POLYOMAVIRUS IN HUMAN RENAL ALLOGRAFT TISSUE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 随着他克莫司和霉酚酸酯等免疫抑制药物的引入，一些主要医学中心的肾移植急性排斥反应的发生率已降至10%。不幸的是，这些强化治疗方案增加了病毒感染的风险。特别值得关注的是最近在大约5%的患者中出现了多瘤病毒异基因移植物肾病(PVAN)。PVAN是一种研究不足的令人痛苦的疾病，经常导致移植物丢失。由于多瘤病毒(PV)在高达50%的健康成年人的肾脏中潜伏，很可能较轻形式的PV相关移植物功能障碍得不到识别和治疗，导致持续的移植物损伤和慢性排斥反应的加速发展。这项建议的具体目的是(1)描述PV感染的完整临床谱，(2)描述影响人类PV复制的宿主和病毒因素(3)分析PV感染相关细胞基因在培养细胞、肾移植患者的血液和尿液以及泌尿生殖道癌组织标本中的表达，以及(4)进行抗BK病毒药物的体外测试，以便将来可能在临床领域使用。这些具体目标将通过以下方式实现：(A)在临床样本中进行多瘤病毒BK、JC和SV40的聚合酶链式反应，(B)进行核苷酸测序，以确定病毒基因型并确定病毒基因组中的基因突变和重排，(C)使用DNA微阵列技术、RT-PCR和ELISA来表征病毒诱导的宿主基因表达，以及(D)在流式细胞仪分析中测试可能的抗病毒药物的效果，该分析旨在测量培养细胞中的病毒复制率。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. With the introduction of immunosuppressive drugs such as tacrolimus and mycophenolate mofetil, the incidence of acute rejection in renal transplantation has fallen to <10% at some major medical centers. Unfortunately, these intensive therapeutic regimens increase the risk for vira infections. Of particular concern is the recent emergence of polyomavirus allograft nephropathy (PVAN) in approximately 5% of patients. PVAN is an inadequately studied and distressing condition that frequently leads to graft loss. Since polyomavirus( PV) is latent in the kidney of up to 50% of healthy adults, it is likely that milder forms of PV associated graft dysfunction go unrecognized & untreated, leading to ongoing graft damage and accelerated development of chronic rejection. The specific aims of this proposal are (1) T characterize the complete clinical spectrum of PV infection, (2) To delineate the host & viral factors that affect PV replication in man (3) To analyze PV infection associated cellular gene expression in cultured cells, blood and urine of kidney transplant patients, and tissue specimens with urogenital carcinoma, and (4) To perform in-vitro testing of anti-BK virus drugs for possible future use in the clinical arena. These specific aims will be accomplished by (a) Performing PCR for polyomaviruses BK, JC, and SV40 in clinical samples, (b) Nucleotide sequencing to assign viral genotypes and characterize genetic mutations and rearrangements in the viral genome, (c) Using DNA microarray technology, RT-PCR, & ELISA to characterize virus induced host gene expression, and (d) Testing the effect of putative anti-viral drugs in a flow cytometric assay designed to measure the rate of viral replication i cultured cells.