scholarly journals Analysis of T-Cell Receptor Vβ Gene Repertoires after Immune Stimulation and in Malignancy by Use of Padlock Probes and Microarrays

2005 ◽  
Vol 51 (4) ◽  
pp. 768-775 ◽  
Author(s):  
Johan Banér ◽  
Per Marits ◽  
Mats Nilsson ◽  
Ola Winqvist ◽  
Ulf Landegren
Keyword(s):  
T Cell ◽  
Lymph Nodes ◽  
Peripheral Blood ◽  
Tumor Infiltrating Lymphocytes ◽  
Staphylococcal Enterotoxin ◽  
Staphylococcal Enterotoxin B ◽  
Padlock Probes ◽  
Enterotoxin B ◽  
Infiltrating Lymphocytes ◽  
Draining Lymph Nodes

Abstract Background: Detection of expanded T-cell clones, identified by their receptor (TCR) repertoires, can assist diagnosis and guide therapy in infectious, inflammatory, and autoimmune conditions as well as in tumor immunotherapy. Analysis of tumor-infiltrating lymphocytes often reveals preferential use of one or a few TCR Vβ genes, compared with peripheral blood, indicative of a clonal response against tumor antigens. Methods: To simultaneously measure the relative expression of all Vβ gene families, we combined highly specific and sensitive oligonucleotide reagents, called padlock probes, with a microarray read-out format. T-Cell cDNA was combined with a pool of Vβ subfamily-specific padlock probes. Reacted probes were selectively amplified and the products hybridized to a microarray, from which the Vβ subfamily distribution in each sample could be determined relative to a control sample. Results: In lymphocytes stimulated with the superantigen staphylococcal enterotoxin B, we detected expansions at the mRNA level of TCR subfamilies previously shown to respond to staphylococcal enterotoxin B. Expansions of the same Vβ families could also be detected by flow cytometry. In samples from two bladder cancer patients, we detected predominant representations of specific Vβ subfamilies in both tumor-infiltrating lymphocytes and in the draining lymph nodes, but not in non-tumor-draining lymph nodes or peripheral blood. Several expression profiles from draining lymph nodes in patients with malignant melanoma were divergent from profiles seen in non-tumor-draining lymph nodes. Conclusion: Padlock probe-based parallel analysis of TCR Vβ gene distributions provides an efficient method for screening multiple samples for T-cell clonal expansions with reduced labor and time of analysis compared with traditional methods.

scholarly journals Anti-Vβ8 antibodies induce and maintain staphylococcal enterotoxin B-triggered Vβ8+ T cell anergy

1999 ◽  
Vol 29 (2) ◽  
pp. 437-445 ◽  
Author(s):  
Luiz Stark Aroeira ◽  
Concepción G. Mouton ◽  
José L. Toran ◽  
Elizabeth Sally Ward ◽  
Carlos Martínez-A.
Keyword(s):  
T Cell ◽  
Staphylococcal Enterotoxin ◽  
Staphylococcal Enterotoxin B ◽  
T Cell Anergy ◽  
Enterotoxin B ◽  
Cell Anergy

scholarly journals CD4 + T‐cell‐mediated cytotoxicity against staphylococcal enterotoxin B‐pulsed synovial cells

Immunology ◽  
1998 ◽  
Vol 95 (1) ◽  
pp. 38-46 ◽  
Author(s):  
KAWAKAMI ◽  
MATSUOKA ◽  
TSUBOI ◽  
URAYAMA ◽  
NAKASHIMA ◽  
...  
Keyword(s):  
T Cell ◽  
Staphylococcal Enterotoxin ◽  
Cd4 T Cell ◽  
Staphylococcal Enterotoxin B ◽  
Synovial Cells ◽  
Enterotoxin B

Staphylococcal enterotoxin B induces fever, brain c-Fos expression, and serum corticosterone in rats

2001 ◽  
Vol 280 (5) ◽  
pp. R1434-R1439 ◽  
Author(s):  
Lisa E. Goehler ◽  
Ron P. A. Gaykema ◽  
Michael K. Hansen ◽  
Jayme L. Kleiner ◽  
Steven F. Maier ◽  
...  
Keyword(s):  
T Cell ◽  
Host Defense ◽  
Intraperitoneal Injection ◽  
Defense Responses ◽  
Plasma Corticosterone ◽  
Significant Rise ◽  
Staphylococcal Enterotoxin ◽  
Staphylococcal Enterotoxin B ◽  
Enterotoxin B ◽  
Fos Expression

The paraventricular nucleus of the hypothalamus (PVH) occupies a pivotal point within the network of brain nuclei coordinating critical host-defense responses. In mice, T cell-dependent immune stimuli, including the bacterial superantigen staphylococcal enterotoxin B (SEB), can activate the PVH. To determine whether T cell-dependent immune stimuli activate the PVH in rats, we assessed plasma corticosterone (Cort) levels, fever responses, and c-Fos expression in the PVH in animals treated with intraperitoneal injections of SEB. In animals with previously implanted abdominal thermisters, intraperitoneal injection of 1 mg/kg SEB resulted in a significant rise in body temperature, with a latency of 3.5–4 h. In separate animals, intraperitoneal injection of 1 mg/kg SEB resulted in a significant elevation of plasma Cort and induced c-Fos expression in parvocellular neurons within the PVH. These results support the idea that T cell-dependent immune stimuli activate brain pathways mediating host-defense responses such as fever and neuroendocrine changes.

