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PubMedDate not supplied

T-cell receptor V beta gene usage of human cytotoxic T-cell clones obtained from gastric cancer patients.

Nine T-cell clones have been established from tumor-infiltrating lymphocytes (TIL) isolated from ascitic fluid of a gastric cancer patient. Five of them retained cytotoxicity against autologous tumor cells (AuTu), and were all CD4+. Each clone had different usage of T-cell receptor (TCR) V beta gene as assessed by Southern blot analysis. Using AuTu and two allogeneic gastric cancer cell lines as targets, we selected three clones with unique cytotoxic properties. Two of these clones (Clone 1 and 2) preferentially lysed AuTu, but showed no or marginal cytotoxicity against allogeneic gastric cancer cells, and one clone (Clone 7) showed appreciable cytotoxicity against AuTu and allogeneic gastric cancer cells. In the detailed analysis of TCRV beta gene usage, Clone 1, 2, and 7 expressed V beta 13.1/D beta 1/J beta 1.5/C beta 1, V beta 3/D beta 2/J beta 2.4/C beta 2, and V beta 9/D beta 1/J beta 1.4/C beta 1, respectively, and the primary structures of the three TCRVb genes did not share any common features, neither in the sizes of their complementarity determining region 3 (CDR3) nor in their amino acid compositions. Interestingly, PBL of the same patient expressed CDR3 identical to that of Clone 2 and 7, but not that of Clone 1. CDR3 identical to that of Clone 2 and 7 were also detected in TIL of other gastric cancer patients. These results show that some AuTu-specific CTL included in TIL are circulating in peripheral blood, and that the CDR3 identical to that of the CTL is expressed extensively in TIL among different gastric cancer patients. Screening of the expression of the CDR3 in other gastric cancer patients is recommended to develop an immuno-therapy of gastric cancer based on antigenic peptide.

Adenocarcinoma↗

PubMedDate not supplied

Anti-endothelial cell antibodies in Behçet's disease.

OBJECTIVE: Circulating antibodies that bind to human endothelial cells cultured in vitro have been detected in a variety of diseases, including Behçet's disease. In this disorder the reported prevalence of AECA has varied widely. One likely source of variability is the ELISA assay itself, in which differing conditions and reagents have been used in different reports. METHODS: We have re-examined the frequency of AECA in 132 Turkish Behçet's patients and 50 healthy Turkish controls, comparing several different methods of preparing the target endothelial cells. Human umbilical vein endothelial cells (HUVEC) were used either: 1) fresh and non-treated, 2) fixed, or 3) TNF alpha-stimulated. All stages of the procedures were performed at room temperature. RESULTS: In Behçet's patients, using fresh, non-treated HUVEC, 17 of 130 (13.1%) and 9 of 132 (6.8%) sera were positive for IgG- and IgM-AECA, respectively. However, among 50 normal controls, 2 (4.0%) had IgG-positive and 4 (8.0%) had IgM-positive ELISAs under the same conditions. The difference in the frequency of positives between patients and controls was not statistically significant. Fixed HUVEC and TNF alpha-treated HUVEC gave similar results as well. When group means were examined, only the mean for IgG-AECA determined with TNF alpha-stimulated HUVEC reached statistical significance. CONCLUSION: The discrepancy between our data and earlier reports in the literature probably reflects the methodological differences alluded to, and highlights the difficulties in interpreting ELISA assays for AECA.

Adult↗