The human alloreactive CD4+ T-cell repertoire is biased to a Th17 response and the frequency is inversely related to the number of HLA class II mismatches

Abstract Estimates of precursor frequency and assessment of functional characteristics of alloreactive CD4+ T cells are all biased by the need for long-term culture. In this study, direct visualization of human alloreactive CD4+ T cells on the single-cell level was achieved using cell surface expression of CD154 as a tool for identification. The average frequency of alloreactive CD154+CD4+ T cells among peripheral blood CD4+ T cells was 0.1%, with half of the cells displaying a naive phenotype. The proliferation capacity and expression of cytokines after allogeneic stimulation resided in these CD154+CD4+ T cells. The repertoire of alloreactive CD4+ T cells was biased to a Th17 response, and on average 24% of alloreactive CD154+CD4+ memory T cells produced interleukin-17 (IL-17) after polyclonal stimulation. Unexpectedly, mixed cell cultures from human leukocyte antigen (HLA)–identical donors also generated alloreactive CD154+CD4+ T cells and yielded the highest frequency compared with HLA-nonidentical combinations. Therefore, reactivity to minor histocompatibility antigens between HLA-identical subjects appears to be relatively common. Alloreactive HLA-identical T cells did not proliferate or express cytokines, but were driven to proliferation in the presence of exogenous IL-2.

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Regulation of Cell Surface Expression of CTLA-4 by Secretion of CTLA-4-Containing Lysosomes Upon Activation of CD4+T Cells

The Journal of Immunology ◽

10.4049/jimmunol.165.9.5062 ◽

2000 ◽

Vol 165 (9) ◽

pp. 5062-5068 ◽

Cited By ~ 89

Author(s):

Tomohiko Iida ◽

Hiroshi Ohno ◽

Chiaki Nakaseko ◽

Machie Sakuma ◽

Mitsue Takeda-Ezaki ◽

...

Keyword(s):

T Cells ◽

Cell Surface ◽

Cd4 T Cells ◽

Surface Expression ◽

Cell Surface Expression

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Regulation of the cell-surface expression of an HTLV-I binding protein in human T cells during immune activation

Blood ◽

10.1182/blood-2002-07-2277 ◽

2003 ◽

Vol 101 (8) ◽

pp. 3085-3092 ◽

Cited By ~ 30

Author(s):

Manisha D. Nath ◽

Francis W. Ruscetti ◽

Cari Petrow-Sadowski ◽

Kathryn S. Jones

Keyword(s):

T Cells ◽

T Cell ◽

Cell Surface ◽

Cell Lines ◽

Cd4 T Cells ◽

Binding Protein ◽

Surface Expression ◽

Cell Surface Expression ◽

Type I ◽

Primary Cells

AbstractLittle is known about the requirements for human T-cell leukemia virus type I (HTLV-I) entry, including the identity of the cellular receptor(s). Recently, we have generated an HTLV-I surface glycoprotein (SU) immunoadhesin, HTSU-IgG, which binds specifically to cell-surface protein(s) critical for HTLV-I–mediated entry in cell lines. Here, expression of the HTLV-I SU binding protein on primary cells of the immune system was examined. The immunoadhesin specifically bound to adult T cells, B cells, NK cells, and macrophages. Cell stimulation dramatically increased the amount of binding, with the highest levels of binding on CD4+ and CD8+ T cells. Naive (CD45RAhigh, CD62Lhigh) CD4+ T cells derived from cord blood cells, in contrast to other primary cells and all cell lines examined, bound no detectable HTLV-I SU. However, following stimulation, the level of HTSU-IgG binding was rapidly induced (fewer than 6 hours), reaching the level of binding seen on adult CD4+ T cells by 72 hours. In contrast to HTLV-I virions, the soluble HTSU-IgG did not effect T-cell activation or proliferation. When incubated with human peripheral blood mononuclear cells in a mixed leukocyte reaction, HTSU-IgG inhibited proliferation at less than 1 ng/mL. These results indicate that cell-surface expression of the HTLV SU binding protein is up-regulated during in vitro activation and suggest a role for the HTLV-I SU binding proteins in the immunobiology of CD4+ T cells.

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Altered leukocyte response to CXCL12 in patients with warts hypogammaglobulinemia, infections, myelokathexis (WHIM) syndrome

Blood ◽

10.1182/blood-2003-10-3532 ◽

2004 ◽

Vol 104 (2) ◽

pp. 444-452 ◽

Cited By ~ 128

Author(s):

Anna Virginia Gulino ◽

Daniele Moratto ◽

Silvano Sozzani ◽

Patrizia Cavadini ◽

Karel Otero ◽

...

