Reduced polyfunctional T cells and increased cellular activation markers in adult allergy patients reporting adverse reactions to food

Abstract Background: The underlying cellular mechanisms causing adverse reactions to food are complex and still not fully understood. Therefore, in this study we aimed to identify functional and/or phenotypical immune cell signatures characteristic for adult patients reporting adverse reactions to food. By mass cytometry, we performed high-dimensional profiling of peripheral blood mononuclear cells (PBMC) from adult patients reporting adverse reactions to food and healthy controls. The patients were grouped according to sIgE-positive or sIgE-negative serology to common food and inhalant allergens. Two broad antibody panels were used, allowing determination of major immune cell populations in PBMC, as well as activation status, proliferation status, and cytokine expression patterns after PMA/ionomycin-stimulation on a single cell level. Results: By use of data-driven algorithms, several cell populations were identified showing significantly different marker expression between the groups. Most striking was an impaired frequency and function of polyfunctional CD4 + and CD8 + T cells in patients reporting adverse reactions to food compared to the controls. Further, subpopulations of monocytes, T cells, and B cells had increased expression of functional markers such as CD371, CD69, CD25, CD28, and/or HLA-DR as well as decreased expression of CD23 in the patients. Most of the differing cell subpopulations were similarly altered in the two subgroups of patients. Conclusion: Our results suggest common immune cell features for both patient subgroups reporting adverse reactions to food, and provide a basis for further studies on mechanistic and diagnostic biomarker studies in food allergy.

Download Full-text

Reduced polyfunctional T cells and increased cellular activation markers in adult allergy patients reporting adverse reactions to food

10.21203/rs.2.23606/v2 ◽

2020 ◽

Author(s):

Friederike Sonnet ◽

Ellen Namork ◽

Eva Stylianou ◽

Ingvild Gaare-Olstad ◽

Kanutte Huse ◽

...

Keyword(s):

T Cells ◽

Adverse Reactions ◽

Mononuclear Cells ◽

Immune Cell ◽

Expression Patterns ◽

Cell Populations ◽

Functional Markers ◽

Cellular Mechanisms ◽

Peripheral Blood Mononuclear ◽

Activation Markers

Abstract Background: The underlying cellular mechanisms causing adverse reactions to food are complex and still not fullyunderstood. Therefore, in this study we aimed to identify functional and/or phenotypical immune cell signaturescharacteristic for adult patients reporting adverse reactions to food.By mass cytometry, we performed high-dimensional profiling of peripheral blood mononuclear cells (PBMC) fromadult patients reporting adverse reactions to food and healthy controls. The patients were grouped according to sIgE-positive or sIgE-negative serology to common food and inhalant allergens. Two broad antibody panels were used,allowing determination of major immune cell populations in PBMC, as well as activation status, proliferation status,and cytokine expression patterns after PMA/ionomycin-stimulation on a single cell level.Results: By use of data-driven algorithms, several cell populations were identified showing significantly differentmarker expression between the groups.Most striking was an impaired frequency and function of polyfunctional CD4+ and CD8+ T cells in patients reportingadverse reactions to food compared to the controls. Further, subpopulations of monocytes, T cells, and B cells hadincreased expression of functional markers such as CD371, CD69, CD25, CD28, and/or HLA-DR as well asdecreased expression of CD23 in the patients. Most of the differing cell subpopulations were similarly altered in thetwo subgroups of patients.Conclusion: Our results suggest common immune cell features for both patient subgroups reporting adversereactions to food, and provide a basis for further studies on mechanistic and diagnostic biomarker studies in foodallergy.

Download Full-text

Reduced polyfunctional T cells and increased cellular activation markers in adult allergy patients reporting adverse reactions to food

10.21203/rs.2.23606/v3 ◽

2020 ◽

Author(s):

Friederike Sonnet ◽

Ellen Namork ◽

Eva Stylianou ◽

Ingvild Gaare-Olstad ◽

Kanutte Huse ◽

...

