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目的探讨恶性疟原虫抗原对健康人外周血T淋巴细胞免疫功能的影响。方法健康成人外周血单个核细胞(PBMC)体外分别用恶性疟原虫(P.f)抗原(5μg/ml)和正常红细胞(nRBC)抗原(5μg/ml)进行刺激,同时给予IL-2维持细胞增殖,另外设立只加IL-2的阴性对照组进行培养。培养12 d时,刺激组细胞再用对应的抗原刺激20 h,用流式细胞仪检测T淋巴细胞亚群分泌IL-4和IFN-γ的情况,并用羧基荧光素乙酰乙酸琥珀酰亚胺酯(Carboxyfluorescein diacetate,succinimidyl ester,CFSE)标记法检测T细胞增殖反应。结果健康人PBMC经P.f抗原刺激扩增后CD4+T细胞的增殖指数(PI)明显高于nRBC抗原刺激组和阴性对照组(P均<0.05),但3组γδT的PI差异无统计学意义(P>0.05)。P.f抗原组分泌IL-4的CD4+T细胞百分率明显高于nRBC抗原组和阴性对照组(P均<0.05),但3组分泌IFN-γ的CD4+T细胞百分率差异无统计学意义(P>0.05)。结论 P.f抗原在体外可刺激健康人外周血CD4+T细胞增殖活化,后者通过优先分泌IL-4而发挥免疫调节作用。
Objective To investigate the effect of Plasmodium falciparum antigen on the immune function of T lymphocytes in peripheral blood of healthy volunteers. Methods Peripheral blood mononuclear cells (PBMCs) from healthy adults were stimulated with Pf antigen (5μg / ml) and normal erythrocyte antigen (5μg / ml) respectively in vitro. IL- In addition, a negative control group with IL-2 alone was set up. After cultured for 12 days, the stimulated cells were stimulated with the corresponding antigen for 20 h, the secretion of IL-4 and IFN-γ by T lymphocyte subsets was detected by flow cytometry, and the phosphorylation of carboxyfluorescein acetoacetate succinimidyl ester T cell proliferative response was detected by Carboxyfluorescein diacetate (succinimidyl ester, CFSE) labeling method. Results The proliferative index (PI) of CD4 + T cells in PBMCs stimulated with Pf antigen was significantly higher than that in nRBC antigen-stimulated group and negative control group (P <0.05), but the PI of γδT in 3 groups was not significantly different (P> 0.05). The percentage of CD4 + T cells secreting IL-4 in Pf antigen group was significantly higher than that in nRBC antigen group and negative control group (P <0.05), but the percentage of CD4 + T cells secreting IFN-γ in the three groups had no significant difference (P > 0.05). Conclusion P.f antigen can stimulate the proliferation and activation of CD4 + T cells in peripheral blood of healthy people in vitro, which play an immunomodulatory role by preferentially secreting IL-4.