Kent Weinhold


In addition to their ongoing HIV/AIDS-related research activities, the Weinhold Laboratory is focused on utilizing a comprehensive repertoire of highly standardized and formerly validated assay platforms to profile the human immune system in order to identify immunologic signatures that predict disease outcomes. These ongoing studies span a broad range of highly relevant clinical arenas, including: 1) cancer (non-small cell lung cancer, head and neck cancer, glioblastoma neoforme, ovarian cancer, and prostate cancer), 2) autoimmune diseases (rheumatoid arthritis, systemic lupus erythematosis, multiple sclerosis, and myasthenia gravis), 3) pulmonary disease (idiopathic pulmonary fibrosis), 4) solid organ transplantation (lung, kidney, liver, and heart), and 5) rare diseases (Pompe disease).

Two of the areas that have been especially active over the past few years include the comprehensive immunologic profiling of cancer patients receiving so-called ‘immune checkpoint blockade’ therapies and the search for immune signatures in lung transplant recipients that track with resistance to CMV infection. The laboratory is conducting immune monitoring studies associated with a Phase I trial of Ipilimumab (anti-CTLA-4) in a neoadjuvant setting for the treatment of non-small cell lung cancer (NSCLC). For this trial we are extensively utilized several polychromatic flow cytometry (PFC) platforms to follow activation, maturation, exhaustion, and proliferation patterns within CD4+ and CD8+ subsets of T-cells. We are also utilizing an intracellular cytokine staining (ICS) platform in efforts to detect anti-tumor associated antigen (TAA) responses by CD4+ and CD8+ T cells from peripheral blood mononuclear cells as well as lymphocytes infiltrating the patients’ tumor. These assays are designed to measure antigen-driven intracellular production of IFN-γ, TNF-α, and IL-2, as well as the degranulation marker CD107. This strategy enables us to not only document individual cytokine responses, but to also assess (through Boolean gating) changes in relative polyfunctionality of the responses. We are performing similar immune monitoring of a Phase II trial evaluating nivolumab (anti-PD-1) alone vs. combined nivolumamb + ipilimumab vs. avastin (bevacizamab) alone in patients with glioblastomas. In both studies, we are seeking to identify pharmacodynamics markers and immune correlates predictive of clinical responses. In recently completed studies in a cohort of lung transplant recipients, we identified specific polyfunctional signatures in CD4+ and CD8+ subsets against CMV pp65 and IE-1 antigens that tracked with resistance to CMV infection (manuscript in preparation). These findings now serve as the basis for a Phase I clinical trial to compare conventional 6-month chemoprophylaxis in lung transplant recipients versus a regimen dictated by the presence or absence of the predictive signatures. This trial is the principal component of a recently awarded Clinical Trials in Organ Transplantation or CTOT award made from the NIH to Duke (Scott Palmer, PI). Ongoing studies will test the hypothesis that these signatures that have been validated in lung transplant recipients will also predict resistance to CMV infection in the context of other solid organ transplants such as kidney, liver, and heart.

Future studies will also attempt to identify predictive signatures for resistance to BK polyomavirus, the cause of graft threatening nephritis in kidney transplant recipients and cystitis in bone marrow transplant recipients. Other human diseases that are presently being subjected to comprehensive immune profiling by the laboratory include idiopathic pulmonary fibrosis (IPF), myasthenia gravis, multiple sclerosis, systemic lupus erythematosus, acute coronary syndrome, and Pompe disease



Recent publications

Zidar, D.A., Mudd, J.C., Juchnowski, S., Lopes, J.P., Sparks, S., Park, S.S., Ishikawa, M., Osborne, R., Washam, J.B., Chan, C., Funderburg, N.T., Owoyele, A., Alaiti, M.A., Mayuga, M., Orringer, C., Costa, M.A., Simon, D.I., Tatsuoka, C., Califf, R.M., Newby, L.K., Lederman, M.M., and Weinhold, K.J.  Altered maturation status and possible immune exhaustion of CD8 T lymphocytes in the peripheral blood of patients presenting with acute coronary syndromes. Arterioscler., Thromb., and Vasc. Biol. 36(2): 389-397, Feb. 2016 PMID: 26663396