Keyword(s):

T Cells ◽

Chemokine Receptor ◽

Gene Mutations ◽

Surface Expression ◽

Cell Surface Expression ◽

Effector Memory ◽

Polymorphonuclear Cells ◽

Cell Repertoire ◽

Whim Syndrome ◽

Leukocyte Response

Abstract The chemokine receptor CXCR4 and its functional ligand, CXCL12, are essential regulators of development and homeostasis of hematopoietic and lymphoid organs. Heterozygous truncating mutations in the CXCR4 intracellular tail cause a rare genetic disease known as WHIM syndrome (warts, hypogammaglobulinemia, infections, myelokathexis), whose pathophysiology remains unclear. We report CXCR4 function in 3 patients with WHIM syndrome carrying heterozygous truncating mutations of CXCR4. We show that CXCR4 gene mutations in WHIM patients do not affect cell surface expression of the chemokine receptor and its internalization upon stimulation with CXCL12. Moreover, no significant differences in calcium mobilization in response to CXCL12 are found. However, the chemotactic response of both polymorphonuclear cells and T lymphocytes in response to CXCL12 is increased. Furthermore, immunophenotypic analysis of circulating T and B lymphocytes reveals a decreased number of memory B cells and of naive T cells and an accumulation of effector memory T cells associated with a restricted T-cell repertoire. Based on our results, we suggest that the altered leukocyte response to CXCL12 may account for the pathologic retention of mature polymorphonuclear cells in the bone marrow (myelokathexis) and for an altered lymphocyte trafficking, which may cause the immunophenotyping abnormalities observed in WHIM patients. (Blood. 2004;104:444-452)

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Transcriptional down-regulation of ccr5 in a subset of HIV+ controllers and their family members

eLife ◽

10.7554/elife.44360 ◽

2019 ◽

Vol 8 ◽

Cited By ~ 4

Author(s):

Elena Gonzalo-Gil ◽

Patrick B Rapuano ◽

Uchenna Ikediobi ◽

Rebecca Leibowitz ◽

Sameet Mehta ◽

...

Keyword(s):

T Cells ◽

T Cell Activation ◽

Cd4 T Cells ◽

Cell Activation ◽

Family Members ◽

Surface Expression ◽

Cell Surface Expression ◽

Down Regulation ◽

Control Virus ◽

Hiv Controllers

HIV +Elite and Viremic controllers (EC/VCs) are able to control virus infection, perhaps because of host genetic determinants. We identified 16% (21 of 131) EC/VCs with CD4 +T cells with resistance specific to R5-tropic HIV, reversed after introduction of ccr5. R5 resistance was not observed in macrophages and depended upon the method of T cell activation. CD4 +T cells of these EC/VCs had lower ccr2 and ccr5 RNA levels, reduced CCR2 and CCR5 cell-surface expression, and decreased levels of secreted chemokines. T cells had no changes in chemokine receptor mRNA half-life but instead had lower levels of active transcription of ccr2 and ccr5, despite having more accessible chromatin by ATAC-seq. Other nearby genes were also down-regulated, over a region of ~500 kb on chromosome 3p21. This same R5 resistance phenotype was observed in family members of an index VC, also associated with ccr2/ccr5 down-regulation, suggesting that the phenotype is heritable.

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Cyclooxygenase Regulates Cell Surface Expression of CXCR3/1-Storing Granules in Human CD4+T Cells

The Journal of Immunology ◽

10.4049/jimmunol.177.12.8806 ◽

2006 ◽

Vol 177 (12) ◽

pp. 8806-8812 ◽

Cited By ~ 9

Author(s):

Olivier Gasser ◽

Thomas A. Schmid ◽

Gabriela Zenhaeusern ◽

Christoph Hess

Keyword(s):

T Cells ◽

Cell Surface ◽

Cd4 T Cells ◽

Surface Expression ◽

Cell Surface Expression

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In vitro expansion of CMV-specific CD4+ T-cells by artificial antigen presentation for adoptive immunotherapy in the post- transplant setting

Journal of Clinical Oncology ◽

10.1200/jco.2007.25.18_suppl.3049 ◽

2007 ◽

Vol 25 (18_suppl) ◽

pp. 3049-3049

Author(s):

R. J. Ricafort ◽

M. Stephan ◽

R. O’Reilly ◽

M. Sadelain

Keyword(s):

