Gene expression analysis in naïve and vaccine (peptide vs. DC) primed pmel-1 CD8 Tcells. Mus musculus

We have shown that DC vaccine is superior to peptide vaccine in terms of priming and expansion of antigen-specific CD8+ T cells. DC vaccine-primed pmel-1 cells displayed better effecter functions than cells by peptide-primed cells in terms of cytokine production and externalization of cytotoxic granules. Furthermore DC vaccine-primed cells were metabolically distinct from peptide-primed cells. To confirm these findings, we performed a microarray analysis using splenic pmel-1 T cells from mice immunized with hgp100 peptide vaccine or DC vaccine. We also used splenic naïve pmel-1 T cells as a control. Overall design: C57BL/6 mice transferred with pmel-1 splenocytes were treated with hgp100 peptide vaccine or DC vaccine twice on days 0 and 14. On day 28, pmel-1 CD8+ T cells were sorted from harvested splenocytes of immunized mice. Gene expression was measured in naïve pmel-1 CD8+ T cells and sorted pmel-1 CD8+ T cells

本研究证实，在抗原特异性CD8+ T细胞的致敏与扩增方面，DC疫苗（DC vaccine）优于肽疫苗（peptide vaccine）。经DC疫苗致敏的pmel-1细胞，在细胞因子产生与细胞毒性颗粒外化方面，其效应功能优于肽疫苗致敏的pmel-1细胞。此外，经DC疫苗致敏的细胞在代谢特征上与肽疫苗致敏的细胞存在显著差异。为验证上述发现，本研究采用经hgp100肽疫苗（hgp100 peptide vaccine）或DC疫苗免疫的小鼠脾脏来源pmel-1 T细胞进行基因芯片（microarray）分析，并以初始脾脏pmel-1 T细胞作为对照。整体实验设计：将过继转移pmel-1脾细胞的C57BL/6小鼠分别于第0天和第14天两次接受hgp100肽疫苗或DC疫苗免疫。于第28天，从免疫小鼠的收获脾细胞中分选pmel-1 CD8+ T细胞。分别对初始pmel-1 CD8+ T细胞及分选得到的pmel-1 CD8+ T细胞进行基因表达量检测。