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

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)

Download Full-text

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

Blood ◽

10.1182/blood-2009-03-211896 ◽

2009 ◽

Vol 114 (18) ◽

pp. 3947-3955 ◽

Cited By ~ 17

Author(s):

Nicolle H. R. Litjens ◽

Jacqueline van de Wetering ◽

Nicole M. van Besouw ◽

Michiel G. H. Betjes

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Human Leukocyte ◽

Surface Expression ◽

Average Frequency ◽

Cell Surface Expression ◽

Interleukin 17 ◽

Cell Repertoire ◽

Mixed Cell ◽

Th17 Response

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.

Download Full-text

Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells

Journal of Experimental Medicine ◽

10.1084/jem.20060468 ◽

2006 ◽

Vol 203 (7) ◽

pp. 1693-1700 ◽

Cited By ~ 1056

Author(s):

Nabila Seddiki ◽

Brigitte Santner-Nanan ◽

Jeff Martinson ◽

John Zaunders ◽

Sarah Sasson ◽

...

Keyword(s):

T Cells ◽

Peripheral Blood ◽

Surface Expression ◽

Cell Surface Expression ◽

Effector Memory ◽

Accurate Estimation ◽

Activated T Cells ◽

Interleukin 7 ◽

Highly Correlated

Abnormalities in CD4+CD25+Foxp3+ regulatory T (T reg) cells have been implicated in susceptibility to allergic, autoimmune, and immunoinflammatory conditions. However, phenotypic and functional assessment of human T reg cells has been hampered by difficulty in distinguishing between CD25-expressing activated and regulatory T cells. Here, we show that expression of CD127, the α chain of the interleukin-7 receptor, allows an unambiguous flow cytometry–based distinction to be made between CD127lo T reg cells and CD127hi conventional T cells within the CD25+CD45RO+RA− effector/memory and CD45RA+RO− naive compartments in peripheral blood and lymph node. In healthy volunteers, peripheral blood CD25+CD127lo cells comprised 6.35 ± 0.26% of CD4+ T cells, of which 2.05 ± 0.14% expressed the naive subset marker CD45RA. Expression of FoxP3 protein and the CD127lo phenotype were highly correlated within the CD4+CD25+ population. Moreover, both effector/memory and naive CD25+CD127lo cells manifested suppressive activity in vitro, whereas CD25+CD127hi cells did not. Cell surface expression of CD127 therefore allows accurate estimation of T reg cell numbers and isolation of pure populations for in vitro studies and should contribute to our understanding of regulatory abnormalities in immunopathic diseases.

Download Full-text

Cattle T Cell Phenotyping by an 8-Colour, 10-Parameter Panel

10.1101/2022.01.12.476027 ◽

2022 ◽

Author(s):

Eduard Otto Roos ◽

William Mwangi ◽

Wilhelm Gerner ◽

Ryan Waters ◽

John A Hammond

Keyword(s):

T Cells ◽

T Cell ◽

Mononuclear Cells ◽

Surface Expression ◽

T Cell Response ◽

Cell Surface Expression ◽

Effector Memory ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells ◽

Terminal Effector

This multiplex staining panel was developed to differentiate cattle T cells into conventional (CD4 and CD8) and unconventional (γδ-TCR) subsets as well as their stage of differentiation and activation. The combination of CD45RO and CD62L allows the identification of naïve (TNaïve), central memory (TCM), effector memory (TEM) and terminal effector (TTE) T cells. Activated cattle T cells (TAV) can be identified by the cell surface expression of CD25. This panel was developed using cryopreserved cattle peripheral blood mononuclear cells (PBMCs) and tested on fresh as well as stimulated PBMCs. Therefore, this 8-colour, 10-parameter flow cytometry panel simultaneously identifies cattle TNaïve, TAV, TCM, TEM, TTE and γδ-TCR cells. This panel will improve our ability to examine T cell response to pathogens and vaccines in cattle including the potential to identify previously undescribed subpopulations. Furthermore, this panel can be readily optimised for other bovid species as many of these reagents are likely to cross react.

