T淋巴细胞活化诱导的细胞死亡在肾综合征出血热发病中的作用  

The pathogenetic role of peripheral T-lymphocytes activation-induced cell death in hemorrhagic fever with renal syndrome

作　　者：王晓燕[1] 康鹏[1] 高杰[1] 于建武[1] 吕淑兰[1] 

机构地区：[1]哈尔滨医科大学附属第二医院感染病科,150086

出　　处：《中华内科杂志》2006年第11期887-890,共4页Chinese Journal of Internal Medicine

基　　金：黑龙江省自然科学基金资助项目(D01-48)

摘　　要：目的研究肾综合征出血热(HFRS)患者外周血T淋巴细胞(PBL-T)活化诱导细胞死亡(activation-inducedcelldeath,AICD)的发生情况,探讨AICD在HFRS发病机制中的作用。方法以免疫磁珠负性分离PBL-T;应用植物血凝素(PHA)和离子霉素(ionomycin)体外培养诱导细胞活化;采用HE染色和脱氧核苷酸原位末端标记法(TUNEL)染色观察凋亡情况并用流式细胞仪检测PBL-T的凋亡率。应用Westernblot检测HFRS患者外周血T淋巴细胞caspase-3的表达情况,用RT-PCR对汉坦病毒的特异性基因片段进行检测。结果HFRS患者诱导组PBL-T凋亡率为(27.79±0.99)%,明显高于未诱导组的(17.16±1.14)%(P<0.01);无论是否诱导,患者PBL-T的凋亡率均高于健康人。HFRS患者诱导组PBL-Tcaspase-3的17000活性成分表达显著增加。扩增产物测序结果,有19例患者血清出现阳性条带,其中汉滩病毒7例,汉城病毒12例。结论以PHA和离子霉素可有效地诱导体外培养的PBL-T发生AICD。HFRS患者PBL-T经激活诱导后凋亡率明显提高。有79.2%的患者可以扩增到汉坦病毒特异性的基因片段。HFRS患者PBL-T凋亡相关蛋白caspase-3的表达增加,提示AICD可能与HFRS的发病机制有关。Objective To study the activation-induced cell death （AICD） in peripheral blood T- lymphocytes（PBL-T） from patients of hemorrhagic fever with renal syndrome（HFRS） and the effect of AICD on the pathogenesis of HFRS. Methods The PBL-T of patients with HFRS were isolated by negative selection with magnetic beads and cultured with or without phytohemagglutinin （PHA） and ionomycin. The apoptotic ratio of PBL-T was detected with TUNEL staining and assessed with flow cytometry. The level of caspase-3 of PBL-T in patients with HFRS was detected with Western blot. Characteristic gene fragment of Hantaan virus and Seoul virus was detected with reverse transcription polymerase chain reaction. Results In HFRS patients, the apoptotic ratio （27. 79 ± 0. 99） % of PBL-T activated with PHA and ionomycin was significantly higher than that without activation （ 17. 16 ± 1. 14 ） %, （ P 〈 0. 01 ）. The apoptotic ratio of PBL- T in HFRS patients was higher than that of healthy control（ P 〈 0. 01 ）, no matter with or without activation. The expression level of caspase-3 increased in the induced group of patients with HFRS and the expression level of 17 000 （ the active component of caspase-3 ） increased significantly. The results of sequencing showed that amplification production from the serum are positive in 19 cases, seven being Hantaan virus and twelve Seoul virus. Conclusions AICD in PBL-T was found in patients of with HFRS. The high expression rate of caspase-3 in the PBL-T of patients with HFRS was related with AICD in the patients with HFRS. In the HFRS patients with specific IgM antibody positive, characteristic fragments can be obtained with RT-PCR from 79.2% patients. AICD of T lymphocytes may play an important role in the pathogenesis of HFRS.

关 键 词：肾综合征出血热 T淋巴细胞 脱噬作用 CASPASE-3 

分 类 号：R512.8[医药卫生—内科学]