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目的分析重组人软骨抗原凝集原 (rG1 )体外诱导建立的 1 1株rG1特异性T细胞株的生物学特性。方法采用直接标记免疫荧光法对rG1特异性T细胞株进行表型分析 ;采用QuantikineELISAKits定量检测细胞培养上清液中的IFN -γ与IL - 4;采用RT -PCR方法分析其BV的限制性取用。 结果所建细胞株均表现为CD3阳性 (98.6 %± 1 .5 % ) ,大部分细胞株 (9/ 1 1 )为CD4阳性 (97.1 %± 2 .5 % ) ,而其余 2株为CD4和CD8混合表达 ,其中CD4阳性为 82 .95 % ,CD8阳性为 36 .5 0 %。 8株分泌IFN -γ(5 44± 390 pg/ml) ,而IL - 4阴性 ,为Thl细胞 ;1株同时分泌IFN -γ和IL - 4,为Th0细胞 ;2株细胞未测出IFN -γ和IL - 4分泌 ;无Th2细胞株出现。所有T细胞株均选择性取用αβ(97.4%± 2 .4% )TCR受体。 结论所建立的rG1特异性T细胞株经表面鉴定为纯度很高的CD3阳性T细胞 ,多表现为CD4阳性 ,细胞因子分泌类型多属Th1T细胞。
Objective To analyze the biological characteristics of 11 rG1-specific T cell lines induced by recombinant human chondrogenic antigen agglutinogens (rG1) in vitro. Methods The direct immunofluorescence assay was used to analyze the phenotype of rG1 - specific T cell lines. Quantikine ELISAKits were used to detect IFN - γ and IL - 4 in cell culture supernatants. The restriction of BV was analyzed by RT - PCR use. Results Most of the cell lines were positive for CD3 (98.6% ± 1.5%), most of the cells were CD4 (97.1% ± 2.5%), while the remaining 2 were CD4 CD8 mixed expression, CD4 positive for 82.95%, CD8 positive for 36.500%. 8 strains secreted IFN - γ (5 44 ± 390 pg / ml), but IL - 4 was negative, which was Th1 cells. One strain secreted IFN - γ and IL - 4 at the same time and was Th0 cells. γ and IL - 4 secretion; no Th2 cell lines appear. All T cell lines selectively take αβ (97.4% ± 2 .4%) TCR receptors. Conclusion The established rG1-specific T cell lines were identified as highly pure CD3-positive T cells, mostly CD4 positive, and cytokine secretion mostly Th1 T cells.