Large-Scale Prospective T Cell Function Assays in Shipped, Unfrozen Blood Samples: Experiences from the Multicenter TRIGR Trial

ABSTRACTBroad consensus assigns T lymphocytes fundamental roles in inflammatory, infectious, and autoimmune diseases. However, clinical investigations have lacked fully characterized and validated procedures, equivalent to those of widely practiced biochemical tests with established clinical roles, for measuring core T cell functions. The Trial to Reduce Insulin-dependent diabetes mellitus in the Genetically at Risk (TRIGR) type 1 diabetes prevention trial used consecutive measurements of T cell proliferative responses in prospectively collected fresh heparinized blood samples shipped by courier within North America. In this article, we report on the quality control implications of this simple and pragmatic shipping practice and the interpretation of positive- and negative-control analytes in our assay. We used polyclonal and postvaccination responses in 4,919 samples to analyze the development of T cell immunocompetence. We have found that the vast majority of the samples were viable up to 3 days from the blood draw, yet meaningful responses were found in a proportion of those with longer travel times. Furthermore, the shipping time of uncooled samples significantly decreased both the viabilities of the samples and the unstimulated cell counts in the viable samples. Also, subject age was significantly associated with the number of unstimulated cells and T cell proliferation to positive activators. Finally, we observed a pattern of statistically significant increases in T cell responses to tetanus toxin around the timing of infant vaccinations. This assay platform and shipping protocol satisfy the criteria for robust and reproducible long-term measurements of human T cell function, comparable to those of established blood biochemical tests. We present a stable technology for prospective disease-relevant T cell analysis in immunological diseases, vaccination medicine, and measurement of herd immunity.

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Deltex1 suppresses T cell function and is a biomarker for diagnosis and disease activity of systemic lupus erythematosus

Rheumatology ◽

10.1093/rheumatology/key418 ◽

2019 ◽

Vol 58 (4) ◽

pp. 719-728 ◽

Cited By ~ 1

Author(s):

Chuen-Miin Leu ◽

Tzu-Sheng Hsu ◽

Yu-Ping Kuo ◽

Ming-Zong Lai ◽

Po-Chun Liu ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Disease Activity ◽

Cell Function ◽

Healthy Controls ◽

Cell Functions ◽

T Cell Function ◽

Human T Cell ◽

Human T Cells ◽

Ifn Γ

Abstract Objective Deletion of Deltex1 (DTX1) in mice caused hyperactivation of T cells and lupus-like autoimmune syndromes, however, the association of DTX1 with human autoimmune diseases is totally unknown. This study investigated the role of DTX1 in human T cell functions and its correlation with disease activity in patients with SLE. Methods The influence of DTX1 on T cell function was evaluated using human primary cells. DTX1 expression in peripheral blood mononuclear cells (PBMCs) from healthy controls and SLE patients was measured by quantitative real-time PCR and the SLEDAI was used to assess disease activity. Results After stimulation with anti-CD3 and anti-CD28, silencing of DTX1 expression enhanced IFN-γ secretion by human T cells. The expression of DTX1 in PBMCs was significantly lower in 100 SLE patients than in 50 age- and sex-matched healthy controls (DTX1/glyceraldehyde 3-phosphate dehydrogenase, 0.452 vs 1.269, P < 0.001). The area under the receiver operator characteristics curve of the model was 0.737 (95% CI 0.658, 0.815). Intriguingly, a low DTX1 level in T cells led to high IFN-γ production in SLE patients and had a correlation with severe disease activity. In addition, low DTX1 expression in SLE patients was associated with active LN, lung involvement or hypocomplementaemia. Conclusion Knockdown DTX1 expression in human T cells reduced IFN-γ secretion. DTX1 expression in the PBMCs was significantly lower in SLE patients and had an inverse correlation with disease activity, indicating that the DTX1 level may be a good disease marker of SLE.

