Parsons Christopher H, Adang Laura A, Overdevest Jon, O'Connor Christine M, Taylor J Robert, Camerini David, Kedes Dean H
Myles H. Thaler Center for AIDS and Human Retrovirus Research, Department of Microbiology, University of Virginia Health Systems, Charlottesville, Virginia 22908, USA.
J Clin Invest. 2006 Jul;116(7):1963-73. doi: 10.1172/JCI27249. Epub 2006 Jun 22.
To develop an animal model of Kaposi sarcoma-associated herpesvirus (KSHV) infection uniquely suited to evaluate longitudinal patterns of viral gene expression, cell tropism, and immune responses, we injected NOD/SCID mice intravenously with purified virus and measured latent and lytic viral transcripts in distal organs over the subsequent 4 months. We observed sequential escalation of first latent and then lytic KSHV gene expression coupled with electron micrographic evidence of virion production within the murine spleen. Using novel technology that integrates flow cytometry with immunofluorescence microscopy, we found that the virus establishes infection in murine B cells, macrophages, NK cells, and, to a lesser extent, dendritic cells. To investigate the potential for human KSHV-specific immune responses within this immunocompromised host, we implanted NOD/SCID mice with functional human hematopoietic tissue grafts (NOD/SCID-hu mice) and observed that a subset of animals produced human KSHV-specific antibodies. Furthermore, treatment of these chimeric mice with ganciclovir at the time of inoculation led to prolonged but reversible suppression of KSHV DNA and RNA levels, suggesting that KSHV can establish latent infection in vivo despite ongoing suppression of lytic replication.
为了建立一种特别适合评估卡波西肉瘤相关疱疹病毒(KSHV)感染的病毒基因表达、细胞嗜性和免疫反应纵向模式的动物模型,我们通过静脉注射向NOD/SCID小鼠注入纯化病毒,并在随后的4个月内测量远端器官中的潜伏和裂解病毒转录本。我们观察到KSHV基因表达先是潜伏性的,然后是裂解性的,呈顺序性升高,同时电子显微镜证据显示在小鼠脾脏中有病毒粒子产生。使用将流式细胞术与免疫荧光显微镜相结合的新技术,我们发现该病毒在小鼠B细胞、巨噬细胞、NK细胞以及程度较轻的树突状细胞中建立感染。为了研究在这种免疫受损宿主中产生人类KSHV特异性免疫反应的可能性,我们给NOD/SCID小鼠植入功能性人类造血组织移植物(NOD/SCID-hu小鼠),并观察到一部分动物产生了人类KSHV特异性抗体。此外,在接种时用更昔洛韦治疗这些嵌合小鼠导致KSHV DNA和RNA水平受到长期但可逆的抑制,这表明尽管裂解复制持续受到抑制,KSHV仍能在体内建立潜伏感染。