Keyword(s):

T Cells ◽

Adverse Reactions ◽

Mononuclear Cells ◽

Immune Cell ◽

Expression Patterns ◽

Cell Populations ◽

Functional Markers ◽

Cellular Mechanisms ◽

Peripheral Blood Mononuclear ◽

Activation Markers

Abstract Background: The underlying cellular mechanisms causing adverse reactions to food are complex and still not fullyunderstood. Therefore, in this study we aimed to identify functional and/or phenotypical immune cell signaturescharacteristic for adult patients reporting adverse reactions to food.By mass cytometry, we performed high-dimensional profiling of peripheral blood mononuclear cells (PBMC) fromadult patients reporting adverse reactions to food and healthy controls. The patients were grouped according to sIgE-positive or sIgE-negative serology to common food and inhalant allergens. Two broad antibody panels were used,allowing determination of major immune cell populations in PBMC, as well as activation status, proliferation status,and cytokine expression patterns after PMA/ionomycin-stimulation on a single cell level.Results: By use of data-driven algorithms, several cell populations were identified showing significantly differentmarker expression between the groups.Most striking was an impaired frequency and function of polyfunctional CD4+ and CD8+ T cells in patients reportingadverse reactions to food compared to the controls. Further, subpopulations of monocytes, T cells, and B cells hadincreased expression of functional markers such as CD371, CD69, CD25, CD28, and/or HLA-DR as well asdecreased expression of CD23 in the patients. Most of the differing cell subpopulations were similarly altered in thetwo subgroups of patients.Conclusion: Our results suggest common immune cell features for both patient subgroups reporting adversereactions to food, and provide a basis for further studies on mechanistic and diagnostic biomarker studies in foodallergy.

Download Full-text

Faculty Opinions recommendation of Polyfunctional T cells accumulate in large human cytomegalovirus-specific T cell responses.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13371073.14741210 ◽

2011 ◽

Author(s):

Holden Maecker

Keyword(s):

T Cells ◽

T Cell ◽

Human Cytomegalovirus ◽

T Cell Responses ◽

Cell Responses ◽

Polyfunctional T Cells

Download Full-text

12 Development of an in vitro assay to assess bispecific T cell engager using T cells from CD3e humanized mice

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0012 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A12-A12

Author(s):

Jun Zhou ◽

Shuang Zhu ◽

Hongjuan Zhang ◽

Lei Zheng ◽

Mingfa Zang ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Tumor Cells ◽

Lymphoblastic Leukemia ◽

Humanized Mice ◽

Activation Markers ◽

In Vivo Models ◽

Vitro Assay ◽

Bite Antibody

BackgroundBispecific T cell engagers (BiTE) is a fast-growing class of immunotherapies. They are bispecific antibody that bind to T cell-surface protein (for example, CD3e) and a specific tumor associate antigen (TAA) on tumor cells, by which to redirect T cells against tumor cells in a MHC-independent manner. A successful example in the clinical is Blinatumomab, a BiTE antibody against CD3/CD19 approved in 2014 to treat acute lymphoblastic leukemia. Currently, many CD3-based BiTE are in clinical trials, including BCMAxCD3, Her2xCD3, CEAxCD3, and PSMAxCD3. To evaluate the efficacy of BiTE in vitro, human peripheral blood monocyte cells (hPBMC) are commonly being used as a source of T cells to co-culture with tumor cells. The disadvantage of using hPBMC is donor-to-donor variability and the availability of the original donor if a study needs to be repeated.MethodsTo overcome this, we proposed to replace hPBMC with T cells from human CD3e (hCD3) genetically engineered mouse models mice (GEMM) for in in vitro coculture assay. T cells were isolated from hCD3 GEMM mice using negative selection mouse T cell isolation kit. Conventional tumor cell lines or luciferase-engineered patient-derived-xenograft (PDX)-derived organoids (PDXO) expressing specific antigens are co-cultured with hCD3 T cells in 96-well plates in the presence of BiTE antibody.ResultsWe measured the killing of tumor cells using either flow cytometry or luciferase activity as readouts. To analyze tumor-reactivity of T cells to cancer cell line or organoids, IFN-gamma in the culture medium was measured and activation markers on T cells was assessed.ConclusionsOur data showed the feasibility of using humanized mice T cells as a replacement for hPBMCs to assess BiTE antibody in vitro. We are further validating the application of murine hCD3 T cells for in vivo models to test bispecific T cell engagers.

