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目的 :研究了经Cop I治疗后的多发性硬化 (MS)患者外周血淋巴细胞 (PBMC)对Cop I及碱性髓鞘蛋白 (MBP)免疫的反应性。方法 :用半有限稀释微量培养技术分析经Cop I治疗的MS患者的PBMC对Cop I和MBP诱导的特异性T细胞株的反应频率。用ELISA方法检测T细胞株分泌的细胞因子类型。用免疫荧光检测了特异性Cop I细胞株的表型。结果 :15位经Cop I治疗后的MS患者PBMC中的Cop I/MBP特异性T细胞克隆反应频率高达 84 0 % ,而在 9位非MS患者对照组中 ,其T细胞对Cop I/MBP反应的频率仅为 5 4%。用ELISA方法检测发现体外Cop I/MBP诱导的特异性T细胞株株分泌IL 5和IL 13 ,为Th2类型的细胞因子。所有的细胞株均表现为CD3阳性 ,大部分高表达CD8细胞并发现有CD3/CD5 6双阳性的NKT细胞。结论 :上述结果提示多肽疫苗Cop I能诱导MS患者特异性免疫反应 ,以诱导CD8阳性T细胞和NKT为特点 ,并以正向调节Th2类型为主的免疫活性细胞
AIM: To investigate the responsiveness of peripheral blood lymphocytes (PBMCs) to Cop I and basic myelin protein (MBP) immunization in patients with multiple sclerosis (MS) after Cop I treatment. METHODS: The semi-limited dilution microplate technique was used to analyze the frequency of PBMC responses to Cop I and MBP-induced specific T cell lines in Cop I -treated MS patients. The type of cytokines secreted by T cell lines was detected by ELISA. The phenotypes of specific Cop I cell lines were examined by immunofluorescence. RESULTS: The Cop I / MBP-specific T cell clones in 15 PBMCs treated with Cop I had a high frequency of 84 0%. However, in 9 non-MS patients, their T cell responses to Cop I / MBP The reaction frequency is only 54%. It was found by ELISA that the secreted IL-5 and IL-13 secreted by Cop I / MBP-induced T cell lines in vitro were Th2 type cytokines. All cell lines showed positive for CD3, the majority of CD8 cells were highly expressed and CD3 / CD5 6 double positive NKT cells were found. CONCLUSION: The above results suggest that the peptide vaccine Cop I can induce specific immune response in MS patients, which is characterized by induction of CD8 + T cells and NKT, and positively regulate Th2 type-based immunocompetent cells