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Article ; Online: Tracking of activated cTfh cells following sequential influenza vaccinations reveals transcriptional profile of clonotypes driving a vaccine-induced immune response.

Currenti, Jennifer / Simmons, Joshua / Oakes, Jared / Gaudieri, Silvana / Warren, Christian M / Gangula, Rama / Alves, Eric / Ram, Ramesh / Leary, Shay / Armitage, Jesse D / Smith, Rita M / Chopra, Abha / Halasa, Natasha B / Pilkinton, Mark A / Kalams, Spyros A

Frontiers in immunology

2023 Volume 14, Page(s) 1133781

Abstract: Introduction: A vaccine against influenza is available seasonally but is not 100% effective. A predictor of successful seroconversion in adults is an increase in activated circulating T follicular helper (cTfh) cells after vaccination. However, the ... ...

Abstract	Introduction: A vaccine against influenza is available seasonally but is not 100% effective. A predictor of successful seroconversion in adults is an increase in activated circulating T follicular helper (cTfh) cells after vaccination. However, the impact of repeated annual vaccinations on long-term protection and seasonal vaccine efficacy remains unclear. Methods: In this study, we examined the T cell receptor (TCR) repertoire and transcriptional profile of vaccine-induced expanded cTfh cells in individuals who received sequential seasonal influenza vaccines. We measured the magnitude of cTfh and plasmablast cell activation from day 0 (d0) to d7 post-vaccination as an indicator of a vaccine response. To assess TCR diversity and T cell expansion we sorted activated and resting cTfh cells at d0 and d7 post-vaccination and performed TCR sequencing. We also single cell sorted activated and resting cTfh cells for TCR analysis and transcriptome sequencing. Results and discussion: The percent of activated cTfh cells significantly increased from d0 to d7 in each of the 2016-17 (p < 0.0001) and 2017-18 (p = 0.015) vaccine seasons with the magnitude of cTfh activation increase positively correlated with the frequency of circulating plasmablast cells in the 2016-17 (p = 0.0001) and 2017-18 (p = 0.003) seasons. At d7 post-vaccination, higher magnitudes of cTfh activation were associated with increased clonality of cTfh TCR repertoire. The TCRs from vaccine-expanded clonotypes were identified and tracked longitudinally with several TCRs found to be present in both years. The transcriptomic profile of these expanded cTfh cells at the single cell level demonstrated overrepresentation of transcripts of genes involved in the type-I interferon pathway, pathways involved in gene expression, and antigen presentation and recognition. These results identify the expansion and transcriptomic profile of vaccine-induced cTfh cells important for B cell help.
MeSH term(s)	Adult ; Humans ; Influenza, Human/prevention & control ; B-Lymphocytes ; Vaccination ; Influenza Vaccines ; Immunity
Chemical Substances	Influenza Vaccines
Language	English
Publishing date	2023-03-29
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2023.1133781
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Article ; Online: In chronic infection, HIV gag-specific CD4+ T cell receptor diversity is higher than CD8+ T cell receptor diversity and is associated with less HIV quasispecies diversity.

Pilkinton, Mark A / McDonnell, Wyatt J / Barnett, Louise / Chopra, Abha / Gangula, Rama / White, Katie D / Leary, Shay / Currenti, Jennifer / Gaudieri, Silvana / Mallal, Simon A / Kalams, Spyros A

Journal of virology

2021 Volume 95, Issue 8

Abstract: Cellular immune responses to Gag correlate with improved HIV viral control. The full extent of cellular immune responses comprise both the number of epitopes recognized by CD4+ and CD8+ T cells, as well as the diversity of the T cell receptor (TCR) ... ...

