Modeling KSHV Infection and Pathogenesis in SCID Mice

SCID 小鼠 KSHV 感染和发病机制建模

• 批准号: 7007044
• 负责人: GEROLD FEUER
• 金额: $ 19.61万
• 依托单位: UPSTATE MEDICAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7007044
• 关键词: B cell lymphoma; B lymphocyte; CD34 molecule; Herpesviridae disease; SCID mouse; antineoplastics; antiviral agents; apoptosis; carcinogenesis; cell differentiation; combination chemotherapy; disease /disorder model; hematopoiesis; hematopoietic stem cells; human herpesvirus 8; human tissue; interferon alpha; latent virus infection; model design /development; recombinant virus; stem cell transplantation; tissue /cell culture; virus infection mechanism; zidovudine

DESCRIPTION (provided by applicant): In vivo models are critical to the understanding of pathogenic mechanisms operating in viral diseases, and for evaluation of therapeutic approaches to combat these diseases. KSHV has been isolated from B cell derived lymphomas found in patients with AIDS, termed PEL and MCD. There is an incomplete understanding of the initial cellular targets of KSHV infection and cells that contribute to viral dissemination and latency in vivo. We speculate that KSHV infection of progenitor B cells is key process in the initiation of oncogenesis. KSHV genomic sequences have been detected in hematopoietic progenitor (CD34+) cells from KS patients. We have demonstrated that KSHV infects human CD34+ cells and that viral gene expression is maintained in CFU-GM clonogenic colonies derived from the differentiation of these cells in vitro. KSHV genomic DNA and LANA-1 expression was detected in human (CD14+) monocytes and in B lymphocytes from NOD/SCID-hu chimeric mice following reconstitution of lymphopoiesis with infected human CD34+ cells. Although this model system is not perfectly representative of normal human hematopoiesis and/or all aspects of KSHV infection, the combination of KSHV persistence in NOD/SCID-hu mice and infection of CD34+ cells provides an experimental system in which to address many of the issues of KSHV replication, latency and pathobiology in vivo. This novel animal model will be employed to assay the efficacy of AZT and IFN-alpha in suppressing KSHV infection in vivo. This proposal aims to further refine and develop the NOD/SCID-hu mouse model of KSHV infection and to better understand the role of KSHV infection in CD34+ hematopoietic progenitor cells. Specific Aims are: (1) Evaluate, optimize and assess the effects of KSHV and rKSHV.219 de novo infection of CD34+ cells in vitro. Determine if KSHV establishes a latent infection in human CD34+ hematopoietic progenitor cells and characterize the role of viral infection on suppression of hematopoiesis. (2) Determine KSHV cell tropism and tumorigenic potential by immune reconstitution of NOD/SCID mice with CD34+ HPCs infected with KSHV and rKSHV.219. Evaluate and characterize the effect of KSHV/HHV-8 infection on hematopoiesis in vivo. (3) Determine if AZT and IFNalpha induces apoptosis in primary hematopoietic cells infected with rKSHV.219. Develop the NOD/SCID-hu mouse as a pre-clinical model for therapeutic intervention to inhibit KSHV replication.

描述（由申请人提供）：体内模型对于理解病毒性疾病的致病机制以及评估对抗这些疾病的治疗方法至关重要。 KSHV 已从 AIDS 患者中发现的 B 细胞衍生淋巴瘤中分离出来，称为 PEL 和 MCD。对 KSHV 感染的初始细胞靶点以及有助于病毒在体内传播和潜伏的细胞的了解还不完全。我们推测 B 祖细胞的 KSHV 感染是肿瘤发生启动的关键过程。在 KS 患者的造血祖细胞 (CD34+) 细胞中检测到了 KSHV 基因组序列。我们已经证明 KSHV 感染人 CD34+ 细胞，并且在这些细胞体外分化产生的 CFU-GM 克隆形成集落中维持病毒基因表达。用受感染的人 CD34+ 细胞重建淋巴细胞后，在人 (CD14+) 单核细胞和 NOD/SCID-hu 嵌合小鼠的 B 淋巴细胞中检测到 KSHV 基因组 DNA 和 LANA-1 表达。尽管该模型系统不能完美代表正常人类造血和/或 KSHV 感染的所有方面，但 NOD/SCID-hu 小鼠中 KSHV 持续存在与 CD34+ 细胞感染的结合提供了一个实验系统，可解决许多问题KSHV 体内复制、潜伏期和病理生物学的研究。这种新型动物模型将用于测定 AZT 和 IFN-α 在体内抑制 KSHV 感染的功效。本提案旨在进一步完善和开发KSHV感染的NOD/SCID-hu小鼠模型，并更好地了解KSHV感染在CD34+造血祖细胞中的作用。具体目标是：（1）评估、优化和评估KSHV和rKSHV.219在体外从头感染CD34+细胞的效果。确定 KSHV 是否在人类 CD34+ 造血祖细胞中建立潜伏感染，并表征病毒感染对造血抑制的作用。 (2)通过对感染KSHV和rKSHV.219的CD34+HPC的NOD/SCID小鼠进行免疫重建来确定KSHV细胞趋向性和致瘤潜力。评估和表征 KSHV/HHV-8 感染对体内造血的影响。 (3)确定AZT和IFNα是否诱导感染rKSHV.219的原代造血细胞凋亡。开发 NOD/SCID-hu 小鼠作为抑制 KSHV 复制的治疗干预的临床前模型。