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[结核分枝杆菌肽抗原激活的人γδT细胞中抗原呈递细胞的表型表达及功能]

[Phenotype expression and function of antigen presenting cells in huaman gammadelta T cells activated by peptide antigen from Mycobacterium tuberculosis].

作者信息

Wang Jing, Li Bai-qing

机构信息

Department of Immunology, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 233030, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2009 Jul;25(7):588-91.

Abstract

AIM

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.

METHODS

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.

RESULTS

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.

CONCLUSION

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.

摘要

目的

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。

方法

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。

结果

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。

结论

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。

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