建立猪MHCⅠ类抗原特异性人T细胞系的探讨

Establishment of human T lymphocyte lines specific to swine leukocyte antigen class Ⅰ-P1 molecules

目的 建立间接识别中国版纳猪SLAⅠ类P1分子的特异性人T细胞系的方法 ,以进一步研究SLAⅠ类分子的间接识别在猪→人异种细胞性排斥中的作用。方法 以E·coli中表达并纯化的SLAⅠ类分子P1为抗原 ,体外反复刺激健康人PBMC ,3H TdR掺入法筛选特异性增殖的T细胞 ,FACS作表型初步分析。结果 初步测定健康人外周血版纳猪SLAⅠ类P1分子特异性T细胞反应频率约 6 .67× 1 0 - 7,所建 4个T细胞系表型均为TCRαβ+ ,其中 3株以CD4+ 为主 ,1株以CD8+ 为主。结论 利用E .coli表达的纯蛋白抗原可建立间接识别版纳猪SLAⅠ类P1分子的特异性人T细胞系。

Objective To establish human specific T cell lines which indirectly recognize Chinese Banna Pig SLA class Ⅰ P1 molecules so as to identify the role of SLA class Ⅰ molecules in acute cellular xeno responses between swine and human. Methods Using purified recombinant Banna Pig P1 protein produced in E.coli as antigen to stimulate human PBMCs repeatedly. The T cells specific to P1 antigen were selected by 3H TdR incorporation. Phenotype analysis was assayed by FACS. Results The frequency of T cells specific to SLA P1 molecule was preliminarily 6.67×10 -7 in healthy donor PBMC. All of four specific T cell lines were TCRαβ positive .Three of them were predominantly CD4 positive, the other was predominantly CD8 positive.Conclusions Human specific T cell lines that indirectly recognizes SLA class Ⅰ are established by using purified recombinant protein P1 as antigen.