprise-grade, located in the Duke Fitz East Datacenter. It stores clinical and research information for other Duke departments as well. Called Isilon, this NAS grants us 30 days of retrievable copies of any file or folder. As a result of this limitation and to achieve better data integrity, we have created a policy-based archive folder system that is separate but adjacent to the live data folders that our labs use to store data that will not be changed further. Data inside this archive is set as read-only for each group.

Immune Profile Leadership and Staff

Leadership

Staff

  • Jennifer Enzor, 919-684-3315
  • Katelyn Steadman, 919-684-3542
  • Preksha Patel
  • Erin Murray
  • Victoria Little

Location

Surgical Oncology Research Facility
915 South LaSalle Street Ext. Room 120/122/124 Durham, NC 27710
duke.ipc@dm.duke.edu

Selected Achievements

Representative DIPC Collaborative Publications: 

  • Brown, Landon C., Halabi, S., Somarelli, J., Humeniuk, M., Wu, Y., Oyekunle, T., Howard, L., Huang, J., Anand, M., Davies, C., Patel, P., Staats, J., Weinhold, K.J., Harrison, M.R., Zhang, T., George, D.J., and Armstrong, A.J. A phase 2 trial of avelumab in men with aggressive-variant or neuroendocrine prostate cancer. Prostate Cancer and Prostatic Diseases 25(4):762-769, 2022. PMCID: PMC8933335.
  • Lusk, J.B., Quinones, Q.J., Staats, J.S., Weinhold, K.J., Grossi, P.M., Laskowitz, D.T., and James, M.L. Coupling hematoma evacuation with immune profiling for analysis of neuroinflammation after primary intracerebral hemorrhage: a pilot study. World Neurosurg. 2022 May;161:162-168 PMCID:PMID:35217228.
  • Li, Y., Yi, J.S., Russo, M.., Rosa-Bray, M., Weinhold. K.J., and Guptill, J.T. Normative dataset for plasma cytokines in healthy human adults. Data Brief 2021 Feb. 9;35:106857. PMC7900339
  • Salama, A.K.S., Palta, M., Rushing, C.N., Selim, M.A., Linnet, K.N., Czito, B.G., Yoo, D.S., Hanks, B.A., Beasley, G.M., Mosca, P., Dumbauld, C., Steadman, K.N., Yi, J.S., Weinhold, K.J., Tyler, D.S., Lee, W.T., and Brizel, D.M. Ipilimumab and radiation in patients with high risk resected or regionally advanced melanoma. Clin. Cancer Res. 1 March, 2021 27(5):1287-1295. PMID:33172894.
  • Zhang, T., Harrison, M.R., O’Donnell, P.H., Ajjai, A., Hahn, N.M., Appleman, L.J., Cetnar, J., Burke, J.M., Fleming, M., Miloswsky. M., Mortazavi, A., Shore, N., Sonapavde, G., Schmidt, E., Bitman, B., Munugalavadla, V., Izumi, P., Patel, P., Staats, J., Chan, C., Weinhold, K.J.*and George, D.J.,*senior co-authors. Cancer Oct.15, 2020 126(20):4485-4497. PMC7590121.
  • Healy, Z.R., Weinhold, K.J., and Murdoch D.M. Transcriptional profiling of CD8+ CMV-specific T cell functional subsets obtained using a method for isolating high-quality RNA from fixed and permeabilized cells. Frontiers in Immunology 11:1859, Sep. 2, 2020. PMC7492549.
  • Yi, J.S., Rosa-Bray, M., Staats, J., Zakroysky, P., Chan, C., Russo, M., Dumbauld, C., White, S., Gierman, T., Weinhold, K.J., and Guptill, J.T. Establishment of normative ranges of the healthy immune system with comprehensive polychromatic flow cytometry profiling. PLoS One 14(12):e0225512, Dec.11, 2019. PMC6905525.
  • Berger, M., Murdoch, D., Staats, J., Chan, C., Thomas J., Garrigues, G., Browndyke, J., Cooter, M., Quinones, Q., Matthew, J., and Weinhold, K.J. Flow cytometry characterization of cerebrospinal fluid monocytes in patients with postoperative cognitive dysfunction (POCD): A pilot study. Anesthesia & Analgesia May 3, 2019 doi: 10.1213/ANE. PMC6800758.