Download Full-text

CCL21 mediates CD4+ T-cell costimulation via a DOCK2/Rac-dependent pathway

Blood ◽

10.1182/blood-2009-01-200923 ◽

2009 ◽

Vol 114 (3) ◽

pp. 580-588 ◽

Cited By ~ 45

Author(s):

Kathrin Gollmer ◽

François Asperti-Boursin ◽

Yoshihiko Tanaka ◽

Klaus Okkenhaug ◽

Bart Vanhaesebroeck ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Activation ◽

Cd4 T Cells ◽

Cell Activation ◽

Early Time ◽

Cell Receptor ◽

Tcr Signaling ◽

Activation Markers

Abstract CD4+ T cells use the chemokine receptor CCR7 to home to and migrate within lymphoid tissue, where T-cell activation takes place. Using primary T-cell receptor (TCR)–transgenic (tg) CD4+ T cells, we explored the effect of CCR7 ligands, in particular CCL21, on T-cell activation. We found that the presence of CCL21 during early time points strongly increased in vitro T-cell proliferation after TCR stimulation, correlating with increased expression of early activation markers. CCL21 costimulation resulted in increased Ras- and Rac-GTP formation and enhanced phosphorylation of Akt, MEK, and ERK but not p38 or JNK. Kinase-dead PI3KδD910A/D910A or PI3Kγ-deficient TCR-tg CD4+ T cells showed similar responsiveness to CCL21 costimulation as control CD4+ T cells. Conversely, deficiency in the Rac guanine exchange factor DOCK2 significantly impaired CCL21-mediated costimulation in TCR-tg CD4+ T cells, concomitant with impaired Rac- but not Ras-GTP formation. Using lymph node slices for live monitoring of T-cell behavior and activation, we found that G protein-coupled receptor signaling was required for early CD69 expression but not for Ca2+ signaling. Our data suggest that the presence of CCL21 during early TCR signaling lowers the activation threshold through Ras- and Rac-dependent pathways leading to increased ERK phosphorylation.

Download Full-text

Generation of primary antigen-specific human cytotoxic T lymphocytes in human/mouse radiation chimera

Blood ◽

10.1182/blood.v88.2.721.bloodjournal882721 ◽

1996 ◽

Vol 88 (2) ◽

pp. 721-730 ◽

Cited By ~ 30

Author(s):

H Segall ◽

I Lubin ◽

H Marcus ◽

A Canaan ◽

Y Reisner

Keyword(s):

T Cells ◽

T Cell ◽

Human Lymphocytes ◽

Scid Mice ◽

Human Antibody ◽

Adoptive Therapy ◽

Activation Markers ◽

Human T Cell ◽

Human T Cells

Severe combined immunodeficient (SCID) mice are increasingly used as hosts for the adoptive transfer of human lymphocytes. Human antibody responses can be obtained in these xenogeneic chimeras, but information about the functionality of the human T cells in SCID mice is limited and controversial. Studies using human peripheral blood lymphocytes (PBL) injected intraperitoneally (IP) into SCID mice (hu-PBL-SCID mice) have shown that human T cells from these chimeras are anergic and have a defective signaling via the T-cell receptor. In addition, their antigenic repertoire is limited to xenoreactive clones. In the present study, we tested the functionality of human T cell in a recently described chimeric model. In this system, BALB/c mice are conditioned by irradiation and then transplanted with SCID bone marrow, followed by IP injection of human PBL. Our experiments demonstrated that human T cells, recovered from these hu-PBL-BALB mice within 1 month posttransplant, proliferated and expressed activation markers upon stimulation with anti-CD3 monoclonal antibody. A vigorous antiallogeneic human cytotoxic T-lymphocyte (CTL) response could be generated in these mice by immunizing them with irradiated allogeneic cells. Moreover, anti-human immunodeficiency virus type 1 (HIV-1) Net- specific human CTLs could be generated in vivo from naive lymphocytes by immunization of mouse-human chimeras with a recombinant vaccinia-nef virus. This model may be used to evaluate potential immunomodulatory drugs or cytokines, and could provide a relevant model for testing HIV vaccines, for production of antiviral T-cell clones for adoptive therapy, and for studying human T-cell responses in vivo.