Yi, J.S., Russo, M.A., Weinhold, K.J., and Guptill, J.T. Adaptive immune response to therapy in HMGCR autoantibody myopathy. Muscle Nerve.  53(2): 313-317, Feb. 2016. PMID: 26492512          

Snyder, L.D., Chan, C., Kwon, D., Yi J.S., Martissa, J.A., Copeland, C.A.F., Osborne, R.J., Sparks, S.D., Palmer, S.M., Weinhold, K.J.  Polyfunctional T cell responses predict protection from cytomegalovirus after lung transplant.  Am J Respir Crit Care Med. 193 (1): 78-85, Jan.1, 2016 [PMID 26372850]

Guptill, J.T., Yi, J.S., Sanders, D.B., Guidon, A.C., Juel, V.C., Massey, J.M., Howard, J.F., Jr., Scuderi, F., Bartoccioni, E., Evoli, A. and Weinhold, K.J. “Characterization of B cells in muscle-specific kinase antibody myasthenia gravis. Neurol Neuroimmunol Neuroinflamm. 2015 Feb 26;2(2) e77. [PMID 25745635]

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. [PMID 24968072]

Nair, S.K., Tomaras, G.D., Sales A.P., Boczkowski, D., Chan, C., Plonk, K., Dannull, J., Pruitt, S.K., and Weinhold, K.J. “High-throughput identification and dendritic cell-based functional validation of MHC class I-restricted Mycobacterium tuberculosis epitopes.” Scientific Reports. Apr. 23, 2014. [PMID 24755960]

Yi, J.S., Guidon, A., Sparks, S., Osborne, R., Juel, V.C., Massey, J.M., Sanders, D.B., Weinhold, K.J., and Guptill, J.T. “Characterization of CD4 and CD8 T cell responses in MuSK myasthenia gravis.” Journal of Autoimmunity. 2013 Dec27 doi:1016/j.jaunt2013. 12.005. [PMID: 24378287]


Professor in Surgery

Surgery, Surgical Sciences
School of Medicine

Joseph W. and Dorothy W. Beard Distinguished Professor of Experimental Surgery

Surgery, Surgical Sciences
School of Medicine

Chief, Division of Surgical Sciences

Surgery, Surgical Sciences
School of Medicine

Professor of Immunology

School of Medicine

Professor in Pathology

School of Medicine

Member of the Duke Human Vaccine Institute

Duke Human Vaccine Institute
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine


Ph.D. 1979

Thomas Jefferson University


UNC - Duke Immunotherapy Training Program

Administered By
Medicine, Hematologic Malignancies and Cellular Therapy
Awarded By
University of North Carolina - Chapel Hill
Start Date
End Date

Duke KURe Program

Administered By
Obstetrics and Gynecology, Urogynecology
Awarded By
National Institutes of Health
Start Date
End Date

Interdisciplinary Training Program in Lung Disease

Administered By
Medicine, Pulmonary, Allergy, and Critical Care Medicine
Awarded By
National Institutes of Health
Start Date
End Date

Institutional Training Grant in Pediatric Infectious Disease

Administered By
Pediatrics, Infectious Diseases
Awarded By
National Institutes of Health
Start Date
End Date

Targeting EGFRvIII in Brain Tumors with Bispecific Antibodies

Administered By
School of Medicine
Awarded By
National Institutes of Health
Start Date
End Date


FlowKit: A Python Toolkit for Integrated Manual and Automated Cytometry Analysis Workflows.

