SV40 Pathogenesis of Human Infections

描述（由申请人提供）：目前有说服力的证据表明，SV 40正在导致当今人群的感染。医学研究所最近得出结论，“生物学证据表明，在自然条件下接触SV 40可能导致人类癌症”，并建议“进一步研究SV 40在人类中的传播性”。与SV 40相关的特定人类肿瘤类型包括非霍奇金淋巴瘤。本提案的目的是评估SV 40在人类中的发病机制和传播性，并确定病毒大T抗原（T-ag）羧基（C）末端的可变结构域在病毒发病机制中是否具有重要功能。具体目标如下：（1）确定目标人群（器官移植受者）中SV 40感染的患病率、传播和发病率。我们将前瞻性评估成人移植患者外周血细胞和尿液中是否存在SV 40。将对SV 40毒株进行表征，并确定人体组织和恶性肿瘤中病毒基因组的状态和表达。将在SV 40感染和疾病之间建立临床和病理学相关性，包括系统性淋巴瘤的发展。(2)确定SV 40毒株变异对人外周血单核细胞（PBMC）中病毒复制和B细胞淋巴细胞永生化的影响。将检测不同SV 40毒株在PBMC亚群中的复制潜力和在B淋巴细胞中的永生化能力。将测定永生化B细胞中病毒DNA的状态和表达。将在感染的PBMC中监测病毒调控区的变化。(3)鉴定C-末端T-ag相互作用的细胞蛋白。将使用酵母双杂交系统，使用源自人类恶性肿瘤的SV 40 T-ag筛选人类淋巴细胞cDNA文库;将表征识别T-ag C末端结构域的相互作用蛋白;将比较与野生型和突变型T-ag的相互作用。该计划解决了我们对人类SV 40感染的传播和发病机制的知识中的重要空白。这将揭示SV 40毒株在人类中的致病作用是否不同，以及C-末端T-ag结构域在这些过程中的可能作用。这些新的信息将使我们更好地了解SV 40人类感染的生物学，并可能为SV 40参与人类恶性肿瘤的机制提供见解。

DESCRIPTION (provided by applicant): Persuasive evidence now indicates that SV40 is causing infections in the human population today. The Institute of Medicine recently concluded "the biological evidence is of moderate strength that SV40 exposure could lead to cancer in humans under natural conditions" and recommended "further study of the transmissibility of SV40 in humans". Specific human tumor types associated with SV40 include non-Hodgkin's lymphoma. The goals of this proposal are to evaluate the pathogenesis and transmissibility of SV40 in humans and to determine if the variable domain at the carboxy (C) terminus of the viral large T-antigen (T-ag), is functionally important in viral pathogenesis. The specific aims are the following: (1) Determine the prevalence, transmission, and morbidity of SV40 infections in a target human population (organ transplant recipients). We will evaluate prospectively the presence of SV40 in peripheral blood cells and in urine among adult transplant patients. SV40 strains will be characterized and the state and expression of the viral genome in human tissues and malignancies determined. Clinical and pathological correlations will be established between SV40 infections and disease, including the development of systemic lymphomas. (2) Determine the influence of SV40 strain variation on viral replication in human peripheral blood mononuclear cells (PBMCs) and on immortalization of B-cell lymphocytes. Different SV40 strains will be tested for replication potential in subsets of PBMCs and for immortalization ability in B lymphocytes. The state and expression of viral DNA in immortalized B-cells will be determined. Viral regulatory region changes will be monitored in infected PBMCs. (3) Identify C-terminal T-ag interactive cellular proteins. An SV40 T-ag derived from human malignancies will be used to screen a human lymphocyte cDNA library using the yeast two-hybrid system; interactive protein(s) that recognize the C-terminal domain of T-ag will be characterized; interactions with wild-type and mutant T-ags will be compared. This program addresses important gaps in our knowledge concerning the transmission and pathogenesis of SV40 infections in humans. It will reveal if SV40 strains vary in pathogenic effects in humans and the possible role of the C-terminal T-ag domain in those processes. This new information will allow better understanding of the biology of SV40 human infections and may provide insights into the mechanisms of SV40 involvement in human malignancies.