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目的观察并比较日本血吸虫可溶性成虫抗原(SWA)、可溶性虫卵抗原(SEA)对小鼠CD4+T淋巴细胞凋亡和细胞周期的影响。方法以免疫磁珠分选正常小鼠脾细胞中的CD4+T淋巴细胞后,与羧基荧光素二醋酸盐琥珀酰亚胺酯标记的抗原递呈细胞共培养,并分别以PBS、SWA、SEA刺激。36h后以流式细胞术(FCM)结合caspase-3荧光抗体标记技术检测CD4+T淋巴细胞的凋亡水平;96h后以碘化丙锭染色结合FCM分析CD4+T淋巴细胞周期分布情况。结果凋亡分析结果显示,经SEA和SWA刺激后,外周CD4+T细胞的凋亡百分比分别为(1.24±0.29)%和(1.52±0.38)%,两者差异无统计学意义(P>0.05)。细胞周期分析表明,SWA组处于G1期、S期和G2/M期的细胞百分比分别为(78.91±2.98)%、(7.39±0.85%)和(10.69±1.05)%;与之相比,SEA组G1期细胞百分比[(59.42±1.32)%]显著降低,S期[(21.07±0.88)%]和G2/M期[(18.88±1.21)%]显著增加,差异均有统计学意义(P均<0.01)。结论SWA、SEA均可诱导CD4+T淋巴细胞凋亡,SEA促进CD4+T淋巴细胞进入分裂周期的作用更明显。
Objective To observe and compare the effects of soluble adult worm antigen (SWA) and soluble egg antigen (SEA) on the apoptosis and cell cycle of CD4 + T lymphocytes in mice. Methods CD4 + T lymphocytes were isolated from normal mouse splenocytes by immunomagnetic beads and co-cultured with carboxyfluorescein diacetate succinimidyl ester labeled antigen presenting cells. The cells were treated with PBS, SWA, SEA stimulation. After 36 hours, the apoptosis of CD4 + T lymphocytes was detected by flow cytometry (FCM) combined with caspase-3 fluorescent antibody labeling. The distribution of CD4 + T lymphocytes was analyzed by propidium iodide staining and FCM after 96 hours. Results The results of apoptosis analysis showed that the percentages of apoptotic CD4 + T cells in peripheral blood were (1.24 ± 0.29)% and (1.52 ± 0.38)% respectively after SEA and SWA stimulation, with no significant difference between the two groups (P> 0.05 ). Cell cycle analysis showed that the percentage of cells in the SWA group at G1, S and G2 / M phases was (78.91 ± 2.98)%, (7.39 ± 0.85%) and (10.69 ± 1.05)%, respectively The percentage of cells in group G1 was significantly decreased (59.42 ± 1.32% vs S.07.07 ± 0.88% vs G2 / M [18.88 ± 1.21%], P <0.05), and the difference was statistically significant (P All <0.01). Conclusion Both SWA and SEA can induce the apoptosis of CD4 + T lymphocytes and SEA can promote the entry of CD4 + T lymphocytes into the dividing cycle.