An important challenge for primary or secondary analysis of cytometry data is how to facilitate productive collaboration between domain and quantitative experts. Domain experts in cytometry laboratories and core facilities increasingly recognize the need for automated workflows in the face of increasing data complexity, but by and large, still conduct all analysis using traditional applications, predominantly FlowJo. To a large extent, this cuts domain experts off from the rapidly growing library of Single Cell Data Science algorithms available, curtailing the potential contributions of these experts to the validation and interpretation of results. To address this challenge, we developed FlowKit, a Gating-ML 2.0-compliant Python package that can read and write FCS files and FlowJo workspaces. We present examples of the use of FlowKit for constructing reporting and analysis workflows, including round-tripping results to and from FlowJo for joint analysis by both domain and quantitative experts.
White, S; Quinn, J; Enzor, J; Staats, J; Mosier, SM; Almarode, J; Denny, TN; Weinhold, KJ; Ferrari, G; Chan, C
MLA Citation
White, Scott, et al. “FlowKit: A Python Toolkit for Integrated Manual and Automated Cytometry Analysis Workflows.Front Immunol, vol. 12, 2021, p. 768541. Pubmed, doi:10.3389/fimmu.2021.768541.
Published In
Frontiers in Immunology
Published Date
Start Page

Coupling Hematoma Evacuation with Immune Profiling for Analysis of Neuroinflammation After Primary Intracerebral Hemorrhage: A Pilot Study.

OBJECTIVE: We sought to explore the use and feasibility of an integrated hematoma evacuation/tissue preservation system coupled with immune profiling to assess human ex vivo immune cell populations from brain hematoma samples after intracerebral hemorrhage (ICH). METHODS: In this nonrandomized, noncontrolled pilot/feasibility study of 7 patients with primary supratentorial ICH, a hematoma evacuation device and integrated tissue preservation system were used to obtain hematoma samples during surgical evacuation. Samples were processed, cryopreserved, and analyzed using flow cytometry to determine the relative distribution of immune cell populations compared with peripheral blood mononuclear cells from healthy control subjects. RESULTS: This study demonstrates proof of concept for an integrated hematoma evacuation and sample preservation system to collect human brain hematoma samples for flow cytometry analysis after acute human ICH. In our preliminary analysis, hematoma samples demonstrated a different makeup of white blood cells than peripheral blood from healthy controls. CONCLUSIONS: Flow cytometry analysis of hematoma samples in ICH demonstrates the potential to provide important insights into neuroinflammation associated with ICH.
Lusk, JB; Quinones, QJ; Staats, JS; Weinhold, KJ; Grossi, PM; Nimjee, SM; Laskowitz, DT; James, ML
MLA Citation
Lusk, Jay B., et al. “Coupling Hematoma Evacuation with Immune Profiling for Analysis of Neuroinflammation After Primary Intracerebral Hemorrhage: A Pilot Study.World Neurosurg, vol. 161, May 2022, pp. 162–68. Pubmed, doi:10.1016/j.wneu.2022.02.062.
Published In
World Neurosurg
Published Date
Start Page
End Page

A phase 2 trial of avelumab in men with aggressive-variant or neuroendocrine prostate cancer.

