Transgenic/SCID-hu Mouse Model to Study HIV Therapeutics

高活性抗逆转录病毒疗法（HAART）并不能根除体内的HIV-1感染，因为即使经过多年的治疗，血液和淋巴组织中仍然存在携带复制能力病毒的细胞库。因此，有必要针对潜伏感染的hiv -1感染细胞开发新的治疗方法。我们建议通过开发一种新的嵌合小鼠模型来促进这些新的治疗方法的评估，该模型中填充了HIV-1感染的小鼠细胞和对HIV-1感染敏感的人类T细胞。我们开发了一种R5-tropic HIV-1JR-CSF （JR-CSF小鼠）全长原病毒克隆转基因小鼠，这种小鼠表现出血浆病毒血症，其T细胞、单核细胞和树突状细胞产生感染性HIV-1。这些小鼠的细胞通过与ROSA26小鼠杂交来标记，从而产生能够表达HIV-1JR-CSF和β -半乳糖苷酶的转基因小鼠。这些小鼠也与DO11.10 OVA (323-339)-TCR转基因小鼠杂交，允许产生含有复制能力的HIV-1的抗原特异性记忆CD4+ T细胞。我们现在证明，转移到thy/liv- SCID-hu小鼠体内的JR-CSF小鼠细胞作为hiv -1感染细胞的储存库，能够感染人类T细胞和人类胸腺植入物。只要小鼠接受HAART治疗，这种情况就不会发生，但在HAART停止后，甚至在HAART治疗1个月后，这种情况就会发生。完全清除能够产生传染性HIV-1的JR-CSF细胞将在HAART停止后防止hu-thy/liv植入物的感染。因此，我们可以通过将JR-CSF小鼠细胞转移到使用HAART的thy/liv- scid -hu小鼠体内，用候选治疗药物治疗小鼠，停止小鼠的HAART治疗，然后评估HIV-1感染在hu-thy/liv植入物中的时间发病情况，来评估针对持久性HIV-1储存库的治疗干预措施的有效性。我们将建立模型系统的有效性，并使用它来评估各种治疗方法的疗效，如环境导向毒素和HIV特异性CTL来消耗HIV-1感染细胞的群体。我们还将使用我们完善的thy/liv- SCID-hu小鼠系统，研究环境导向毒素治疗消除hiv -1感染细胞储存库的功效。

Highly active antiretroviral therapy (HAART) does not eradicate HIV-1 infection from the body because reservoirs of cells harboring replication- competent virus persist in blood and lymphoid tissue even after years of treatment. Thus, there is a need to develop new therapeutic approaches targeting the pool of latently infected HIV-1-infected cells. We propose to facilitate evaluation of these new treatments by developing a new chimeric mouse model populated with HIV-1-infectious mouse cells as well as human T cells susceptible to HIV-1 infection. We have developed mice transgenic for a full-length proviral clone of R5-tropic HIV-1JR-CSF (JR-CSF mice) that displays plasma viremia and whose T cells, monocytes and dendritic cells produce infectious HIV-1. Cells from these mice have been tagged by crossing them with ROSA26 mice to yield mice transgenic for both the expression of HIV-1JR-CSF and beta-galactosidase. These mice have also been crossed with DO11.10 OVA (323-339)-TCR transgenic mice permitting generation of antigen-- specific memory CD4+ T cells containing replication-competent HIV-1. We now demonstrate that JR-CSF mouse cells transferred into thy/liv- SCID-hu mice function as a reservoir of HIV-1-infected cells capable of infecting the human T cells and human thymic implant. This is prevented as long as the mice were treated with HAART, but occurs after HAART is stopped, even after 1 month of HAART. Complete clearance of the JR-CSF cells capable of producing infectious HIV-1 would prevent infection of the hu-thy/liv implant after HAART was stopped. Thus, we could evaluate the efficacy of therapeutic interventions to target persistent HIV-1 reservoirs by transferring JR-CSF mouse cells into thy/liv-SCID-hu mice on HAART, treating the mice with the therapeutic candidates, stopping HAART treatment of the mice, and then evaluating the temporal onset of HIV-1 infection in the hu-thy/liv implant. We will establish the validity of the model system and use it to evaluate the efficacy of various treatments such as Env-directed toxins, and HIV- specific CTL to deplete the population of HIV-1-infected cells. We will also examine the efficacy of Env-directed toxin-treatment to eliminate reservoirs of HIV-1-infected cells using our well-established thy/liv- SCID-hu mouse system.