恒河猴TRIM5α的组织分布及不同刺激促进PBMC中TRIM5α转录水平的上调  

Tissue Distribution of TRIM5α in Rhesus Monkey and Upregulation in Peripheral Blood Mononuclear Cell by Using Different Stimuli

作　　者：汤霞[1,2] 戴正喜[1,2] 夏厚军[1,2] 刘丰亮[1,2] 况轶群[1,2] 张高红[1] 郑永唐[1] 

机构地区：[1]中国科学院昆明动物研究所中国科学院和云南省动物模型与人类疾病机理重点实验室,云南昆明650223 [2]中国科学院研究生院,北京100049

出　　处：《Zoological Research》2009年第4期354-360,共7页动物学研究（英文）

基　　金：国家重点基础研究发展计划(973) (2006CB504302);国家自然科学基金(30671960;U0832601;30872317;30800113);中国科学院知识创新工程重要方向(KSCX1-YW-R-15);云南省科技基础条件平台计划(2006PT08)资助课题

摘　　要：TRIM5α(tripartite motif protein 5-alpha)蛋白是恒河猴体内一种非常重要的限制因子,能抑制人免疫缺陷病毒(HIV-1, human immunodeficiency virus type 1)、马感染性贫血病毒(EIAV, equine infectious anemia virus)和猫免疫缺陷病毒 (FIV, feline immunodeficiencyvirus)等逆转录病毒的复制。恒河猴 TRIM5α的组织分布以及在受到外界刺激时 TRIM5α mRNA 表达量的变化研究还未见报道。本研究从中国恒河猴的各组织中提取总 RNA,以β-actin基因作为内参照,通过半定量 RT-PCR 检测各组织中 TRIM5α mRNA 的表达。选择 HIV-GFP-VSVG 假病毒感染外周血单核细胞(peripheral blood mononuclear cell,PBMC),非特异性刺激剂——佛波脂(Phorbol myfismte acetate,PMA)+离子霉素(ionomycin,Ion)及 CD28 抗体+CD49d 抗体分别共刺激恒河猴 PBMC,研究不同刺激对恒河猴TRIM5α mRNA 表达水平的影响。结果表明:TRIM5α mRNA 表达于所研究的恒河猴 21 种组织中,免疫系统和泌尿生殖系统组织中表达量最高,而神经系统组织,如大脑、脊髓中表达较少,其他组织中未见明显的表达差异;HIV-GFP-VSVG 感染和用 PMA+Ion、CD28 抗体+CD49d 抗体分别共刺激 PBMC 能促进 PBMC 中 TRIM5α mRNA的转录水平的上调。TRIM5α （tripartite motif protein 5-alpha） protein is a very important restriction factor in rhesus monkey. It can restrict the replication of retroviruses, such as HIV-1 （human immunodeficiency virus type 1）, EIAV （equine infectious anemia virus） and FIV （feline immunodeficiency virus）. Up to now, there is no report about the tissue distribution of TRIM5α in rhesus monkey. The influence of different stimuli on the expression of TRIM5α mRNA has also not been examined in monkey peripheral blood mononuclear cell （PBMC）. In this study, total RNA was extracted from different tissues of a rhesus monkey collected in Yunnan, China, and was quantified using the β-actin gene as the internal reference. The expression levels of TRIM5α mRNA in PBMC that was treated with three conditions （the HIV-GFP-VSVG pseudotyped virus challenged, PMA and Ion-costimulated, anti-CD28 and anti-CD49d antibody-costimulated） were also evaluated at different time points. Our results showed that TRIM5α mRNA expressed broadly in all 21 studied tissues, with the highest level in tissues of immune system and urogenital system and a lower level in nervous system, such as cerebrum and spinal cord. The expression level of the TRIM5α mRNA level in PBMC was upregulated after being activated by the three treatments.

关 键 词：TRIM5α 恒河猴 组织分布 PBMC 共刺激 表达水平 

分 类 号：Q78[生物学—分子生物学]