Wang Jing, Li Bai-qing
Department of Immunology, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 233030, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2009 Jul;25(7):588-91.
To explore whether the human peripheral gammadelta T cells stimulated by polypeptide heat resistant antigen from Mycobacterium tuberculosis (Mtb-HAg) express phenotype and have the fuction of antigen presentation.
The monocytes isolated by adhesion method from peripheral blood mononuclear cells (PBMCs) were cultured with GM-CSF, IL-4 and induced into the mature dendritic cells by adding LPS. PBMCs were stimulated with Mtb-HAg and IL-2 to expand predominantally gammadelta 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 gammadelta T cells for 11 days. The proliferation of the CD4(+) T cells were measured by flowcytometry. The Mtb-HAg activated gammadelta T cells were incubated with FITC labelled Mtb-SAg for different time, and the ingestion of Mtb-SAg by gammadelta T cells was measured by flowcytometry and observed with laser confocal microscopye.
The expressions of MHC-II and co-stimulatory molecules CD80 and CD86 on gammadelta T cells that activated by Mtb-HAg marketly incrased in comparison with that on the resting gammadelta T cells. The expanded cell counts and proliferation index of the pure naïve T cells that induced by Mtb-SAg pretreated activated gammadelta 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 gammadelta T cells.
gammadelta T cells stimulated by polypeptide Mtb-HAg express the phenotype of professional antigen-presenting cells and they are able to present Mtb-SAg to naïve T cells to induce the proliferation of CD4(+) T cells.
探讨结核分枝杆菌耐热多肽抗原(Mtb-HAg)刺激人外周血γδT细胞是否表达表型并具有抗原呈递功能。
采用黏附法从外周血单个核细胞(PBMCs)中分离单核细胞,用GM-CSF、IL-4培养,并加入LPS诱导其成为成熟树突状细胞。用Mtb-HAg和IL-2刺激PBMCs以扩增主要为γδT细胞,再通过流式分选进一步纯化。用去除黏附及尼龙柱法从PBMCs中分离纯化T细胞,用CFSE标记,分别与Mtb分泌抗原(Mtb-SAg)单独培养、与含Mtb-SAg的单核细胞共同培养、与预先用Mtb-SAg脉冲处理的树突状细胞或Mtb-HAg激活的γδT细胞共同培养11天。通过流式细胞术检测CD4(+)T细胞的增殖情况。将Mtb-HAg激活的γδT细胞与FITC标记的Mtb-SAg孵育不同时间,通过流式细胞术检测γδT细胞对Mtb-SAg的摄取情况,并通过激光共聚焦显微镜观察。
与静息γδT细胞相比,Mtb-HAg激活的γδT细胞上MHC-II及共刺激分子CD80和CD86的表达明显增加。Mtb-SAg预处理激活的γδT细胞诱导的纯幼稚T细胞的扩增细胞数和增殖指数与成熟树突状细胞诱导的相似。通过流式细胞仪和激光共聚焦显微镜观察,激活的γδT细胞摄取了FITC标记的Mtb-SAg。
多肽Mtb-HAg刺激的γδT细胞表达专业抗原呈递细胞的表型,且能够将Mtb-SAg呈递给幼稚T细胞以诱导CD4(+)T细胞增殖。