本文選題：L-選擇素 + γδT細(xì)胞　； 參考：《安徽醫(yī)科大學(xué)》2005年碩士論文

【摘要】：目的 (1)探討L-選擇素(CD62L)在結(jié)核桿菌抗原(Mtb-Ag)誘導(dǎo)人γδT細(xì)胞表達(dá)的動力學(xué)特征；(2)觀察不同細(xì)胞因子對活化人γδT細(xì)胞表達(dá)L-選擇素的調(diào)節(jié)作用；(3)應(yīng)用信號分子阻斷劑初步探討PKC，P13K，MAPK等信號途徑在人γδT細(xì)胞活化過程中對L-選擇素表達(dá)的調(diào)控作用。 方法 (1)正常人外周血單個核細(xì)胞分別用Mtb-Ag和CD3單抗(CD3mAb)刺激并加IL-2培養(yǎng)擴(kuò)增，分別獲得Mtb-Ag激活的T細(xì)胞(MtbAT)(γδT細(xì)胞為主)和CD3mAb激活的T細(xì)胞(CD3AT)(αβT細(xì)胞為主)。(2)取新鮮PBMC和刺激后培養(yǎng)3～21天的γδT細(xì)胞和αβT細(xì)胞，用多色熒光抗體標(biāo)記，在流式細(xì)胞儀上檢測γδT細(xì)胞表面L-選擇素的表達(dá)，并與αβT細(xì)胞作比較。(3)Mtb-Ag初次與再次刺激人γδT細(xì)胞分別培養(yǎng)7天，在培養(yǎng)同時分別按組加入IFN-γ、anti-IFN-γ、IL-12、anti-IL-12，并與培養(yǎng)的不同時間分別收集細(xì)胞，檢測L-選擇素表達(dá)情況。(4)培養(yǎng)6d～8d的MtbAT分別加入PMA、Ionomycin(IMN)、PMA+IMN培養(yǎng)3h、6h、12h、24h后，應(yīng)用流式細(xì)胞術(shù)(FCM)分別檢測L-選擇素的表達(dá)率。(5)培養(yǎng)8天的γδT細(xì)胞分別加入Mtb-Ag、Rotterin+Mtb-Ag、PMA、Rotterin+PMA刺激4h，，收集細(xì)胞并檢測L-選擇素表達(dá)。(6)MtbAT或CD3AT預(yù)先用LY294002(P13K途徑抑制劑)、SB203580(p38途徑抑制劑)、
[Abstract]:Objective 1) to investigate the kinetic characteristics of L- selectin CD62L) on the expression of human 緯 未 T cells induced by Mycobacterium tuberculosis antigen Mtb-Ag2) to observe the regulatory effect of different cytokines on the expression of L- selectin in activated human 緯 未 T cells.To explore the regulation of L- selectin expression by PKC- P13K- MAPK signaling pathway in the activation of human 緯 未 T cells.Methods 1) normal human peripheral blood mononuclear cells were stimulated with Mtb-Ag and CD3 monoclonal antibody CD3mAband cultured with IL-2.Mtb-Ag activated T cells (緯 未 T cells) and CD3mAb activated T cells (CD3ATA) were harvested from fresh PBMC and cultured for 321 days after stimulation. The cells were labeled with polychromatic fluorescent antibody.The expression of L- selectin. 4) the MtbAT of 6d~8d was added to PMA Ionomycinia IMN for 3 h, 6 h, 12 h and 24 h, respectively.The expression rate of L- selectin was detected by flow cytometry (FCM). 緯 未 T cells cultured for 8 days were added to Mtb-AgRotterin Mtb-AgPMA-Rotterin PMA for 4 h. The cells were collected and detected the expression of L- selectin. MtbAT or CD3AT were pretreated with LY294002(P13K pathway inhibitor SB203580p38.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R392

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