Polyomavirus Infection after Kidney Transplantation

描述（由申请人提供）： 随着他克莫司和霉酚酸酯等免疫抑制药物的引入，在一些主要的医疗中心，肾移植中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，最近在约5%的患者中出现了多瘤病毒同种异体移植肾病（PVAN）。PVAN是一种研究不足且令人痛苦的疾病，经常导致移植物丢失。由于多瘤病毒（PV）潜伏在高达50%的健康成人的肾脏中，因此可能无法识别和治疗与PV相关的移植物功能障碍的轻度形式，导致持续的移植物损伤和慢性排斥反应的加速发展。本提案的具体目的是：（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）分析培养细胞、肾移植患者血液和尿液以及泌尿生殖道癌组织标本中PV感染相关的细胞基因表达;及（4）进行抗BK病毒药物的体外测试，以供将来可能用于临床竞技场。这些特定目的将通过以下方式实现：（a）对临床样本中的多瘤病毒BK、JC和SV 40进行PCR，（B）进行核苷酸测序，以分配病毒基因型并表征病毒基因组中的遗传突变和重排，（c）使用DNA微阵列技术、RT-PCR和ELISA表征病毒诱导的宿主基因表达，和（d）在设计用于测量培养细胞中病毒复制速率的流式细胞术测定中测试推定的抗病毒药物的效果。

DESCRIPTION (provided by applicant): 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 viral 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 kidneys of up to 50% of healthy adults, it is likely that milder forms of PV associated graft dysfunction go unrecognized and untreated, leading to ongoing graft damage and accelerated development of chronic rejection. The specific aims of this proposal are (1) To characterize the complete clinical spectrum of PV infection; (2) To delineate the host and 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, and 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 in cultured cells.