T-cell receptor V-gene usage in neoplasms of the central nervous system

1993 ◽  
Vol 78 (4) ◽  
pp. 630-637 ◽  
Author(s):  
Adrian Merlo ◽  
Luis Filgueira ◽  
Markus Zuber ◽  
Antonio Juretic ◽  
Felix Harder ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Tumor Infiltrating Lymphocytes ◽  
Cell Receptor ◽  
Gene Products ◽  
Peripheral Blood Mononuclear ◽  
Infiltrating Lymphocytes

✓ The use of tumor-infiltrating lymphocytes in the treatment of central nervous system (CNS) neoplasms has met with serious obstacles due to difficulty of culture and poor characterization. Since in other tumors the therapeutic effects of tumor-infiltrating lymphocytes have been shown to rely on T-cell receptor engagement, the authors addressed the question as to whether expression of T-cell receptor variable (V) domains in cultured tumor-infiltrating lymphocytes from CNS is different from that of autologous cultured peripheral blood mononuclear cells. Infiltrating lymphocytes from CNS neoplasms, including primary malignancies, metastatic cancers, and meningiomas, were cultured in the presence of interleukin-2 and anti-CD3 monoclonal antibodies (MoAb's) in order to obtain optimum growth of T cells. Autologous peripheral blood mononuclear cells from the same patients were similarly cultured. After 4 to 5 weeks of culture, 97.3% ± 2.6% (mean ± standard deviation) of the resulting cell populations were CD3-positive lymphocytes. The expression of T-cell receptor V domains was then studied by using a panel of 12 MoAb recognizing gene products from T-cell receptor V-α 2, V-β 5, 6, 8, and 12, V-γ 4 and 9 families, and from two subfamilies of V-δ 2. Remarkably, in over 70% of all paired measurements, percentages of T cells expressing discrete T-cell receptor V-gene products were found to be virtually identical in tumor- and peripheral blood-derived cultured cell populations, with differences never exceeding 1%. In contrast, a different expression of individual V-gene products, concerning both α/β and γ/δ T-cell receptors, could be detected between cultured tumor-infiltrating lymphocytes and autologous peripheral blood-derived T lymphocytes in seven of 12 patients. In two cases, significant differences between the two populations were also observed in the proliferative responses obtained upon stimulation with staphylococcal enterotoxins that trigger defined V-β T-cell receptors. Altogether, these data suggest that the T-cell receptor repertoire of cultured tumor-infiltrating lymphocytes from CNS tumors, suitable for use in adoptive immunotherapies, differs from that of autologous cultured peripheral blood mononuclear cells.

scholarly journals Induction of unresponsiveness and impaired T cell expansion by staphylococcal enterotoxin B in CD28-deficient mice.

1996 ◽  
Vol 183 (6) ◽  
pp. 2481-2488 ◽  
Author(s):  
H W Mittrücker ◽  
A Shahinian ◽  
D Bouchard ◽  
T M Kündig ◽  
T W Mak
Keyword(s):  
T Cells ◽  
T Cell ◽  
Staphylococcal Enterotoxin ◽  
Staphylococcal Enterotoxin B ◽  
Rapid Induction ◽  
Cd28 Costimulation ◽  
Enterotoxin B ◽  
Deficient Mice

We used CD28-deficient mice to analyze the importance of CD28 costimulation for the response against Staphylococcal enterotoxin B (SEB) in vivo. CD28 was necessary for the strong expansion of V beta 8+ T cells, but not for deletion. The lack of expansion was not due to a failure of SEB to activate V beta 8+ T cells, as V beta 8+ T cells from both CD28-/- and CD28+/+ mice showed similar phenotypic changes within the first 24 h after SEB injection and cell cycle analysis showed that an equal percentage of V beta 8+ T cells started to proliferate. However, the phenotype and the state of proliferation of V beta 8+ T cells was different at later time points. Furthermore, in CD28-/- mice injection with SEB led to rapid induction of unresponsiveness in SEB responsive T cells, indicated by a drastic reduction of proliferation after secondary SEB stimulation in vitro. Unresponsiveness could also be demonstrated in vivo, as CD28-/- mice produced only marginal amounts of TNF alpha after rechallenge with SEB. In addition CD28-/- mice were protected against a lethal toxic shock induced by a second injection with SEB. Our results indicate that CD28 costimulation is crucial for the T cell-mediated toxicity of SEB and demonstrate that T cell stimulation in the absence of CD28 costimulation induces unresponsiveness in vivo.

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