自噬小体疫苗能够检测人外周血抗原特异性T细胞-东南大学学报医学版

自噬小体疫苗能够检测人外周血抗原特异性T细胞

单位：1. 东南大学医学院, 江苏南京 210009;
2. 东南大学附属南京市第二医院生物治疗中心, 江苏南京 210003

分类号：R730.3

出版年·卷·期（页码）：2014·33·第四期（410-415）

摘要：

目的：探讨自噬小体疫苗DRibbles是否可用于检测人外周血中抗原特异性T细胞。方法：制备表达CEF蛋白的HEK293T细胞及表达CMV pp65蛋白的LT3细胞，从中提取DRibbles，用HEK293T CEF DRibbles及LT3 pp65 DRibbles刺激人外周血单个核细胞（PBMCs），通过流式细胞术检测分泌IFN-γ的T细胞占总T细胞的百分比。结果：与阴性对照HEK293T GFP DRibbles相比，HEK293T CEF DRibbles刺激组分泌IFN-γ⁺的CD8⁺ T细胞比例显著增高（3.38% vs 0.05%），分泌IFN-γ⁺的CD4⁺T细胞比例亦显著增高（0.46% vs 0.06%）；LT3 pp65 DRibbles刺激组分泌IFN-γ⁺的CD8⁺T细胞平均百分数为0.21%，阴性对照LT3 GFP DRibbles刺激组仅为0.05%（P<0.01）；与LT3pp65细胞裂解产物相比，LT3 pp65 DRibbles能诱导更多的CD8⁺T细胞分泌IFN-γ（P<0.05），在诱导CD4⁺ T细胞分泌IFN-γ方面，DRibbles与细胞裂解产物类似（P>0.05）。结论：包含病毒抗原的DRibbles能够有效激活人PBMCs中的病毒抗原特异性CD8⁺ T细胞及CD4⁺T细胞。

Objective: To explore if autophagosome vaccine DRibbles can be used to detect the antigen-specific T cell response in human peripheral blood. Methods: The DRibbles were extracted from HEK293T cells expressing CEF protein and LT3 cells expressing CMV pp65 protein. Then HEK293T CEF DRibbles and LT3 pp65 DRibbles were used to stimulate human PBMCs. The percentages of T cells secreting IFN-γ were detected by Flow cytometry intracellular staining. Results: Compared with the negative control HEK293T GFP DRibbles group, the percentage of IFN-γ⁺CD8⁺ T cells was increased dramatically(3.38% vs 0.05%) in HEK293T CEF DRibbles group.And the percentage of IFN-γ⁺CD4⁺ T cells was increased too(0.46% vs 0.06%). The mean of the percentage of IFN-γ⁺CD8⁺T cells was 0.21% in LT3 pp65 DRibbles group and only 0.05% in the LT3 GFP DRibbles group(P<0.01). Compared with the LT3 pp65 lysate LT3 pp65 DRibbles could induce more CD8⁺ T cells (P<0.05) and similar CD4⁺ T cells to secret IFN-γ(P>0.05). Conclusion: The DRibbles containing virus antigen can efficiently activate the virus antigen specific CD8⁺ and CD4⁺ T cells in human PBMCs.

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