Abstract	Cellular immune responses to Gag correlate with improved HIV viral control. The full extent of cellular immune responses comprise both the number of epitopes recognized by CD4+ and CD8+ T cells, as well as the diversity of the T cell receptor (TCR) repertoire directed against each epitope. The optimal diversity of the responsive TCR repertoire is unclear. Therefore, we evaluated the TCR diversity of CD4+ and CD8+ T cells responding to HIV-1 Gag to determine if TCR diversity correlates with clinical or virologic metrics. Previous studies of TCR repertoires have been limited primarily to CD8+ T cell responses directed against a small number of well-characterized T cell epitopes restricted by specific human leucocyte antigens. We stimulated peripheral blood mononuclear cells from 21chronic HIV-infected individuals overnight with a pool of HIV-1 Gag peptides, followed by sorting of activated CD4+ and CD8+ T cells and TCR deep sequencing. We found Gag-reactive CD8+ T cells to be more oligoclonal, with a few dominant TCRs comprising the bulk of the repertoire, compared to the highly diverse TCR repertoires of Gag-reactive CD4+ T cells. HIV viral sequencing of the same donors revealed that high CD4+ T cell TCR diversity was strongly associated with lower HIV Gag genetic diversity. We conclude that the TCR repertoire of Gag-reactive CD4+ T helper cells display substantial diversity without a clearly dominant circulating TCR clonotype, in contrast to a hierarchy of dominant TCR clonotypes in the Gag-reactive CD8+ T cells, and may serve to limit HIV diversity during chronic infection.
Language	English
Publishing date	2021-02-03
Publishing country	United States
Document type	Journal Article
ZDB-ID	80174-4
ISSN	1098-5514 ; 0022-538X
ISSN (online)	1098-5514
ISSN	0022-538X
DOI	10.1128/JVI.02380-20
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Article ; Online: Group 2 Innate Lymphoid Cells Coordinate Damage Response in the Stomach.

Meyer, Anne R / Engevik, Amy C / Madorsky, Toni / Belmont, Erika / Stier, Matthew T / Norlander, Allison E / Pilkinton, Mark A / McDonnell, Wyatt J / Weis, Jared A / Jang, Bogun / Mallal, Simon A / Peebles, R Stokes / Goldenring, James R

Gastroenterology

2020 Volume 159, Issue 6, Page(s) 2077–2091.e8

Abstract: Background & aims: Severe injury to the lining of the stomach leads to changes in the epithelium (reprogramming) that protect and promote repair of the tissue, including development of spasmolytic polypeptide-expressing metaplasia (SPEM) and tuft and ... ...

Abstract	Background & aims: Severe injury to the lining of the stomach leads to changes in the epithelium (reprogramming) that protect and promote repair of the tissue, including development of spasmolytic polypeptide-expressing metaplasia (SPEM) and tuft and foveolar cell hyperplasia. Acute gastric damage elicits a type-2 inflammatory response that includes production of type-2 cytokines and infiltration by eosinophils and alternatively activated macrophages. Stomachs of mice that lack interleukin 33 (IL33) or interleukin 13 (IL13) did not undergo epithelial reprogramming after drug-induced injury. We investigated the role of group 2 innate lymphoid cells (ILC2s) in gastric epithelial repair. Methods: Acute gastric injury was induced in C57BL/6J mice (wild-type and RAG1 knockout) by administration of L635. We isolated ILC2s by flow cytometry from stomachs of mice that were and were not given L635 and performed single-cell RNA sequencing. ILC2s were depleted from wild-type and RAG1-knockout mice by administration of anti-CD90.2. We assessed gastric cell lineages, markers of metaplasia, inflammation, and proliferation. Gastric tissue microarrays from patients with gastric adenocarcinoma were analyzed by immunostaining. Results: There was a significant increase in the number of GATA3-positive ILC2s in stomach tissues from wild-type mice after L635-induced damage, but not in stomach tissues from IL33-knockout mice. We characterized a marker signature of gastric mucosal ILC2s and identified a transcription profile of metaplasia-associated ILC2s, which included changes in expression of Il5, Il13, Csf2, Pd1, and Ramp3; these changes were validated by quantitative polymerase chain reaction and immunocytochemistry. Depletion of ILC2s from mice blocked development of metaplasia after L635-induced injury in wild-type and RAG1-knockout mice and prevented foveolar and tuft cell hyperplasia and infiltration or activation of macrophages after injury. Numbers of ILC2s were increased in stomach tissues from patients with SPEM compared with patients with normal corpus mucosa. Conclusions: In analyses of stomach tissues from mice with gastric tissue damage and patients with SPEM, we found evidence of type 2 inflammation and increased numbers of ILC2s. Our results suggest that ILC2s coordinate the metaplastic response to severe gastric injury.
MeSH term(s)	Animals ; Disease Models, Animal ; Gastric Mucosa/drug effects ; Gastric Mucosa/immunology ; Gastric Mucosa/pathology ; Humans ; Immunity, Innate ; Interleukin-33/genetics ; Lymphocyte Subsets/immunology ; Metaplasia/chemically induced ; Metaplasia/genetics ; Metaplasia/immunology ; Mice ; Mice, Knockout
Chemical Substances	Il33 protein, mouse ; Interleukin-33
Language	English
Publishing date	2020-09-04
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	80112-4
ISSN	1528-0012 ; 0016-5085
ISSN (online)	1528-0012
ISSN	0016-5085
DOI	10.1053/j.gastro.2020.08.051
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Article: Greater activation of peripheral T follicular helper cells following high dose influenza vaccine in older adults forecasts seroconversion

