日本血吸虫可溶性成虫抗原和虫卵抗原对CD4^+ T淋巴细胞凋亡和细胞周期的影响

Effect of soluble worm antigen and soluble egg antigen of Schistosoma japonicum on apoptosis and cell-cycle of CD4^+T lymphocytes

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 investigate and compare the different effects of soluble adult worm antigen （SWA） and soluble egg antigen （SEA） of Sehistosoma japonicum on the apoptosis and cell-cycle of murine CD4^ ＋ T cells. Methods Purified CD4^＋ T cells from normal C57BL/6 mice were cultured with CFSE labeled antigen presenting cells in the presence of different stimuli for 36 h. Flow cytometry （FCM） was used to detect the apoptosis of CD4^＋ T cells by fluorescence conjugated easpase-3 antibodie staining. The flow cytometry was used to analyze the cell-cycle of CD4^＋ T cells cultured as described above for 96 h by propidium iodide staining. Results Compared with the apoptosis percentage of CD4^＋ T cells [ （ 1.24 ± 0.29 ） % ] in the SEA stimulated group, that in the SWA stimulated group[ （ 1.52 ± 0. 38 ） % I did not show statistically significant difference （ P 〉 0.05 ）. Compared with the cell percentages in G1 phase [ （ 78.91 ± 2.98 ） % ], S phase [ （ 7.39 ±0. 85 ） % I and G2/M phase [ （ 10.69 ± 1.05） % I in the SWA stimulated group, that of the G1 phase [ （59.42 ± 1.32） % ] was significantly lower, but those in the S phase [ （21.07 ± 0.88） % I and G2/M phase[ （ 18.88 ± 1.21 ） % ] were significantly increased in the SEA stimulated group（ P 〈 0.01 ）. Conclusions There is no statistically significant difference between the apoptosis levels of CD4 ^＋ T cells stimulated by SWA and SEA. However, SEA significantly promotes the progression of the cell-cycle of CD4^＋ T cells compared with SWA.