T Cells ◽

Adoptive Immunotherapy ◽

Cd4 T Cells ◽

Flow Cytometric Analysis ◽

Surface Expression ◽

Cell Surface Expression ◽

Trypan Blue Exclusion ◽

Artificial Antigen ◽

Soluble Class

3049 Background: CMV continues to be a major cause of morbidity and mortality in marrow allograft recipients. The importance of CD4+ T-cells in maintaining immunity and therapeutic efficacy in adoptive cellular strategies has been appreciated. With a novel artificial antigen presenting cell (AAPC) system, we induced the expansion of CMV-specific CD4+ T-cells. Methods: AAPCs were generated by using a standardizable line of murine 3T3 cells sequentially transduced to express human ICAM-1, LFA-1, B7.1, HLA A*0201 and DRB1*1101 alleles. Antigen was provided either by peptide pulsing with the HLA-DRB1*1101-restricted, CMV- pp65-derived P92 peptide, or the influenza matrix peptide (IMP) as a control, or by transgene expression of pp65. PBMCs were isolated from a CMV+, HLA*A02, DRB1*11-positive healthy donor after informed consent, and enriched for T-cells via positive selection using sheep RBCs. Cocultures of T-cells with AAPCs were established at a ratio of 10:1. 20 IU/ml of IL-2 was added on d7 and every third day thereafter. On d12- 14 of co-culture, the T-cells were restimulated on a fresh monolayer of AAPCs with antigen. Cells were counted in triplicate via hemocytometer with trypan blue exclusion. Immunophenotyping of expanded T-cells was done via flow cytometric analysis for cell surface expression of CD3, CD8, CD4, and soluble Class II (HLA-DRB1*1101) tetramer specific for the P92 peptide of CMV-pp65. Results: Following 3 rounds of stimulation, antigen-specific fold expansion, defined by DR11,P92-tetramer-positive, CD4+ T-cells, was 313-fold when stimulated on AAPC.A2.DR11.PP65, compared to 5–35-fold using the other AAPC controls. Conclusion: We are able to expand clinically relevant numbers of CMV-specific CD4+ T-cells using a rapid, practicable, and broadly applicable approach. These preclinical studies show the feasibility of generating virus-specific T-cells of desired specificity and HLA restriction for adoptive immunotherapy of severe CMV infections. [Table: see text] No significant financial relationships to disclose.

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Influence of HLA-C Expression Level on HIV Control

Science ◽

10.1126/science.1232685 ◽

2013 ◽

Vol 340 (6128) ◽

pp. 87-91 ◽

Cited By ~ 247

Author(s):

Richard Apps ◽

Ying Qi ◽

Jonathan M. Carlson ◽

Haoyan Chen ◽

Xiaojiang Gao ◽

...

Keyword(s):

Cytotoxic T Lymphocyte ◽

Human Leukocyte ◽

Surface Expression ◽

Viral Escape ◽

Cell Surface Expression ◽

Escape Mutation ◽

Expression Levels ◽

European Americans ◽

Genome Wide ◽

Lymphocyte Responses

A variant upstream of human leukocyte antigen C (HLA-C) shows the most significant genome-wide effect on HIV control in European Americans and is also associated with the level of HLA-C expression. We characterized the differential cell surface expression levels of all common HLA-C allotypes and tested directly for effects of HLA-C expression on outcomes of HIV infection in 5243 individuals. Increasing HLA-C expression was associated with protection against multiple outcomes independently of individual HLA allelic effects in both African and European Americans, regardless of their distinct HLA-C frequencies and linkage relationships with HLA-B and HLA-A. Higher HLA-C expression was correlated with increased likelihood of cytotoxic T lymphocyte responses and frequency of viral escape mutation. In contrast, high HLA-C expression had a deleterious effect in Crohn’s disease, suggesting a broader influence of HLA expression levels in human disease.

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T cell receptor recognition of hybrid insulin peptides bound to HLA-DQ8

Nature Communications ◽

10.1038/s41467-021-25404-x ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Mai T. Tran ◽

Pouya Faridi ◽

Jia Jia Lim ◽

Yi Tian Ting ◽

Goodluck Onwukwe ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cd4 T Cells ◽

Type I Diabetes ◽

Binding Motif ◽

Type I ◽

Cell Repertoire ◽

Islet Amyloid ◽

Autoreactive T Cell ◽

Β Chain

AbstractHLA-DQ8, a genetic risk factor in type I diabetes (T1D), presents hybrid insulin peptides (HIPs) to autoreactive CD4+ T cells. The abundance of spliced peptides binding to HLA-DQ8 and how they are subsequently recognised by the autoreactive T cell repertoire is unknown. Here we report, the HIP (GQVELGGGNAVEVLK), derived from splicing of insulin and islet amyloid polypeptides, generates a preferred peptide-binding motif for HLA-DQ8. HLA-DQ8-HIP tetramer+ T cells from the peripheral blood of a T1D patient are characterised by repeated TRBV5 usage, which matches the TCR bias of CD4+ T cells reactive to the HIP peptide isolated from the pancreatic islets of a patient with T1D. The crystal structure of three TRBV5+ TCR-HLA-DQ8-HIP complexes shows that the TRBV5-encoded TCR β-chain forms a common landing pad on the HLA-DQ8 molecule. The N- and C-termini of the HIP is recognised predominantly by the TCR α-chain and TCR β-chain, respectively, in all three TCR ternary complexes. Accordingly, TRBV5 + TCR recognition of HIP peptides might occur via a ‘polarised’ mechanism, whereby each chain within the αβTCR heterodimer recognises distinct origins of the spliced peptide presented by HLA-DQ8.

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