Pilkinton, Mark A / Christian M. Warren / H. Keipp Talbot / Katherine J. Nicholas / Rita M. Smith / Sandra M. Yoder / Spyros A. Kalams

Vaccine. 2017 Jan. 05, v. 35, no. 2

2017

Abstract: Influenza related morbidity and mortality disproportionately impacts older adults. The serologic response to vaccine is diminished in older adults; however, high dose inactivated influenza vaccine (HD IIV) has shown improved rates of seroconversion ... ...

Abstract	Influenza related morbidity and mortality disproportionately impacts older adults. The serologic response to vaccine is diminished in older adults; however, high dose inactivated influenza vaccine (HD IIV) has shown improved rates of seroconversion compared to standard dose (SD IIV). We hypothesize this may be due to the superior ability of high dose vaccine to activate T follicular helper (Tfh) cells and provide B cell dependent T cell help.We measured peripheral Tfh (pTfh) activation in 50 community dwelling adults 65years or older who were randomly assigned to receive either the HD IIV or SD IIV.The HD vaccination elicited significantly higher levels of ICOS expression on pTfh cells, at day 7 compared to SD vaccination (p=0.02). The magnitude of the increase in ICOS+ pTfh cells from baseline to day 7 was predictive of seroconversion for both influenza A and B vaccination.Strong Tfh activation in response to influenza vaccination forecasts successful seroconversion in older adults, and HD IIV elicits greater Tfh activation than SD IIV. Future vaccine studies should focus on ways to further optimize the Tfh response.
Keywords	elderly ; influenza ; morbidity ; mortality ; seroconversion ; T-lymphocytes ; vaccination ; vaccines
Language	English
Dates of publication	2017-0105
Size	p. 329-336.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2016.11.059
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Article ; Online: Greater activation of peripheral T follicular helper cells following high dose influenza vaccine in older adults forecasts seroconversion.

Pilkinton, Mark A / Nicholas, Katherine J / Warren, Christian M / Smith, Rita M / Yoder, Sandra M / Talbot, H Keipp / Kalams, Spyros A

Vaccine

2016 Volume 35, Issue 2, Page(s) 329–336

Abstract: Background: Influenza related morbidity and mortality disproportionately impacts older adults. The serologic response to vaccine is diminished in older adults; however, high dose inactivated influenza vaccine (HD IIV) has shown improved rates of ... ...

Abstract	Background: Influenza related morbidity and mortality disproportionately impacts older adults. The serologic response to vaccine is diminished in older adults; however, high dose inactivated influenza vaccine (HD IIV) has shown improved rates of seroconversion compared to standard dose (SD IIV). We hypothesize this may be due to the superior ability of high dose vaccine to activate T follicular helper (Tfh) cells and provide B cell dependent T cell help. Methods: We measured peripheral Tfh (pTfh) activation in 50 community dwelling adults 65years or older who were randomly assigned to receive either the HD IIV or SD IIV. Results: The HD vaccination elicited significantly higher levels of ICOS expression on pTfh cells, at day 7 compared to SD vaccination (p=0.02). The magnitude of the increase in ICOS+ pTfh cells from baseline to day 7 was predictive of seroconversion for both influenza A and B vaccination. Conclusion: Strong Tfh activation in response to influenza vaccination forecasts successful seroconversion in older adults, and HD IIV elicits greater Tfh activation than SD IIV. Future vaccine studies should focus on ways to further optimize the Tfh response.
MeSH term(s)	Aged ; Aged, 80 and over ; Female ; Humans ; Influenza Vaccines/administration & dosage ; Influenza Vaccines/immunology ; Male ; Seroconversion ; T-Lymphocytes, Helper-Inducer/immunology
Chemical Substances	Influenza Vaccines
Language	English
Publishing date	2016-12-03
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2016.11.059
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Article ; Online: Cross-Reactivity to Mutated Viral Immune Targets Can Influence CD8

Frontiers in immunology

2021 Volume 12, Page(s) 746986

Abstract: Loss of T cell immunogenicity due to mutations in virally encoded epitopes is a well-described adaptation strategy to limit host anti-viral immunity. Another described, but less understood, adaptation strategy involves the selection of mutations within ... ...