Download Full-text

CD8+ T Cells Mediate Recovery andImmunopathology in West Nile VirusEncephalitis

Journal of Virology ◽

10.1128/jvi.77.24.13323-13334.2003 ◽

2003 ◽

Vol 77 (24) ◽

pp. 13323-13334 ◽

Cited By ~ 226

Author(s):

Yang Wang ◽

Mario Lobigs ◽

Eva Lee ◽

Arno Müllbacher

Keyword(s):

T Cells ◽

Dose Group ◽

Low Dose ◽

Neuronal Degeneration ◽

High Dose ◽

West Nile ◽

Activation Markers ◽

High Doses ◽

The Brain ◽

Deficient Mice

ABSTRACT C57BL/6J mice infected intravenously with the Sarafend strain of West Nile virus (WNV) develop a characteristic central nervous system (CNS) disease, including an acute inflammatory reaction. Dose response studies indicate two distinct kinetics of mortality. At high doses of infection (108 PFU), direct infection of the brain occurred within 24 h, resulting in 100% mortality with a 6-day mean survival time (MST), and there was minimal destruction of neural tissue. A low dose (103 PFU) of infection resulted in 27% mortality (MST, 11 days), and virus could be detected in the CNS 7 days postinfection (p.i.). Virus was present in the hypogastric lymph nodes and spleens at days 4 to 7 p.i. Histology of the brains revealed neuronal degeneration and inflammation within leptomeninges and brain parenchyma. Inflammatory cell infiltration was detectable in brains from day 4 p.i. onward in the high-dose group and from day 7 p.i. in the low-dose group, with the severity of infiltration increasing over time. The cellular infiltrates in brain consisted predominantly of CD8+, but not CD4+, T cells. CD8+ T cells in the brain and the spleen expressed the activation markers CD69 early and expressed CD25 at later time points. CD8+ T-cell-deficient mice infected with 103 PFU of WNV showed increased mortalities but prolonged MST and early infection of the CNS compared to wild-type mice. Using high doses of virus in CD8-deficient mice leads to increased survival. These results provide evidence that CD8+ T cells are involved in both recovery and immunopathology in WNV infection.

Download Full-text

A phase I vaccination study with dendritic cells loaded with NY-ESO-1 and α-galactosylceramide: induction of polyfunctional T cells in high-risk melanoma patients

Cancer Immunology Immunotherapy ◽

10.1007/s00262-017-2085-9 ◽

2017 ◽

Vol 67 (2) ◽

pp. 285-298 ◽

Cited By ~ 22

Author(s):

Olivier Gasser ◽

Katrina J. Sharples ◽

Catherine Barrow ◽

Geoffrey M. Williams ◽

Evelyn Bauer ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

High Risk ◽

Phase I ◽

High Risk Melanoma ◽

Risk Melanoma ◽

Melanoma Patients ◽

Polyfunctional T Cells

Download Full-text

TRPM2 Is Not Required for T-Cell Activation and Differentiation

Frontiers in Immunology ◽

10.3389/fimmu.2021.778916 ◽

2022 ◽

Vol 12 ◽

Author(s):

Niels C. Lory ◽

Mikolaj Nawrocki ◽

Martina Corazza ◽

Joanna Schmid ◽

Valéa Schumacher ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Activation ◽

Cell Activation ◽

Transient Receptor Potential ◽

Cell Function ◽

Intestinal Inflammation ◽

Activation Markers

Antigen recognition by the T-cell receptor induces a cytosolic Ca2+ signal that is crucial for T-cell function. The Ca2+ channel TRPM2 (transient receptor potential cation channel subfamily M member 2) has been shown to facilitate influx of extracellular Ca2+ through the plasma membrane of T cells. Therefore, it was suggested that TRPM2 is involved in T-cell activation and differentiation. However, these results are largely derived from in vitro studies using T-cell lines and non-physiologic means of TRPM2 activation. Thus, the relevance of TRPM2-mediated Ca2+ signaling in T cells remains unclear. Here, we use TRPM2-deficient mice to investigate the function of TRPM2 in T-cell activation and differentiation. In response to TCR stimulation in vitro, Trpm2-/- and WT CD4+ and CD8+ T cells similarly upregulated the early activation markers NUR77, IRF4, and CD69. We also observed regular proliferation of Trpm2-/- CD8+ T cells and unimpaired differentiation of CD4+ T cells into Th1, Th17, and Treg cells under specific polarizing conditions. In vivo, Trpm2-/- and WT CD8+ T cells showed equal specific responses to Listeria monocytogenes after infection of WT and Trpm2-/- mice and after transfer of WT and Trpm2-/- CD8+ T cells into infected recipients. CD4+ T-cell responses were investigated in the model of anti-CD3 mAb-induced intestinal inflammation, which allows analysis of Th1, Th17, Treg, and Tr1-cell differentiation. Here again, we detected similar responses of WT and Trpm2-/- CD4+ T cells. In conclusion, our results argue against a major function of TRPM2 in T-cell activation and differentiation.

Download Full-text