  • Weinhold, K.J., Bukowski, J.F., Brennan, T.V., Noveck, R.J., Staats, J.S., Lin, L., Stempora, L., Hammond, C., Wouters, A., Mojcik, C.F., Cheng, J., Collinge, M., Jesson, M.I., Hazra, A., Biswas, P., Lan, S., Clark, J.D., and Hodge, J.A. Reversibility of peripheral blood leukocyte phenotypic and functional changes after exposure to and withdrawal from tofacitinib, a Janus kinase inhibitor, in healthy volunteers. Clin. Immunology 191:10-20, June 19, 2018. PMC6036921.
  • Yi, J.S., Ready, N., Healy, P., Dumbauld, C., Berry, M., Shoemaker, D., Clarke, J., Crawford, J., Tong, B.C., Harpole, D., D’Amico, T.A., McSherry, F., Dunphy, F., McCall, S.J., Christensen, J.D., Wang, X, and Weinhold, K.J. Immune activation in early stage non-small cell lung cancer patients receiving neoadjuvant chemotherapy plus ipilimumab. Clin. Cancer Res. 23(24):7474-7482, 2017. PMC5732888.
  • Snyder, L.D., Chan, C., Kwon, D., Yi, J.S., Martissa, J.A., Finlen Copeland, C.A., Osborne, R.J., Sparks, S.D., Palmer, S.M., and Weinhold, K.J.  Polyfunctional T cell signatures to predict protection from cytomegalovirus after lung transplant. Am. J. Respir. Crit. Care. Med. 193 (1): 78-85, Jan 1, 2016. PMC4731614.
  • White, S., Laske, K., Welters, M.J.P., Bidmon, N., van der Burg, S.H., Britten, C.M., Enzor, J., Weinhold, K.J., Gouttefangeas, C., and Chan, C. Managing multi-center flow cytometry data for immune monitoring. Cancer Informatics 13(S7):111-122, 2014.  PMID:26085786
  • Staats, J.S., Enzor, J.H., Sanchez, A.M., Roundtree, W., Guar, A., Jaimes, M., Denny, T.N., and Weinhold, K.J. Toward a comprehensive external quality assurance program for polychromatic flow cytometry. J. Immunol. Methods, 409:44-53, 2014. PMC4138230
  • Richards, A.J., Staats, J., Enzor, J., McKinnon, K., Frelinger, J., Denny, T.N., Weinhold, K.J., and C. Chan. Setting objective thresholds for rare event detection in flow cytometry. J. Immunol. Methods, 409:54-61, 2014. PMC4138273.
  • McNeil, L.K., Price, L., Britten, C.M., Jaimes, M., Maecker, H., Odunsi, K., Matsuzaki, J., Staats, J.S., Thorpe, J., Yuan, J., and Janetzki, S. International consortia proficiency panel conducted by the cancer immunotherapy consortium (CIC/CRI). Cytometry, Part A 83A:728-738, 2013. PMC4443815.
  • Murdoch, D.M., Staats, J.S., and Weinhold, K.J. OMIP-006: Phenotypic subset analysis of T regulatory cells via polychromatic flow cytometry.  Cytometry A 81(4):281-283, 2012. PMC3352600.
  • Snyder, L.D., Medinas, R., Chan, C., Sparks, S., Davis, W.A., Palmer, S.M., and Weinhold, K.J.  Polyfunctional cytomegalovirus-specific immunity in lung transplant recipients receiving valganciclovir.  Am. J. Transplant. 11:1-8, 2011. PMC3044779.
  • Maecker, H.T., McCoy, J.P. Jr., Amos, A., Elliott, J., Gaigalas, A., Wang, L., Aranda, R., Banchereau, J., Boshoff, C., Braun, J., Korin, Y., Reed, E., Cho, J., Hafler, D., Fathman, C.G., Robinson, W., Denny, T., Weinhold, K., Desai, B., Diamond, B., Gregersen, P., DiMeglio, P., Nestle, F., Peakman, M., Villnova, F., Ferbes, J., Field, E., Kantor, A., Kawabata, T., Komocsar, W., Lotze, M., Nepom, J., Ochs, H., O’Lone, R., Phippard, D., Plevy, S., Rich, S., Roederer, M.m Rostrosen, D., and Yeh, J.H.  A model for harmonizing flow cytometry in clinical trials.  Nat. Immunol. 11(11): 975-078, 2010. PMC3400260.