MOLECULAR ANALYSIS OF HUMAN NAIVE AND MEMORY T CELLS

总结：本项目的目标是了解淋巴细胞分化的分子基础，从na？ve to memory记忆，and immunological免疫memory记忆.作为实现这一目标的第一步，我们分析了人类幼稚和记忆CD 4 + T细胞的基因表达谱，并确定了在一个群体中差异表达的基因。人CD 4+初始T细胞和记忆T细胞是表型上明确定义的亚群，即表达CD 45 RA的初始CD 4 + T细胞和表达CD 45 RO的记忆CD 4 + T细胞。我们采用免疫磁珠分离法从外周血中分离出大量高纯度的幼稚细胞和记忆细胞。从初始和记忆T细胞中分离信使RNA并用于基因表达分析。基因表达谱芯片分析基因表达谱及差异表达基因。大约一半的估计总人类基因（45,000）进行了分析，发现na？ve和记忆性CD 4 + T细胞表达的基因约占总基因的15-20%。在幼稚细胞和记忆细胞之间，约85%的表达基因的表达水平相似。使用的表达率为2作为标准的差异表达，我们发现，约300个基因（总表达基因的4%），是差异表达的na？ve CD 4 + T细胞和记忆性CD 4 + T细胞中约740个差异表达基因（占总表达基因的9%）。在这些基因中，只有5-10%的差异表达的已知基因，其中大多数是EST。目前，我们正在通过北方，原位杂交和测序来鉴定这些差异表达的基因。- 初始和记忆T细胞，基因芯片，淋巴细胞活化，无能

Summary: The goal of this project is to understand the molecular basis of lymphocyte differentiation from na?ve to memory, and immunological memory. As the first step toward this goal, we have analyzed gene expression profiles of human naive and memory CD4+ T cells, and identified genes that are differentially expressed in one population over the other. Human CD4+ naive and memory T cells are phenotypically well defined subsets, i.e. naive CD4+ T cells expressing CD45RA and memory CD4+ T cells expressing CD45R0. We used immunomagnetic separation isolating large number of highly pure naive and memory cells from peripheral blood. Messenger RNA was isolated from naive and memory T cells and used for gene expression analysis. Gene expression profile and differentially expressed genes were analyzed by cDNA microarray. Approximately half of the estimated total human genes (45,000) were analyzed and found that na?ve and memory CD4+ T cells expressed about 15-20% of total genes. The levels of expression about 85% of the expressed genes are similar between naive and memory cells. Using an expression ratio of 2 as criteria of differential expression, we found that about 300 genes (4% of total expressed genes) that are differentially expressed in na?ve CD4+ T cells and about 740 genes (9% of total expressed genes) differentially expressed in memory CD4+ T cells. Among these genes, only 5-10% of these differentially expressed known genes and the majority of them are ESTs. Currently, we are characterizing these differentially expressed genes by Northern, in situ hybridization, and sequencing. - Naive and memory T cells, cDNA microarray, lymphocyte activation, anergy