Download Full-text

IL-8 responsiveness defines a subset of CD8 T cells poised to kill

Blood ◽

10.1182/blood-2004-03-1067 ◽

2004 ◽

Vol 104 (12) ◽

pp. 3463-3471 ◽

Cited By ~ 56

Author(s):

Christoph Hess ◽

Terry K. Means ◽

Patrick Autissier ◽

Tonia Woodberry ◽

Marcus Altfeld ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Chemokine Receptor ◽

Cd8 T Cells ◽

Molecular Mechanisms ◽

Cell Subset ◽

Cd8 T Cell ◽

Effector Memory ◽

Immune System Activation ◽

Hiv 1

CD8 T cells play a key role in host defense against intracellular pathogens. Efficient migration of these cells into sites of infection is therefore intimately linked to their effector function. The molecular mechanisms that control CD8 T-cell trafficking into sites of infection and inflammation are not well understood, but the chemokine/chemokine receptor system is thought to orchestrate this process. Here we systematically examined the chemokine receptor profile expressed on human CD8 T cells. Surprisingly, we found that CXC chemokine receptor 1 (CXCR1), the predominant neutrophil chemokine receptor, defined a novel interleukin-8/CXC ligand 8 (IL-8/CXCL8)–responsive CD8 T-cell subset that was enriched in perforin, granzyme B, and interferon-γ (IFNγ), and had high cytotoxic potential. CXCR1 expression was down-regulated by antigen stimulation both in vitro and in vivo, suggesting antigen-dependent shaping of the migratory characteristics of CD8 T cells. On virus-specific CD8 T cells from persons with a history of Epstein-Barr virus (EBV) and influenza infection, CXCR1 expression was restricted to terminally differentiated effector memory cells. In HIV-1 infection, CXCR1-expressing HIV-1–specific CD8 T cells were present only in persons who were able to control HIV-1 replication during structured treatment interruptions. Thus, CXCR1 identifies a subset of CD8 T cells poised for immediate cytotoxicity and early recruitment into sites of innate immune system activation.

Download Full-text

Degranulation plays an essential part in regulating cell surface expression of Fas ligand in T cells and natural killer cells

Nature Medicine ◽

10.1038/4779 ◽

1999 ◽

Vol 5 (1) ◽

pp. 90-96 ◽

Cited By ~ 259

Author(s):

G. Bossi ◽

G.M. Griffiths

Keyword(s):

T Cells ◽

Natural Killer Cells ◽

Cell Surface ◽

Natural Killer ◽

Fas Ligand ◽

Surface Expression ◽

Cell Surface Expression ◽

Killer Cells

Download Full-text

Bystander hyperactivation of preimmune CD8+ T cells in chronic HCV patients

eLife ◽

10.7554/elife.07916 ◽

2015 ◽

Vol 4 ◽

Cited By ~ 29

Author(s):

Cécile Alanio ◽

Francesco Nicoli ◽

Philippe Sultanik ◽

Tobias Flecken ◽

Brieuc Perot ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Chronic Infection ◽

Surface Expression ◽

Cross Sectional Study ◽

Negative Regulator ◽

Tcr Signaling ◽

Cell Repertoire ◽

Cross Sectional ◽

Chronic Hcv

Chronic infection perturbs immune homeostasis. While prior studies have reported dysregulation of effector and memory cells, little is known about the effects on naïve T cell populations. We performed a cross-sectional study of chronic hepatitis C (cHCV) patients using tetramer-associated magnetic enrichment to study antigen-specific inexperienced CD8+ T cells (i.e., tumor or unrelated virus-specific populations in tumor-free and sero-negative individuals). cHCV showed normal precursor frequencies, but increased proportions of memory-phenotype inexperienced cells, as compared to healthy donors or cured HCV patients. These observations could be explained by low surface expression of CD5, a negative regulator of TCR signaling. Accordingly, we demonstrated TCR hyperactivation and generation of potent CD8+ T cell responses from the altered T cell repertoire of cHCV patients. In sum, we provide the first evidence that naïve CD8+ T cells are dysregulated during cHCV infection, and establish a new mechanism of immune perturbation secondary to chronic infection.

Download Full-text

Costimulation of T cells by B7-H2, a B7-like molecule that binds ICOS

Blood ◽

10.1182/blood.v96.8.2808 ◽

2000 ◽

Vol 96 (8) ◽

pp. 2808-2813 ◽

Cited By ~ 180

Author(s):

Shengdian Wang ◽

Gefeng Zhu ◽

Andrei I. Chapoval ◽

Haidong Dong ◽

Koji Tamada ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Chinese Hamster Ovary Cells ◽

