超抗原seb活化的nkt细胞亚群形态和分化途径的研究..docx

超抗原SEB活化的NKT细胞亚群形态和分化途径的 研究 [11-01-28 11:56:00 ] 作者：陈钰,郭业磊， 钟 编辑：studa20 【摘要】 目的：以NKT细胞形态和分化途径为焦点,研究超抗原SEB活化的 NKT细胞亚群的特征。方法：C57BL/J小鼠脾细胞分别经SEB或者ConA诱导, 收集体外扩增10d和5 d的淋巴细胞以及获取正常黏附性巨噬细胞。用荧光 抗体染色,流式细胞仪测定淋巴细胞和大淋巴细胞群表面 CD69的表达、NKT 细胞亚群百分数和不同NKT细胞亚群的分化途径。倒置显微镜 400倍下比较大 淋巴细胞的形态。结果：SEB活化的淋巴细胞和大淋巴细胞表面 CD69分子表达 由正常值的0.11%分别提高到55.00%和 68.95%。大细胞群中CD8+和 TCRVB8+NKT3胞亚群的百分数由原始的 0.36%和 0.81%分别提高到30.29%和 31.48%。这些细胞位于流式细胞图的上部，是大体积细胞群。显微镜下显示 SEB活化的大淋巴细胞体积不仅大于 ConA活化的T淋巴细胞,而且比正常巨噬 细胞大5倍以上，是非黏附性细胞。胞内质粒松散、 胞质与核之比〉1。SEB 活化的CD8+和TCR/ 8+NKT细胞亚群均由T细胞直接分化而来。结论：SEB活 化 10 d 的 NKT细胞亚群主要是 CD8+ NK1.1+和 TCR/ 8+ NK1.1 +的 NKT细胞。 这些细胞是大体积淋巴细胞，应属于T细胞的亚群。 【关键词】 NKT细胞亚群；超抗原；细胞形态；细胞分化 [Abstract ] AIM: This study is to explore the biological characteristics of NKT subsets through inv estigati ng their morphology and differe ntiati on pathways. METHODS: Sple nic lymphocytes from C57BL/J mice were treated with either Staphylococcal en tertox in B (SEB) or ConA and then cultured in vitro. On day 5 and day 10, the lymphocytes were harvested. Normal adhere nt macrophages and no rmal lymphocytes were harvested on day 0. Expressi on of CD69 molecule on the cell surfaces of the lymphocytes and large lymphocytes by SEB activated was measured using sta ining of fluoresce nt an tibody and flow cytometry (FACS). And the perce nt (%) of NKT subsets was determ ined and an alyzed the differe ntiati on pathways of the NKT subsets. Appeara nee of subsets was observed and compared un der a light microscope at x 400 magn ificati on. RESULTS: Expressi on levels of CD69 molecule on the SEB activated lymphocytes and the gia nt lymphocytes were sig nifica ntly in creased to 55% and 68.95% respectively as compared to the level (0.11%) detected on the no rmal lymphocytes (both P0.01; n=3). The perce nt (%) of CD8+NK1.1+ and TcRVB 8+NK1.1+ NKT subsets in the giant lymphocytes was significantly enhanced to 30.29% and 31.48% (84.0 or 38.9 folds) origi nally from 0.36% and 0.81%, respectively. Based on the cell distributi on show n