特异性细胞毒性T细胞检测方法的建立及其在重组痘苗病毒活疫苗细胞免疫研究中的应用

报道了用ＣＢＡ／Ｊ小鼠、Ｌ９２９细胞及ＭＴＴ比色法建立的特异性细胞毒性Ｔ细胞（ＣＴＬ）的检测方法及其在重组痘苗病毒活疫苗细胞免疫研究中的应用。通过检测痘菌病毒特异性初发ＣＴＬ的动态变化及其二次反应ＣＴＬ，表明，本法能较好地检测初发及二次反应ＣＴＬ水平的变化。用本法检测了不同启动子控制下的表达Ｅｐｓｔｅｉｎ－Ｂａｒｓ（ＥＢ）病毒膜抗原（ＥＢＶ－ＭＡ）的重组痘菌病毒ＥＢＶ－ＭＡ特异性ＣＴＬ、以及与表达人白细胞介素－２的重组痘苗病毒联合免疫时的ＥＢＶ－ＭＡ特异性ＣＴＬ。结果表明，Ｐ７５００启动子表达的ＥＢＶ－ＭＡ比Ｐ１１０００启动子表达的ＥＢＶ－ＭＡ诱导的ＥＢＶ－ＭＡ特异性ＣＴＬ高，但无显著性差异。当上述重组痘苗病毒与表达人白细胞介素－２的重组痘苗病毒联合免疫时，均能使二者的ＥＢＶ－ＭＡ特异性ＣＴＬ增高，但增高幅度本身及其二者之间均无显著性差异。

The Establishment of the Method for Detection of Specific Cytotoxic T Lymphocytes and Its Application in the Cellular Immunity studies of Recombinant Vaccinia Live Vaccines

This paper describes the method for detection of specific cytotoxic T lymphocytes (CTL) with CBA/J mouse, L929cell and MTT colorimetric assay and it application in the cellular immunity studies of recombinant live vaccines, This method detected the Kinetic Changes of primary CTLs and differences of secondarily stimulated CTLs well. With this method, the EBV-MA specific CTLs induced by recombinant vaccinia viruses expressing EBV-MA under control of different promoters.rVV-EBV-MA(P7500)and rVV-EBV-MA (P11000), as well as the EBV-MA specific CTLs induced by the above two recombinant viruses in combination with recombinant vaccinia virus expressing human Interleukin-2 were detected. The results indicate that rVV- EBV- MA (P7500) induced slightly higher EBV- MA specific CTL than rVV-EBV-MA (P11000), however, no statistical difference was found. Obvious enhancement of EBV-MA specific CTL was observed when immunized by the above two recombinant vaccinia viruses in combination with rVV-IL-2, but the enhancement was not statistically significant.