PvMSP1-19重组蛋白体外对间日疟患者外周血T细胞免疫功能的影响

目的 探讨PvMSP1-19重组蛋白对间日疟既往感染者外周血T淋巴细胞功能的影响。方法 间日疟既往感染者外周血单个核细胞(PBMC),在体外分别以PvMSP1-19重组蛋白和IL-2(刺激组)及IL-2(未刺激组)培养,培养第7d时刺激组细胞再用PvMSP1-19重组蛋白刺激24h,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;并以CFSE标记法检测T淋巴细胞的增殖反应。结果 在分泌IL-4的淋巴细胞中,CD8⁺T细胞所占的比例刺激组(8.04%)明显高于未刺激组(3.46%); 而分泌IFN-(的CD8⁺T细胞在刺激组(0.82%)与未刺激组(1.05%)间并无显著性差别。分泌IL-4和IFN-(的CD4⁺T细胞比例在刺激组(1.81%,0.19%)与未刺激组(1.89%,0.05%)间并无显著性差异。此外,CD8⁺T细胞的增殖指数刺激组(5.65%)也明显高于未刺激组(3.69%)。结论 PvMSP1-19重组蛋白能显著诱导间日疟既往感染者外周血CD8⁺T细胞增殖和优先分泌IL-4。

Immune effect of T lymphocytes from Plasmodium vivax-infected patients stimulated by recombinant PvMSP1-19 antigen in vitro

In order to study the effect of T cells from Plasmodium vivax (P.vivax)-infected patients which were stimulated with PvMSP1-19 in vitro, the peripheral blood mononuclear cells (PBMCs) isolated from P. vivax-infected patients was stimulated with PvMSP1-19 protein, and cultured in rIL-2 containing media to generate PvMSP1-19 activated T cells. After 7 days of culture, PvMSP1-19 was used to restimulate the activated T cells for 24 hours. Then the cells were collected for detecting intracellular cytokines by Flow cytometry (FCM), and the cells-proliferations of T cells labeled by CSFE was also analysed by FCM. The percentages for IL-4 ⁺ T cells among activated CD8⁺ T cells that were stimulated with PvMSP1-19 was distinctly higher than that in the unrestimulated control group (P<0.05). Meanwhile, PI (proliferative index) of CD8⁺ T cells that were stimulated with PvMSP1-19 was distinctly higher than that in the unrestimulated control group (P<0.05). In conclusion, the recombinant PvMSP1-19 protein could effectively stimulate the proliferation of CD8⁺ T cells and the production of IL-4 secreted by CD8⁺ T cells.