BACKGROUND: Men with progressive neuroendocrine or aggressive-variant metastatic prostate cancer (NEPC/AVPC) have a poor prognosis and limited treatment options, and immunotherapy has not been tested in such patients. METHODS: We conducted an open label single center phase 2 trial (NCT03179410) of men with progressive NEPC/AVPC either defined by histology or AVPC criteria. Avelumab (10 mg/kg every 2 weeks) was administered until progression or unacceptable toxicity. The primary endpoint was overall response rate (ORR). Secondary endpoints included ORR, radiographic progression-free survival (rPFS), overall survival, and safety. Correlative studies included longitudinal peripheral blood immune phenotyping. The study was limited by the small number of patients enrolled and by the early termination due to COVID-19. RESULTS: A total of 15 men with AVPC/NEPC were enrolled. The median age was 71 (range 51-85 years), and men had received a median of two prior therapies (range 1-3). Median PSA was 54 ng/dl (range 0-393), and 73% of men had liver metastasis. The ORR with avelumab in this setting by iRECIST or RECIST 1.1 was 6.7%, including one patient (6.7%) with a complete remission (CR), 20% with stable disease, and 67% with progressive disease. The patient with the CR had an MSH2 somatic mutation and MSI-high NEPC with central nervous system metastases, and his CR remains durable off all therapy for 2 years. The median rPFS was 1.8 months (95% CI 1.6-3.6 months), and median overall survival was 7.4 months (85% CI 2.8-12.6 months). Safety was consistent with the known profile of avelumab. Phenotyping of peripheral immune subsets suggest enhanced CXCR2-dependent myeloid and T-cell responses in this extraordinary responder. CONCLUSIONS: While the study was terminated early due to slow enrollment at the onset of the COVID-19 pandemic and lower than anticipated objective response rate, PD-L1 inhibition with avelumab monotherapy showed poor efficacy in patients with microsatellite stable NEPC/AVPC. Immune profiling revealed enhanced CXCR2 positive immune cell activation in the one extraordinary responder, suggesting potential mechanisms for further immunotherapy development in this population.
Brown, LC; Halabi, S; Somarelli, JA; Humeniuk, M; Wu, Y; Oyekunle, T; Howard, L; Huang, J; Anand, M; Davies, C; Patel, P; Staats, J; Weinhold, KJ; Harrison, MR; Zhang, T; George, DJ; Armstrong, AJ
MLA Citation
Brown, Landon C., et al. “A phase 2 trial of avelumab in men with aggressive-variant or neuroendocrine prostate cancer.Prostate Cancer Prostatic Dis, Mar. 2022. Pubmed, doi:10.1038/s41391-022-00524-7.
Published In
Prostate Cancer Prostatic Dis
Published Date

Wnt activation promotes memory T cell polyfunctionality via epigenetic regulator PRMT1.

T cell polyfunctionality is a hallmark of protective immunity against pathogens and cancer, yet the molecular mechanism governing it remains mostly elusive. We found that canonical Wnt agonists inhibited human memory CD8+ T cell differentiation while simultaneously promoting the generation of highly polyfunctional cells. Downstream effects of Wnt activation persisted after removal of the drug, and T cells remained polyfunctional following subsequent cell division, indicating the effect is epigenetically regulated. Wnt activation induced a gene expression pattern that is enriched with stem cell-specific gene signatures and upregulation of protein arginine methyltransferase 1 (PRMT1), a known epigenetic regulator. PRMT1+CD8+ T cells are associated with enhanced polyfunctionality, especially the ability to produce IL-2. In contrast, inhibition of PRMT1 ameliorated the effects of Wnt on polyfunctionality. Chromatin immunoprecipitation revealed that H4R3me2a, a permissive transcription marker mediated by PRMT1, increased at the IL-2 promoter loci following Wnt activation. In vivo, Wnt-treated T cells exhibited superior polyfunctionality and persistence. When applied to cytomegalovirus (CMV) donor-seropositive, recipient-seronegative patients (D+/R-) lung transplant patient samples, Wnt activation enhanced CMV-specific T cell polyfunctionality, which is important in controlling CMV diseases. These findings reveal a molecular mechanism governing T cell polyfunctionality and identify PRMT1 as a potential target for T cell immunotherapy.
Sung, B-Y; Lin, Y-H; Kong, Q; Shah, PD; Glick Bieler, J; Palmer, S; Weinhold, KJ; Chang, H-R; Huang, H; Avery, RK; Schneck, J; Chiu, Y-L
MLA Citation
Sung, Bo-Yi, et al. “Wnt activation promotes memory T cell polyfunctionality via epigenetic regulator PRMT1.J Clin Invest, vol. 132, no. 2, Jan. 2022. Pubmed, doi:10.1172/JCI140508.
Published In
J Clin Invest
Published Date

Physiological Fitness and the Pathophysiology of Chronic Lymphocytic Leukemia (CLL).