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Comparative effect of two pan-class I PI3K inhibitors used as anticancer drugs on human T cell function

International Immunopharmacology ◽

10.1016/j.intimp.2015.07.032 ◽

2015 ◽

Vol 28 (1) ◽

pp. 675-685 ◽

Cited By ~ 8

Author(s):

Belén Blanco ◽

Carmen Herrero-Sánchez ◽

Concepción Rodríguez-Serrano ◽

Mercedes Sánchez-Barba ◽

María Consuelo del Cañizo

Keyword(s):

T Cell ◽

Anticancer Drugs ◽

Cell Function ◽

Class I ◽

Comparative Effect ◽

Pi3k Inhibitors ◽

T Cell Function ◽

Human T Cell

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Allelic Variation of MHC Structure Alters Peptide Ligands to Induce Atypical Partial Agonistic CD8+ T Cell Function

Journal of Experimental Medicine ◽

10.1084/jem.20021796 ◽

2003 ◽

Vol 198 (1) ◽

pp. 99-109 ◽

Cited By ~ 10

Author(s):

Dong-Gyun Lim ◽

Jacqueline M. Slavik ◽

Katarzyna Bourcier ◽

Kathrine J. Smith ◽

David A. Hafler

Keyword(s):

T Cell ◽

Intracellular Signaling ◽

Cell Function ◽

Peptide Ligands ◽

T Cell Clones ◽

Altered Peptide Ligands ◽

Mhc Molecules ◽

Cell Functions ◽

T Cell Function ◽

Cell Clones

T cell receptors recognize small changes in peptide ligands leading to different T cell responses. Here, we analyzed a panel of HLA-A2–Tax11-19 reactive T cell clones to examine how small allelic variations of MHC molecules could alter the functional outcome of antigen recognition. Similar to the effects induced by antigenic altered peptide ligands, weak or partial agonistic T cell functions were identified in individual T cell clones with the recognition of MHC-altered peptide ligands (MAPLs). Interestingly, one subtype of HLA-A2 molecules induced an unusual type of partial agonistic function; proliferation without cytotoxicity. Modeling of crystallographic data indicated that polymorphic amino acids in the HLA-A2 peptide binding groove, especially the D-pocket, were responsible for this partial agonism. Reciprocal mutations of the Tax peptide side chain engaging the D-pocket indeed restored the agonist functions of the MHC–peptide complex. Whereas early intracellular signaling events were not efficiently induced by these MAPLs, phosphorylated c-Jun slowly accumulated with sustained long-term expression. These data indicate that MAPLs can induce atypical partial agonistic T cell function through structural and biochemical mechanisms similar to altered peptide ligands.

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HIV Patients Developing Primary CNS Lymphoma Lack EBV-Specific CD4+ T Cell Function Irrespective of Absolute CD4+ T Cell Counts

PLoS Medicine ◽

10.1371/journal.pmed.0040096 ◽

2007 ◽

Vol 4 (3) ◽

pp. e96 ◽

Cited By ~ 56

Author(s):

Olivier Gasser ◽

Florian K Bihl ◽

Marcel Wolbers ◽

Elisabetta Loggi ◽

Ingrid Steffen ◽

...

Keyword(s):

T Cell ◽

Cell Function ◽

Primary Cns Lymphoma ◽

Cd4 T Cell ◽

Cns Lymphoma ◽

Hiv Patients ◽

Cell Counts ◽

T Cell Function

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Paradoxical inhibition of T-cell function in response to CTLA-4 blockade; heterogeneity within the human T-cell population

Nature Medicine ◽

10.1038/72323 ◽

2000 ◽

Vol 6 (2) ◽

pp. 211-214 ◽

Cited By ~ 50

Author(s):

David E. Anderson ◽

Katarzyna D. Bieganowska ◽

Amit Bar-Or ◽

Enedina M.L. Oliveira ◽

Beatriz Carreno ◽

...

Keyword(s):

T Cell ◽

Cell Population ◽

Cell Function ◽

T Cell Function ◽

Human T Cell

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ALTERATION OF HUMAN T-CELL FUNCTION AFTER THYMIC IRRADIATION

Pediatric Research ◽

10.1203/00006450-197404000-00467 ◽

1974 ◽

Vol 8 (4) ◽

pp. 418-418

Author(s):

Christian H L Rieger ◽

Simmer C Kraft ◽

Richard M Rothberg

Keyword(s):

T Cell ◽

Cell Function ◽

T Cell Function ◽

Human T Cell

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Sprouty2 positively regulates T cell function and airway inflammation through regulation of CSK and LCK kinases

