HIV-1 gp41融合多肽对CD3抗体激活的调节性T细胞的抑制作用  

Inhibitory effects of HIV-1 gp41 fusion peptide on CD3 antibody activated regulatory T cells

作　　者：贺龙刚[1,2] 胡义平[1] 彭辉[3] 李珉珉[4] 刘叔文[1] 李晓娟[1] 

机构地区：[1]南方医科大学药学院,广东广州510515 [2]福建中医药大学药学院,福建福州350108 [3]中山大学医学院,广东广州510080 [4]暨南大学附属第一医院,广东广州510630

出　　处：《细胞与分子免疫学杂志》2012年第11期1121-1125,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(30801413;81073119);广东省医学科研基金(A2010327)

摘　　要：目的:观察HIV-1 gp41融合多肽(FP)能否影响CD3抗体激活的调节性T细胞(Treg)。方法:用免疫磁珠分离小鼠脾脏CD4+CD25+的Treg、CD4+CD25-的效应性T细胞(Teff),丝裂霉素C处理小鼠脾细胞获得APC。通过CCK-8法和羧基荧光素双乙酸盐琥珀酰亚胺酯(CFSE)标记检测FP对CD3抗体刺激的Teff增殖效应分析其对Treg抑制功能的影响;用ELISA检测FP对Treg激活后IL-10分泌的影响;用激光共聚焦显微镜观察细胞表面FP与TCR的分布。结果:通过CCK-8法和流式细胞术分析发现,在CD3抗体刺激下Teff细胞有显著增殖效应;当Treg和Teff混合培养时,Treg能够明显抑制共培养的Teff增殖;当加入25μg/mL FP后,Treg+Teff组与Teff组细胞增殖没有明显变化;当FP浓度为5μg/mL时,Treg+Teff组比Teff组增殖显著降低。Treg未活化时IL-10分泌较低,经过CD3抗体刺激后其IL-10分泌显著增多,而当FP浓度为25μg/mL时IL-10显著下降,5μg/mL FP对IL-10分泌无显著影响。通过激光共聚焦显微镜检测发现Treg未活化时,T细胞受体(TCR)在细胞表面均匀分布,同时未见FP与TCR共分布;当Treg在CD3抗体刺激下,TCR活化形成半月形,且FP与活化的TCR在细胞表面共分布。结论:25μg/mL FP对Treg体外抑制功能具有显著抑制作用,而5μg/mL FP不影响Treg的抑制功能,可能与其抑制Treg细胞IL-10分泌以及阻断APC在Treg细胞跨膜区域活化信号的递呈有关。AIM： To investigate whether the HIV-1 gp41 fusion peptide （FP） could affect the regulatory T cell （Treg） function activated by CD3 antibody. METHODS： Murine CD4＋ CD25 ＋Treg and CD4＋ CD25- effector T cells （Teff） were isolated from mice spleens by the immune magnetic beads. The splenocytes were treated with mitomycin C to obtain antigen presenting cells （APCs）. CCK-8 assay and CFSE loading were employed to evaluate the effects of FP on the Treg inhibitory function via measuring Teff prolifer- ation activated by CD3 antibody. In addition, IL-10 secretion of activated Treg was detected by ELISA. The distribution of FP and TCR on Treg surface was observed by laser scan- ning confocal microscope. RESULTS： Through the CCK-8 assay and flow cytometry, we found that Teff cells have significant proliferation stimulated by the CD3 antibodies. When Treg and Teff were co-cultured, the proliferation of Teff was significantly inhibited by Treg. When added with 25μg/mL FP, the proliferation of Treg ＋ Teff group and Teff group was not significantly affected, but when 5 μg/mL FP was added, the proliferation rate of Treg ＋ Teff group was significantly lower than that of Teff group. IL-10 secretion was low when Treg were not activated, but it significantly increased bv CD3 antibodies stimulation. When the concen-tration of FP was 25 μg/mL, IL-10 level significantly decreased, but 5 iag/mL FP did not significantly influence IL- 10 secretion. Through the laser scanning confocal micro- scope, we found that T cell receptors （TCR） of non-activa- ted Treg showed uniform distribution on the cell surface, and that FP and TCR had no common distribution. When Treg were stimulated by CD3 antibodies, the activated TCR formed half crescent, and FP and the activated TCR had common distribution on cell surface. CONCLUSION： Treg inhibitory function is significantly inhibited by 25 μg/mL FP in vitro, but 5 μg/mL FP does not affect it. This may be due to the suppression of IL-10 secretion and the influence o

关 键 词：HIV-1融合多肽 调节性T细胞 效应性T细胞 IL-10 

分 类 号：R293.1[医药卫生—民族医学]