Abstract	Loss of T cell immunogenicity due to mutations in virally encoded epitopes is a well-described adaptation strategy to limit host anti-viral immunity. Another described, but less understood, adaptation strategy involves the selection of mutations within epitopes that retain immune recognition, suggesting a benefit for the virus despite continued immune pressure (termed non-classical adaptation). To understand this adaptation strategy, we utilized a single cell transcriptomic approach to identify features of the HIV-specific CD8
MeSH term(s)	Adaptation, Physiological/immunology ; Adult ; CD8-Positive T-Lymphocytes/immunology ; Cross Reactions/immunology ; Epitopes, T-Lymphocyte/immunology ; Female ; HIV/immunology ; HIV Infections/immunology ; Humans ; Male ; Middle Aged
Chemical Substances	Epitopes, T-Lymphocyte
Language	English
Publishing date	2021-10-26
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2021.746986
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Article ; Online: High CD8 T-Cell Receptor Clonality and Altered CDR3 Properties Are Associated With Elevated Isolevuglandins in Adipose Tissue During Diet-Induced Obesity.

McDonnell, Wyatt J / Koethe, John R / Mallal, Simon A / Pilkinton, Mark A / Kirabo, Annet / Ameka, Magdalene K / Cottam, Matthew A / Hasty, Alyssa H / Kennedy, Arion J

Diabetes

2018 Volume 67, Issue 11, Page(s) 2361–2376

Abstract: Adipose tissue (AT) ... ...

Abstract	Adipose tissue (AT) CD4
MeSH term(s)	Adipose Tissue/immunology ; Adipose Tissue/metabolism ; Animals ; CD8-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/metabolism ; Complementarity Determining Regions/metabolism ; Diet, High-Fat ; Glucose Tolerance Test ; Insulin Resistance ; Liver/immunology ; Liver/metabolism ; Male ; Mice ; Obesity/immunology ; Obesity/metabolism ; Prostaglandins/metabolism
Chemical Substances	Complementarity Determining Regions ; Prostaglandins
Language	English
Publishing date	2018-09-04
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	80085-5
ISSN	1939-327X ; 0012-1797
ISSN (online)	1939-327X
ISSN	0012-1797
DOI	10.2337/db18-0040
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Article ; Online: Adipose Tissue in Persons With HIV Is Enriched for CD4

Wanjalla, Celestine N / McDonnell, Wyatt J / Barnett, Louise / Simmons, Joshua D / Furch, Briana D / Lima, Morgan C / Woodward, Beverly O / Fan, Run / Fei, Ye / Baker, Paxton G / Ram, Ramesh / Pilkinton, Mark A / Mashayekhi, Mona / Brown, Nancy J / Mallal, Simon A / Kalams, Spyros A / Koethe, John R

Frontiers in immunology

2019 Volume 10, Page(s) 408

Abstract: Chronic T cell activation and accelerated immune senescence are hallmarks of HIV infection, which may contribute to the increased risk of cardiometabolic diseases in people living with HIV (PLWH). T lymphocytes play a central role in modulating adipose ... ...

Abstract	Chronic T cell activation and accelerated immune senescence are hallmarks of HIV infection, which may contribute to the increased risk of cardiometabolic diseases in people living with HIV (PLWH). T lymphocytes play a central role in modulating adipose tissue inflammation and, by extension, adipocyte energy storage and release. Here, we assessed the CD4
MeSH term(s)	Adipose Tissue/immunology ; Adult ; Antigens, CD/immunology ; Antigens, Differentiation, T-Lymphocyte/immunology ; CD4-Positive T-Lymphocytes/immunology ; CD57 Antigens/immunology ; CX3C Chemokine Receptor 1/immunology ; Female ; Glucose Intolerance/immunology ; HIV Infections/immunology ; Humans ; Immunologic Memory/immunology ; Lectins, C-Type/immunology ; Male ; Middle Aged ; Receptors, G-Protein-Coupled/immunology ; T-Lymphocyte Subsets/immunology
Chemical Substances	ADGRG1 protein, human ; Antigens, CD ; Antigens, Differentiation, T-Lymphocyte ; CD57 Antigens ; CD69 antigen ; CX3C Chemokine Receptor 1 ; CX3CR1 protein, human ; Lectins, C-Type ; Receptors, G-Protein-Coupled
Language	English
Publishing date	2019-03-19
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2019.00408
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Article ; Online: Single-cell analysis shows that adipose tissue of persons with both HIV and diabetes is enriched for clonal, cytotoxic, and CMV-specific CD4+ T cells.