PLoS Biology ◽

10.1371/journal.pbio.3001063 ◽

2021 ◽

Vol 19 (3) ◽

pp. e3001063

Author(s):

Anand Sripada ◽

Kapil Sirohi ◽

Lidia Michalec ◽

Lei Guo ◽

Jerome T. McKay ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cell Function ◽

Airway Hyperreactivity ◽

Mouse Strains ◽

Caveolin 1 ◽

T Cell Function ◽

Human T Cell ◽

Extracellular Signal

The function of Sprouty2 (Spry2) in T cells is unknown. Using 2 different (inducible and T cell–targeted) knockout mouse strains, we found that Spry2 positively regulated extracellular signal-regulated kinase 1/2 (ERK1/2) signaling by modulating the activity of LCK. Spry2−/− CD4+ T cells were unable to activate LCK, proliferate, differentiate into T helper cells, or produce cytokines. Spry2 deficiency abrogated type 2 inflammation and airway hyperreactivity in a murine model of asthma. Spry2 expression was higher in blood and airway CD4+ T cells from patients with asthma, and Spry2 knockdown impaired human T cell proliferation and cytokine production. Spry2 deficiency up-regulated the lipid raft protein caveolin-1, enhanced its interaction with CSK, and increased CSK interaction with LCK, culminating in augmented inhibitory phosphorylation of LCK. Knockdown of CSK or dislodgment of caveolin-1–bound CSK restored ERK1/2 activation in Spry2−/− T cells, suggesting an essential role for Spry2 in LCK activation and T cell function.

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Human T-cell function in experimental ascorbic acid deficiency and spontaneous scurvy

American Journal of Clinical Nutrition ◽

10.1093/ajcn/36.1.127 ◽

1982 ◽

Vol 36 (1) ◽

pp. 127-130 ◽

Cited By ~ 14

Author(s):

N E Kay ◽

D E Holloway ◽

S W Hutton ◽

N D Bone ◽

W C Duane

Keyword(s):

Ascorbic Acid ◽

T Cell ◽

Cell Function ◽

Ascorbic Acid Deficiency ◽

T Cell Function ◽

Human T Cell ◽

Acid Deficiency

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Recombinant CD95-Fc (APG101) prevents graft-versus-host disease in mice without disabling antitumor cytotoxicity and T-cell functions

Blood ◽

10.1182/blood-2012-04-423392 ◽

2013 ◽

Vol 121 (3) ◽

pp. 556-565 ◽

Cited By ~ 6

Author(s):

Natalie Hartmann ◽

Joanna J. Messmann ◽

Frank Leithäuser ◽

Maxi Weiswange ◽

Michael Kluge ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Graft Versus Host Disease ◽

Cell Function ◽

Allogeneic Bone Marrow Transplantation ◽

Allogeneic Bone ◽

Host Disease ◽

Graft Versus Host ◽

Cell Functions ◽

T Cell Function

Abstract Graft-versus-host disease (GVHD) induced by transplant-derived T cells represents a major complication after allogeneic bone marrow transplantation (BMT). However, these T cells support engraftment, early T-cell immunity, and mediate the graft-versus-tumor (GVT) effect. Cytotoxic effector functions by transplanted T cells are predominantly mediated by the perforin/granzyme and the CD95/CD95L system. APG101, a novel recombinant human fusion protein consisting of the extracellular domain of CD95 and the Fc domain of an IgG1 antibody inhibited CD95L-induced apoptosis without interfering with T-cell function in vitro and was therefore tested for its ability to prevent GVHD in murine BMT models across minor or major histocompatibility barriers. Starting APG101 treatment either 1 day before or 6 days after transplantation effectively reduced clinical GVHD and rescued survival between 60% and 100% if GVHD was CD95L mediated. APG101 did not interfere with the GVT effect, because P815 mastocytoma and most importantly primary Bcr-Abl–transformed B-cell leukemias were completely eradicated by the alloantigen-specific T cells. Phenotype and homing of alloantigen-specific T cells or their perforin/granzyme-mediated cytotoxicity and proliferative capacity were not affected by APG101 treatment suggesting that APG101 therapy might be useful in GVHD prophylaxis without impairing T-cell function and most importantly preserving GVT activity.

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