γ-氨基丁酸及其受体调控CD8^+T细胞活化  

Activation of CD8^+T cells regulated by γ-aminobutyric acid and its receptors

作　　者：樊文梅[1] 高钰[1] 孙玉洁[1] 马锡慧[1] 何秀云[1] 肖漓[1] 石炳毅[1] 

机构地区：[1]解放军第309医院全军器官移植研究所研究室,北京100091

出　　处：《器官移植》2017年第2期144-148,共5页Organ Transplantation

基　　金：国家自然科学基金(81270509)

摘　　要：目的探讨γ-氨基丁酸(GABA)及受体对CD8^+T细胞增殖能力的影响。方法采用CD8^+T细胞磁珠分选试剂盒分选Balb/c小鼠脾脏的CD8^+T细胞;在CD3/CD28活化磁珠的作用下,以不同浓度的GABA刺激CD8^+T细胞,通过5-溴脱氧尿苷(Brd U)标记和流式细胞术检测CD8^+T细胞的增殖情况;通过实时荧光定量聚合酶链反应(PCR)比较CD8^+T细胞活化前后GABA-A、GABA-B受体的表达。结果流式检测结果显示,GABA可抑制已活化的CD8^+T细胞的增殖,表现为CD152^+CD8^+T细胞明显减少;实时荧光定量PCR结果显示,仅在CD8^+T细胞活化时表达的GABA-A受体亚型为α2、α6和GABA-B受体亚型为1a;CD8^+T细胞活化后有明显增加的GABA-A受体亚型有α3、α5、β2、β3、γ1、γ2和θ,而GABA-B2R和GABA-B1b则在活化前和活化后均无表达。结论GABA可抑制已活化的CD8^+T细胞的增殖,其活化受GABA相关受体所调节,但其具体机制还有待于进一步探讨。Objective To evaluate the effect of γ-aminobutyric acid （GABA） and its receptors upon the proliferation of CD8＋T cells. Methods The splenic CD8＋T cells of Balb/c mice were obtained by CD8＋T cell magnetic bead sorting kit. Under the effect of CD3/CD28-activated magnetic bead, CD8＋T cells were stimulated by different concentrations of GABA. 5-bromo-2-deoxyuridine （BrdU） labeling and flow cytometry were performed to detect the proliferation of CD8＋T cells. The expression levels of GABA-A and GABA-B receptor before and after CD8＋T cell activation were compared by fluorescent quantitative real-time polymerase chain reaction （PCR）. Results Flow cytometry result revealed that GABA could inhibit the proliferation of activated CD8＋ cells, manifested as significant decrease in the quantity of CD152＋CD8＋T ceils. Fluorescent quantitative real-time PCR demonstrated that GABA-A receptor subtypes α2, αand GABA-B receptor subtype la were expressed only when the CD8＋T cells were activated. After CD8＋T cell activation, the quantity of GABA-A receptor subtypes α3, α5, β2, β3, γ1, γ2 and 0 were significantly increased, whereas the quantity of GABA-B2R and GABA-Blb did not significantly differ before and after CD8＋T cell activation. Conclusions GABA can suppress the proliferation of activated CD8＋T cells. The activation of CD8＋T cells is regulated by GABA receptors. However, the underlying mechanism remains to be elucidated.

关 键 词：CD8+T细胞 CD3/CD28 Γ-氨基丁酸 移植免疫 GABA-A受体 GABA-B受体 细胞增殖 T细胞活化 CD152 

分 类 号：R617[医药卫生—外科学] R392.4[医药卫生—临床医学]