多肽类结核杆菌抗原激活的γδ T细胞表达抗原提呈细胞表型和功能_临床医学论文.docVIP

多肽类结核杆菌抗原激活的γδ T细胞表达抗原提呈细胞表型和功能_临床医学论文 多肽类结核杆菌抗原激活的γδ T细胞表达抗原提呈细胞表型和功能_临床医学论文 【摘要】 目的: 观察用多肽类结核杆菌耐热抗原(Mtb激活的人外周血γδ T细胞是否具有抗原提呈细胞的表型和抗原提呈作用。方法: 外周血单个核细胞(PBMC)经贴壁法获取单核细胞, 加GM和LPS培养诱导为成熟DC。PBMC用Mtb优势扩增γδT细胞后用流式分选纯化γδ T(Vδ2+)细胞。PBMC经尼龙毛柱法获得纯化T细胞, 用CFSE标记后单独加Mtb分泌性抗原(Mtb或加单核细胞和Mtb共同培养, 或与经Mtb预处理的成熟DC或活化γδT细胞共同培养。第11天时用流式细胞术检测CD4+T细胞的增殖。Mtb活化的γδ T细胞与FITC标记的Mtb孵育不同时间后用流式细胞术和激光共聚焦显微镜检测摄入情况。结果: Mtb活化的γδ T细胞CD80, CD86和MHC类分子的表达显著增加。Mtb预处理活化的γδ T细胞诱导初始T 细胞增殖数和CD4+T细胞增殖指数的活性与成熟DC相似。Mtb活化的γδ T细胞能摄入FITC标记的Mtb。结论: 用多肽类 Mtb激活的γδ T细胞具有抗原提呈细胞的表型, 并具有摄入和提呈可溶性抗原给初始T淋巴细胞的能力。 【关键词】 结核分枝杆菌; 抗原提呈; T细胞亚群; 流式细胞术; 激光共聚焦显微镜 ［Abstract］ AIM: To explore whether the human peripheral γδ T cells stimulated by polypeptide heat resistant antigen from Mycobacterium tuberculosis (Mtblls by adding LPS. PBMCs were stimulated with MtbHAg and IL2 to expand predominantally γδ T cells that were further purified by flow sorting. The pure T cells were isolated from PBMCs using adhesion revomal and nylon column method, labeled with CFSE, and cultured in alone with Mtb secretory antigen (MtbSAg), in monocytes with MtbSAg, or with MtbSAg prepulsed dendritic cells or MtbHAg activated γδ T cells for 11 days. The proliferation of the CD4+T cells were measured by flowcytometry. The MtbHAg activated γδ T cells were incubated with FITC labelled MtbSAg for different time, and the ingestion of MtbSAg by γδ T cells was measured by flowcytometry and observed with laser confocal microscopye. RESULTS: The expressions of MHCII and costimulatory molecules CD80 and CD86 on γδ T cells that activated by MtbHAg marketly incrased in comparison with that on the resting γδ T cells. The expanded cell counts and proliferation index of the pure nave T cells that induced by MtbSAg pretreated activated γδ T cells were similar to that induced by mature DC. By using flowcytometer and laser confocal microscope, FITC labellaed MtbSAg was ingested by the activated γδ T cells. CONCLUSION: γδ T cells stimulated by polypeptide Mt