Chronic lymphocytic leukemia (CLL) is associated with physical dysfunction and low overall fitness that predicts poor survival following the commencement of treatment. However, it remains unknown whether higher fitness provides antioncogenic effects. We identified ten fit (CLL-FIT) and ten less fit (CLL-UNFIT) treatment-naïve CLL patients from 144 patients who completed a set of physical fitness and performance tests. Patient plasma was used to determine its effects on an in vitro 5-day growth/viability of three B-cell cell lines (OSU-CLL, Daudi, and Farage). Plasma exosomal miRNA profiles, circulating lipids, lipoproteins, inflammation levels, and immune cell phenotypes were also assessed. CLL-FIT was associated with fewer viable OSU-CLL cells at Day 1 (p = 0.003), Day 4 (p = 0.001), and Day 5 (p = 0.009). No differences between the groups were observed for Daudi and Farage cells. Of 455 distinct exosomal miRNAs identified, 32 miRNAs were significantly different between the groups. Of these, 14 miRNAs had ≤-1 or ≥1 log2 fold differences. CLL-FIT patients had five exosomal miRNAs with lower expression and nine miRNAs with higher expression. CLL-FIT patients had higher HDL cholesterol, lower inflammation, and lower levels of triglyceride components (all p < 0.05). CLL-FIT patients had lower frequencies of low-differentiated NKG2+/CD158a/bneg (p = 0.015 and p = 0.014) and higher frequencies of NKG2Aneg/CD158b+ mature NK cells (p = 0.047). The absolute number of lymphocytes, including CD19+/CD5+ CLL-cells, was similar between the groups (p = 0.359). Higher physical fitness in CLL patients is associated with altered CLL-like cell line growth in vitro and with altered circulating and cellular factors indicative of better immune functions and tumor control.
Sitlinger, A; Deal, MA; Garcia, E; Thompson, DK; Stewart, T; MacDonald, GA; Devos, N; Corcoran, D; Staats, JS; Enzor, J; Weinhold, KJ; Brander, DM; Weinberg, JB; Bartlett, DB
MLA Citation
Sitlinger, Andrea, et al. “Physiological Fitness and the Pathophysiology of Chronic Lymphocytic Leukemia (CLL).Cells, vol. 10, no. 5, May 2021. Pubmed, doi:10.3390/cells10051165.
Published In
Published Date

Research Areas:

AIDS Vaccines
Acquired Immunodeficiency Syndrome
Administration, Oral
African Americans
Age Factors
Amino Acid Sequence
Antibodies, Monoclonal
Antibodies, Viral
Antibody-Dependent Cell Cytotoxicity
Antigen Presentation
Antigen-Presenting Cells
Antigens, CD
Antigens, CD4
Antigens, CD8
Antigens, Differentiation, T-Lymphocyte
Antigens, Neoplasm
Antigens, Surface
Antiretroviral Therapy, Highly Active
Antiviral Agents
Automation, Laboratory
B-Cell Activating Factor
Binding Sites, Antibody
Binding, Competitive
Bone Marrow
Bone Marrow Cells
CD4 Antigens
CD4-CD8 Ratio
CD4-Positive T-Lymphocytes
CD8-Positive T-Lymphocytes
Canarypox virus
Carbohydrate Metabolism
Case-Control Studies
Cell Count
Cell Proliferation
Cell Separation
Cell Transformation, Neoplastic
Chemotherapy, Cancer, Regional Perfusion
Child, Preschool
Clinical Trials as Topic
Clone Cells
Coculture Techniques
Cohort Studies
Combined Modality Therapy
Connective Tissue
Cross Reactions
Cytomegalovirus Infections
Cytopathogenic Effect, Viral
Cytotoxicity Tests, Immunologic
Cytotoxicity, Immunologic
DNA Primers
DNA, Viral
DNA-Directed DNA Polymerase
Dendritic Cells
Disease Progression
Dose-Response Relationship, Immunologic
Double-Blind Method
Drug Resistance, Viral
Enzyme-Linked Immunosorbent Assay
Epitope Mapping
Epitopes, T-Lymphocyte
European Continental Ancestry Group
Evolution, Molecular
Fas Ligand Protein
Flow Cytometry
Fluorescent Antibody Technique
Follow-Up Studies
Freund's Adjuvant
Gene Expression
Gene Expression Regulation
Gene Products, gag
Gene Rearrangement, T-Lymphocyte
Genes, Viral
Genome, Viral
Guideline Adherence
HIV Antibodies
HIV Antigens
HIV Envelope Protein gp120
HIV Envelope Protein gp160
HIV Envelope Protein gp41
HIV Infections
HIV Long Terminal Repeat
HIV Reverse Transcriptase
HIV Seronegativity
HIV Seropositivity
HLA-B27 Antigen
Hematopoietic Stem Cells
Hemophilia A
High-Throughput Screening Assays
Histocompatibility Antigens Class I
Immune Evasion
Immune Sera
Immune System
Immune Tolerance
Immunity, Innate
Immunity, Mucosal
Immunization Schedule
Immunization, Secondary
Immunoglobulin A
Immunoglobulin G
Immunoglobulin M
Immunologic Deficiency Syndromes
Immunologic Memory
Immunosuppressive Agents
In Situ Hybridization
Inflammation Mediators
Infusions, Parenteral
International Cooperation
Intestinal Absorption
Killer Cells, Natural
Leukocyte Count
Leukocytes, Mononuclear
Lymphocyte Activation
Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
Lymphocyte Subsets
Major Histocompatibility Complex
Mice, Inbred C3H
Middle Aged
Molecular Sequence Data
Myasthenia Gravis
Neisseria gonorrhoeae
Neoplasm Proteins
Neoplasm Transplantation
Neutralization Tests
Normal Distribution
Observer Variation
Pan troglodytes
Pancreas Transplantation
Peptide Fragments
Peptide Mapping
Pilot Projects
Pneumonia, Pneumocystis
Polymerase Chain Reaction
Program Development
Program Evaluation
Prospective Studies
Protein Binding
Protein Processing, Post-Translational
Protein-Tyrosine Kinases
Proto-Oncogene Proteins
Proto-Oncogene Proteins c-fyn
Quality Control
Quality Improvement
Quality Indicators, Health Care
RNA-Directed DNA Polymerase
Rabies Vaccines
Receptors, Antigen, T-Cell
Receptors, Antigen, T-Cell, gamma-delta
Receptors, Fc
Receptors, IgG
Receptors, Tumor Necrosis Factor, Type II
Recombinant Proteins
Reference Values
Regression Analysis
Remission Induction
Reproducibility of Results
Retrospective Studies
Retroviridae Infections
Retroviridae Proteins
Reverse Transcriptase Inhibitors
Risk Factors
SAIDS Vaccines
Sarcoma, Kaposi
Sequence Analysis, Protein
Sequence Homology, Amino Acid
Sex Factors
Sexual Behavior
Sexually Transmitted Diseases
Signal Transduction
Simian Acquired Immunodeficiency Syndrome
Simian immunodeficiency virus
Skin Neoplasms
Specimen Handling
Stem Cells
Substrate Specificity
Suppressor Factors, Immunologic
Surface Properties
Survival Rate
T-Cell Antigen Receptor Specificity
T-Lymphocyte Subsets
T-Lymphocytes, Cytotoxic
T-Lymphocytes, Helper-Inducer
TNF-Related Apoptosis-Inducing Ligand
Thymidine Monophosphate
Thymine Nucleotides
Tissue Distribution
Transcription, Genetic
Transduction, Genetic
Transplantation, Homologous
Treatment Outcome
Trinidad and Tobago
Tumor Cells, Cultured
Tumor Virus Infections
United States
Vaccines, DNA
Vaccines, Subunit
Vaccines, Synthetic
Vaccinia virus
Viral Envelope Proteins
Viral Load
Viral Proteins
Virus Replication
Young Adult
ZAP-70 Protein-Tyrosine Kinase
env Gene Products, Human Immunodeficiency Virus
src-Family Kinases