Vβ-Specific Stimulation of Human T Cells by Staphylococcal Toxins

Kappler, John W.; Kotzin, Brian L.; Herron, L R; Gelfand, Erwin W.; Bigler, Robert D.; Boylston, Arthur W.; Carrel, Stefan; Posnett, David N.; Choi, Yongwon; Marrack, Philippa

doi:10.1126/science.2524876

Cited by 677 publications

(440 citation statements)

References 31 publications

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“…Recently, we have shown that provocative stimuli such as the superantigen SEB, which binds specifically to the Vp region of the T-cell receptor (2,15), were capable of stimulating distinct Vp subsets of T cells (Maino et al, in preparation). Multiparameter flow cytometry employing FITC-labeled antibodies to specific Vp T-cell receptor antigens indicated that CD69 expression in response to SEB was consistent with the expected specificities of Vp subsets reported for SEB ( 11). For example, SEB stimulated CD69 expression on Vp6+ subset of T cells but did not stimulate Vp8' T cells.…”

Section: Antigen-and Superantigen-induced T-cell Responsessupporting

confidence: 72%

Rapid flow cytometric method for measuring lymphocyte subset activation

1995

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Standard in vitro methods for assessing T-cell activation have typically measured either the proliferative responses of peripheral blood mononuclear cell (PBMC) cultures to various provocative stimuli employing tritiated thymidine incorporation or the secretion of specific cytokines from activated cells. These bulk assay methods suffer the drawback of being lengthy assays, and, in addition, they do not provide information about functional responses of individual lymphocyte subsets. This report describes a three-color flow cytometric method for the rapid analysis (4 hours) of individual T-cell subsets in whole blood responding to various provocative stimuli, including pokeweed mitogen, the comitogenic monoclonal antibodies (mAbs) CD2/CD2R, the superantigen staphylococcal enterotoxin B (SEB), and the speciac antigen Candlda albicans. After 4 hours, CD69 expression in response to CD2/ CDZR paralleled thymidine incorporation measured after 72 hours. Variations in the proportions of CD4+ and CD4-T cells expressing CD69 were observed with Merent stimuli. These observations demonstrate the potential of multiparameter flow cytometry for the investigation of functional responses of individual T-cell subsets to a variety of stimuli in whole blood. o 1995 wiley-Liss, Inc.

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Section: Antigen-and Superantigen-induced T-cell Responsessupporting

confidence: 72%

Rapid flow cytometric method for measuring lymphocyte subset activation

1995

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“…However, the extent of proliferation measured in heterogeneous responder cell populations does not allow precise determination of the participation of individual subpopulations or the proportion of cells contributing to overall thymidine incorporation (3). In addition, the quality of conventional cytotoxicity assays frequently suffers from the spontaneous (i.e., false positive) release of chromium or thymidine andlor from the requirement for an incubation period longer than [6][7][8] h with certain target cells. Furthermore, the experimental evaluation of activationinduced cell death, also referred to as apoptosis or programmed cell death, requires enumeration of the number of viable and dead cells in order to quantify cell death of the responding lymphocytes (7,12).…”

Section: Key Terms: Quantitative Flow Cytometry T-cell Activation Ymentioning

confidence: 99%

Rapid quantification of lymphocyte subsets in heterogeneous cell populations by flow cytometry

1994

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Determination of the number of viable cells or quantification of lymphocyte subsets in heterogeneous cell populations is critically important for cytotoxicity assays, apoptosis assays, or the analysis of differential activation of T‐cell subsets by distinct stimuli. In this report, we describe a rapid flow cytometry method termed Standard Cell Dilution Analysis (SCDA) specifically to quantify any subset of phenotypically definable, viable cells in heterogeneous populations using a FACScan flow cytometer. This method combines: (1) specific detection of lymphocyte subsets by phycoerythrin‐conjugated monoclonal antibodies, (2) electronic exclusion of dead cells or cell debris by propidium‐iodide staining and gating on forward vs. sidescatter, respectively, and (3) admixture of a known amount of fixed, fluorescein isothiocyanate stained cells immediately before analysis as a constant parameter to allow for calculation of cell quantity. We have used SCDA to analyze the in vitro growth characteristics of various human T‐lymphocyte subpopulations in response to different activation stimuli. © 1994 Wiley‐Liss, Inc.

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“…There are reports that T cells bearing particular TCR V{3 elements can be stimulated either by minor lymphocyte stimulating (MIs) determinants or by bacterial toxins such as toxic shock syndrome toxin (TSST) and staphylococcal enterotoxin (5,6). The mode of stimulation of T cells by these toxins may be similar to that of the MIs determinants, and both MIs determinants and bacterial toxins have been termed "superantigens."…”

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confidence: 99%

Restricted junctional usage of T cell receptor Vβ2 and Vβ13 genes, which are overrepresented on infiltrating T cells in the lips of patients with sjögren's syndrome

Yonaha

Sumida

Maeda

et al. 1992

Arthritis & Rheumatism

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Objective. To analyze the clonality of T cell receptor (TCR) Vβ2—and Vβ13—positive T cells, which are predominantly expressed in the lips of patients with Sjögren's syndrome (SS).Methods. The junctional sequences of complementary DNA clones encoding TCR Vβ2 and Vβ13 genes were determined by the polymerase chain reaction. Forty‐one Vβ2 and 45 Vβ13 clones established from the lips of 3 SS patients were sequenced.Results. The Vβ2/Jβ2.3 pair was enriched in 2 of the 3 patients (44% and 46% of the clones, respectively), and the Vβ13/Jβ2.1 sequence was dominant in 2 of the 3 (23% and 45%). These pairs were not used preferentially in peripheral blood lymphocytes from the same patients.Conclusion. Infiltrating Vβ2‐ and Vβ13‐positive T cells from the lips of all 3 patients with SS were polyclonal, but the junctional usage of cells from 2 lip samples was restricted, compared with cells from peripheral blood. This suggests that not all expanded cells from the lips of SS patients are stimulated by superantigens.

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Vβ-Specific Stimulation of Human T Cells by Staphylococcal Toxins

Cited by 677 publications

References 31 publications

Rapid flow cytometric method for measuring lymphocyte subset activation

Rapid flow cytometric method for measuring lymphocyte subset activation

Rapid quantification of lymphocyte subsets in heterogeneous cell populations by flow cytometry

Restricted junctional usage of T cell receptor Vβ2 and Vβ13 genes, which are overrepresented on infiltrating T cells in the lips of patients with sjögren's syndrome

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