Chinese Hamster ◽

Surface Expression ◽

Cell Surface Expression ◽

Ovary Cells ◽

Inducible Costimulator ◽

Immature Dendritic Cells ◽

Dose Dependent

Abstract This report describes a new human B7-like gene designatedB7-H2. Cell surface expression of B7-H2 protein is detected in monocyte-derived immature dendritic cells. Soluble B7-H2 and immunoglobulin (Ig) fusion protein, B7-H2Ig, binds activated but not resting T cells and the binding is abrogated by inducible costimulator Ig (ICOSIg), but not CTLA4Ig. In addition, ICOSIg stains Chinese hamster ovary cells transfected with B7-H2 gene. By suboptimal cross-linking of CD3, costimulation of T-cell proliferation by B7-H2Ig is dose-dependent and correlates with secretion of interleukin (IL)-2, whereas optimal CD3 ligation preferentially stimulates IL-10 production. The results indicate that B7-H2 is a putative ligand for the ICOS T-cell molecule.

Download Full-text

Chlorotoxin-directed CAR T cells for specific and effective targeting of glioblastoma

Science Translational Medicine ◽

10.1126/scitranslmed.aaw2672 ◽

2020 ◽

Vol 12 (533) ◽

pp. eaaw2672 ◽

Cited By ~ 15

Author(s):

Dongrui Wang ◽

Renate Starr ◽

Wen-Chung Chang ◽

Brenda Aguilar ◽

Darya Alizadeh ◽

...

Keyword(s):

T Cells ◽

Tumor Regression ◽

Surface Expression ◽

Matrix Metalloproteinase 2 ◽

Cell Surface Expression ◽

Binding Potential ◽

Car T Cells ◽

Car T ◽

Peptide Toxin ◽

Effector Activity

Although chimeric antigen receptor (CAR) T cells have demonstrated signs of antitumor activity against glioblastoma (GBM), tumor heterogeneity remains a critical challenge. To achieve broader and more effective GBM targeting, we developed a peptide-bearing CAR exploiting the GBM-binding potential of chlorotoxin (CLTX). We find that CLTX peptide binds a great proportion of tumors and constituent tumor cells. CAR T cells using CLTX as the targeting domain (CLTX-CAR T cells) mediate potent anti-GBM activity and efficiently target tumors lacking expression of other GBM-associated antigens. Treatment with CLTX-CAR T cells resulted in tumor regression in orthotopic xenograft GBM tumor models. CLTX-CAR T cells do not exhibit observable off-target effector activity against normal cells or after adoptive transfer into mice. Effective targeting by CLTX-CAR T cells requires cell surface expression of matrix metalloproteinase–2. Our results pioneer a peptide toxin in CAR design, expanding the repertoire of tumor-selective CAR T cells with the potential to reduce antigen escape.

Download Full-text

Different structural constraints for recognition of mouse H-2Kd and -Kk antigens by alloimmune cytolytic T lymphocytes.

Journal of Experimental Medicine ◽

10.1084/jem.164.6.1823 ◽

1986 ◽

Vol 164 (6) ◽

pp. 1823-1834 ◽

Cited By ~ 4

Author(s):

J Schøller ◽

R Shimonkevitz ◽

H R MacDonald ◽

S Kvist

Keyword(s):

T Cells ◽

Amino Acid ◽

T Lymphocytes ◽

Surface Expression ◽

Cell Surface Expression ◽

Structural Constraints ◽

The Novel ◽

Different Types ◽

Target Molecules ◽

Hybrid Genes

We have constructed a new series of hybrid genes among the H-2Kd,-Kk, and -Kb. The site of recombination occurs in the third exon, encoding the alpha 2 domain, and divides this domain into two parts, alpha 2A and alpha 2B. The novel genes differ only in the COOH-terminal half of the alpha 2 domain, i.e., the alpha 2B region. This region, comprising residues 142-182, contains a limited number of amino acid differences between the three alleles. The hybrid genes have been introduced into 1T 22-6 cells (H-2q), and cell surface expression of hybrid antigens was verified. Cells expressing different types of hybrid antigens have been examined for their susceptibility to lysis by cytotoxic T lymphocytes directed either against the H-2Kd antigen or the H-2Kk antigen. Our results show that the alpha 1 and alpha 2A domains of the H-2Kk antigen can constitute target molecules for alloimmune anti-Kk T cells, whereas the alpha 2B region, when exchanged for Kd or Kb sequences, plays only a limited role. In contrast, the alpha 1 and alpha 2A domains of Kd are not sufficient to be recognized by alloimmune anti-Kd T cells. In this instance, the alpha 2B domain seems to play an essential role. This region has undergone several amino acid substitutions involving charged residues.

Download Full-text