Modeling KSHV Infection and Pathogenesis in SCID Mice

SCID 小鼠 KSHV 感染和发病机制建模

• 批准号: 7140175
• 负责人: GEROLD FEUER
• 金额: $ 15.96万
• 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/7140175
• 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; 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）：体内模型对于了解病毒疾病中的致病机制以及评估治疗方法对抗这些疾病至关重要。 KSHV已从B细胞衍生的淋巴瘤中分离出来，在艾滋病患者（称为PEL和MCD）中。对KSHV感染的初始细胞靶标和细胞的初始细胞靶标有不完全理解，这些细胞在体内有助于病毒传播和潜伏期。我们推测，祖细胞B细胞的KSHV感染是造成肿瘤发生的关键过程。在KS患者的造血祖细胞（CD34+）细胞中已经检测到KSHV基因组序列。我们已经证明了KSHV感染了人CD34+细胞，并且病毒基因表达保持在CFU-GM克隆菌落中，这些菌落从这些细胞的分化中得出。在与感染人类CD34+细胞的淋巴细胞重建后，在人（CD14+）单核细胞和BON/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。尽管该模型系统并不能完全代表正常的人类造血和/或KSHV感染的所有方面，但在NOD/SCID-HU小鼠中的KSHV持久性与CD34+细胞的感染的结合提供了一个实验系统，可以解决KSHV复制，延迟，延迟和培养基在VIVO中的问题。这种新型动物模型将用于测定AZT和IFN-α在抑制体内KSHV感染中的功效。该建议旨在进一步完善和开发KSHV感染的点头/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。具体目的是：（1）在体外评估，优化和评估CD34+细胞从头感染KSHV和RKSHV的影响。确定KSHV是否在人CD34+造血祖细胞中建立潜在感染，并表征病毒感染在抑制造血的作用。 （2）通过用KSHV和RKSHV.219感染的CD34+ HPC对NOD/SCID小鼠的免疫重建来确定KSHV细胞的近端和肿瘤潜能。评估和表征KSHV/HHV-8感染对体内造血的影响。 （3）确定AZT和IFNALPHA是否诱导了原发性造血细胞的凋亡，感染了RKSHV.219。开发点头/scid-hu小鼠作为治疗干预措施的临床前模型，以抑制KSHV复制。