Wanjalla, Celestine N / McDonnell, Wyatt J / Ram, Ramesh / Chopra, Abha / Gangula, Rama / Leary, Shay / Mashayekhi, Mona / Simmons, Joshua D / Warren, Christian M / Bailin, Samuel / Gabriel, Curtis L / Guo, Liang / Furch, Briana D / Lima, Morgan C / Woodward, Beverly O / Hannah, LaToya / Pilkinton, Mark A / Fuller, Daniela T / Kawai, Kenji /

Virmani, Renu / Finn, Aloke V / Hasty, Alyssa H / Mallal, Simon A / Kalams, Spyros A / Koethe, John R

Cell reports. Medicine

2021 Volume 2, Issue 2, Page(s) 100205

Abstract: Persons with HIV are at increased risk for diabetes mellitus compared with individuals without HIV. Adipose tissue is an important regulator of glucose and lipid metabolism, and adipose tissue T cells modulate local inflammatory responses and, by ... ...

Abstract	Persons with HIV are at increased risk for diabetes mellitus compared with individuals without HIV. Adipose tissue is an important regulator of glucose and lipid metabolism, and adipose tissue T cells modulate local inflammatory responses and, by extension, adipocyte function. Persons with HIV and diabetes have a high proportion of CX3CR1
MeSH term(s)	Adipose Tissue/immunology ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cytomegalovirus/immunology ; Cytomegalovirus Infections/immunology ; Diabetes Mellitus/metabolism ; HIV Infections/immunology ; Humans ; Single-Cell Analysis/methods ; T-Lymphocyte Subsets/immunology
Language	English
Publishing date	2021-02-16
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2666-3791
ISSN (online)	2666-3791
DOI	10.1016/j.xcrm.2021.100205
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Article ; Online: Restricted myeloperoxidase epitopes drive the adaptive immune response in MPO-ANCA vasculitis.

Journal of autoimmunity

2019 Volume 106, Page(s) 102306

Abstract: Background: Treatment of autoimmune diseases has relied on broad immunosuppression. Knowledge of specific interactions between human leukocyte antigen (HLA), the autoantigen, and effector immune cells, provides the foundation for antigen-specific ... ...

Abstract	Background: Treatment of autoimmune diseases has relied on broad immunosuppression. Knowledge of specific interactions between human leukocyte antigen (HLA), the autoantigen, and effector immune cells, provides the foundation for antigen-specific therapies. These studies investigated the role of HLA, specific myeloperoxidase (MPO) epitopes, CD4 Methods: HLA sequence-based typing identified enriched alleles in our patient population (HLA-DPB104:01 and HLA-DRB401:01), while in silico and in vitro binding studies confirmed binding between HLA and specific MPO epitopes. Class II tetramers with MPO peptides were utilized to detect autoreactive CD4 Results: We identified a restricted region of MPO that was recognized by both CD4 Conclusions: These observations reveal interactions between restricted MPO epitopes and the adaptive immune system within ANCA vasculitis that may inform new antigen-specific therapies in autoimmune disease while providing insight into immunopathogenesis.
MeSH term(s)	Adaptive Immunity/immunology ; Amino Acid Sequence ; Animals ; Antibodies, Antineutrophil Cytoplasmic/immunology ; Autoantibodies/immunology ; Autoantigens/immunology ; CD4-Positive T-Lymphocytes/immunology ; Cells, Cultured ; Epitopes/immunology ; Humans ; Leukocytes, Mononuclear/immunology ; Longitudinal Studies ; Mice ; Peroxidase/immunology ; Receptors, Antigen, T-Cell/immunology ; Vasculitis/immunology
Chemical Substances	Antibodies, Antineutrophil Cytoplasmic ; Autoantibodies ; Autoantigens ; Epitopes ; Receptors, Antigen, T-Cell ; Peroxidase (EC 1.11.1.7)
Language	English
Publishing date	2019-08-02
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	639452-8
ISSN	1095-9157 ; 0896-8411
ISSN (online)	1095-9157
ISSN	0896-8411
DOI	10.1016/j.jaut.2019.102306
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