ANALYSIS AND ISOLATION OF SOLUBLE CD4 BINDING RETROVIRUS FROM THE PATIENTS WITH COLLAGEN DISEASES

本课题试图从外周血淋巴细胞和滑膜中分离出具有HIV相关逆转录病毒CD 4结合域的未知逆转录病毒。RA患者血清中与HIV gp 120结合的可溶性sCD_4浓度明显高于正常人。用逆转录聚合酶链反应（RT-PCR）方法，用扩增HIV gp 120非点突变区的引物，在类风湿关节炎（RA）患者外周血淋巴细胞和滑膜组织中，不能检测到这种未知的逆转录病毒mRNA。并应用荧光素差异显示技术检测高sCD 4 RA患者的疾病相关基因。本研究显示了外周血淋巴细胞的一些分化带。将其中一条单链DNA带亚克隆，并通过自动DNA测序仪分析DNA序列。DNA序列与人内源性逆转录病毒相似。本研究采用定量RT-PCR方法进一步研究了该基因在RA患者外周血淋巴细胞和滑膜中的表达情况，这可能是首次证实RA患者外周血淋巴细胞中存在未知的人内源性逆转录病毒，为进一步研究该内源性逆转录病毒与RA发病机制的关系奠定基础。

This project try to isolate unknown retrovirus which had CD4 binding domein of HIV related retrovirus in the peripheral blood lymphocytes and synovium. The concentration of to soluble (sCD4) which bound to gp120 in HIV,was significant higher in sera from patients with RA than normal individuals. This unknown retrovirus mRNA was not able to detect by reverse transcriptase polymerase chain reaction (RT-PCR) method using primers which amplified the non point mutated region of gp120 in HIV in the peripheral blood lymphocytes and sinovial tissue from the patients with rheumatoid arthritis (RA). Furthemore we employed Fluorescein Differencial Display method to detect the disease associated genes in the patients with RA with high concentration of sCD4 in sera. This study demonstrated the some differentiated bands in the periferal blood lymphocytes. One of these single stranded DNA bands was subcloned and analyzed the DNA sequence by the automatic DNA sequencer. The DNA sequence was similar to an human endogenous retrovirus. Now we are further studing the expression of this gnen in the peripheral lymphocytes and synovium from the patientes with RA by quantitative RT-PCR.This might be a first report that demonstrated the existence of unknown human endogenous retrovirus in periferal blood lymphocytes from the patients with RA.Further investigation will clarify the association between this endogenous retrovirus and pathogenesis of RA.