Developing a Gene Expression Signature For T Cells Resident in Human Skin. Homo sapiens

Human skin contains various populations of memory T cells in permanent residence and in transit. The most well characterized subset of resident memory T cell is the CD8+ CD103+ T cell. In order to investigate the remaining subsets, we isolated skin-tropic helper T cells, T regulatory cells, and CD8+ CD103- T cells from skin and from blood by fluorescence activated cell sorting. We extracted RNA and performed a microarray analysis to compare the transcriptional profile of these groups. We found that despite expressing the same marker for skin tropism, the T cells of skin origin were significantly different from those from blood. Overall design: Total RNA obtained from CLA+ Tregulatory cells, CLA+ helper T cells, and CLA+ CD8+ CD103- T cells isolated from human skin compared to CLA+ Tregulatory cells, CLA+ memory helper T cells, and CLA+ memory CD8+ T cells from human blood. 5 replicates of each; each replicate contains pooled cells from 3 subjects; total 15 subjects.

人体皮肤中存在多种常驻及循环中的记忆性T细胞亚群。其中研究最为透彻的常驻记忆性T细胞亚群为CD8+ CD103+ T细胞。为探究其余未被充分表征的亚群，本研究通过荧光激活细胞分选术（fluorescence activated cell sorting, FACS）从人体皮肤及外周血中分离出皮肤归巢辅助性T细胞、调节性T细胞（T regulatory cells, Treg）以及CD8+ CD103- T细胞。我们提取各组细胞的总RNA并开展基因芯片分析，以比较不同组间的转录组特征。研究发现，尽管这些T细胞均表达相同的皮肤归巢标志物，但皮肤来源的T细胞与血液来源的T细胞之间存在显著差异。
实验设计：本研究将从人体皮肤中分离得到的皮肤淋巴细胞相关抗原（cutaneous lymphocyte-associated antigen, CLA）阳性调节性T细胞、CLA阳性辅助性T细胞及CLA阳性CD8+ CD103- T细胞的总RNA，与从人体外周血中分离得到的CLA阳性调节性T细胞、CLA阳性记忆性辅助性T细胞及CLA阳性记忆性CD8+ T细胞的总RNA进行对比分析。每组设置5个生物学重复，每个重复混合3名受试者的细胞，共计纳入15名受试者。