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目的观察1型糖尿病反义肽噬菌体疫苗诱导CD8+T细胞对病理性CD4+T细胞的抑制作用。方法用1型糖尿病反义肽噬菌体疫苗免疫非肥胖型糖尿病(NOD)小鼠,并同时设立噬菌体空载体免疫组和未免疫空白对照组。于初次免疫后第20周,检测各组小鼠血糖;磁珠法分离免疫小鼠CD8+T细胞,并用合成反义肽及IL-2诱导刺激作为效应细胞;分离噬菌体空载体免疫组和空白对照组小鼠CD4+T细胞,用合成正义肽及IL-2诱导刺激作为靶细胞。将效应细胞与靶细胞按不同比例混合,以乳酸脱氢酶(LDH)释放法检测CTL的杀伤活性。结果初次免疫后第20周,1型糖尿病反义肽噬菌体疫苗免疫组小鼠血糖水平保持正常,而另外2组小鼠血糖均高于正常水平。1型糖尿病反义肽噬菌体疫苗免疫组诱导的CD8+T细胞作效应细胞,当效靶比为100∶1时,对噬菌体空载体免疫组CD4+T细胞的杀伤效率最高,达47.95%±11.30%,而噬菌体空载体免疫组诱导的小鼠CD8+T细胞对空白对照组的CD4+T细胞无杀伤作用。结论1型糖尿病反义肽噬菌体疫苗能够诱导CD8+T细胞抑制病理性CD4+T细胞。
Objective To observe the inhibitory effect of CD8 + T cells on pathological CD4 + T cells induced by type 1 diabetes antisense peptide phage vaccine. Methods Non-obese diabetic (NOD) mice were immunized with the type 1 diabetes antisense peptide phage vaccine and the phage empty vector and non-immunized blank control groups were established. At the 20th week after the initial immunization, the blood glucose of each group was detected. The CD8 + T cells of immunized mice were isolated by magnetic beads method and stimulated by synthetic antisense peptide and IL-2 as effector cells. CD4 + T cells of control mice were stimulated with synthetic sense peptide and IL-2 as target cells. The effector cells and target cells were mixed in different proportions, and the cytotoxic activity of CTLs was detected by lactate dehydrogenase (LDH) release assay. Results At the 20th week after the first immunization, the blood glucose level of the mice immunized with the antisense peptide phage type 1 diabetes vaccine remained normal while the blood glucose of the other two groups of mice was higher than the normal level. The CD8 + T cells induced by the type 1 diabetes antisense peptide phage vaccine group were the effector cells with the highest efficiency of 47.95% ± 11.30 against the CD4 (superscript +) T cells of the phage empty vector immunized group when the target ratio was 100: 1 %, While the murine CD8 + T cells induced by the phage empty vector immunity group had no killing effect on the CD4 + T cells of the blank control group. Conclusion Type 1 diabetes antisense peptide phage vaccine can induce CD8 + T cells to inhibit pathological CD4 + T cells.