摘要
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 pathologic CD4^+ T cells induced by diabetes 1 antisense peptide phage vaccine. Methods Non-obese diabetic (NOD) mice were divided into test, parallel control and blank control groups. The mice in test and parallel control groups were immunized twice with diabetes 1 antisense peptide phage vaccine and empty phage carrier respectively, while those in blank control group were unimmunized. On week 20 after the first immunization, the mice in various groups were determined for blood sugar contents, and their CD8^+ T cells were isolated with immunomagnetie beads, then stimulated with synthetic antisense peptide and IL-2 and used as effecter cells. The CD4^+ T cells of mice in parallel and blank control groups were isolated, then stimulated with synthetic positive-sense peptide and IL-2 and used as target cells. The effecter and target cells were mixed at various ratios and determined for CTL killing activity by lactate dehydrogenase (LDH) release test. Results On week 20 after the first immunization, the blood sugar of mice in test group were at a normal level, while those in parallel and blank control groups were higher than normal level. The killing rate of CD8^+ T cells of mice in test group to CD4^+ T cells of mice in parallel control group reached a peak value of 47. 95% ± 11. 30% at an effecter-target ratio of 100:1. However, the CD8^+ T cells of mice in parallel control group showed no killing activity to the CD4^+ T cells of mice in blank control group. Conclusion Diabetes 1 antisense peptide phage vaccine induced the inhibitory effect of CD8^+ T cells on pathologic CD4^+ T cells.
出处
《中国生物制品学杂志》
CAS
CSCD
2009年第8期779-781,共3页
Chinese Journal of Biologicals
基金
国家863项目"预防性1型糖尿病反义肽抗原的中试和临床前研究"(2006AA02Z4C0)
云南省科技厅攻关项目"噬菌体展示新型疫苗技术平台的建立及其在1型糖尿病反义肽疫苗研制中的应用"(2006SG05)
云南省自然科学基金面上项目"反义肽免疫诱导调节性T细胞的机理研究"(2006C0061M)