Induction of Apoptosis by HIV-1 Infected Monocytic Cells

描述(申请人提供)：在R-01授权期内，我们从CD141/Low CD16/High 43/HIV人巨噬细胞杂交瘤细胞系中分离出一种凋亡蛋白。该凋亡蛋白对应于人类假想蛋白Flj21908，我们已将其重命名为Shiva(可溶性HIV凋亡)蛋白。Shiva可能参与了HIV-1感染患者的全身细胞凋亡状态，也可能在AIDS，特别是HIV相关性痴呆(HAD)的神经系统并发症中发挥作用，因为它诱导了原代神经元和神经细胞系的凋亡。免疫组织化学方法检测到HAD患者脑巨噬细胞中有SHIVA共定位，而HAD患者脑脊液中有SHIVA的表达，而HIV感染的非痴呆患者和正常对照组脑脊液中不存在SHIVA的表达。我们已经确定了Shiva的一个候选受体(分子量100 kDa)，并证明了在SH-SY5Y神经细胞中Shiva诱导的细胞凋亡可以被美金刚阻断。SHIVA在43Hrv细胞感染四周后被诱导，通过激活Bad和Bax，抑制Bcl和Bclxl，并从线粒体释放细胞色素c来激活caspase9和caspase3，从而引起细胞凋亡。抗氧化剂和转染的bcl2阻断了Shiva诱导的细胞凋亡。在第二次重新提交以回应评审员的进一步建议时，我们将再次关注体外43HW细胞中Shiva产生的自然历史，调查参与其调控的病毒和宿主细胞因素，第二，鉴定Shiva受体，第三，其在HIV相关性痴呆(HAD)中的作用，将Shiva的存在与有无HAD的HIV患者脑组织中的细胞凋亡联系起来。

DESCRIPTION (provided by applicant): In this R-01 granting period we have identified an apoptotic protein isolated from the CD141/low CD16/high 43/HIV human macrophage hybridoma cell line. The apoptotic protein corresponds to the human hypothetical protein FLJ21908 that we have renamed as the SHIVA (soluble HIV apoptotic) protein. SHIVA may contribute to the generalized state of apoptosis in HIV-1 infected patients and may also play a role in the neurological complications of AIDS especially HIV associated dementia (HAD) since it induces apoptosis in primary neurons and neuronal cell lines. SHIVA can be co-localized in brain macrophages of patients with HAD by immunohistochemistry and can be detected by ELISA and real time PCR in the cerebrospinal fluid of patients with HAD but not in HIV infected patients without dementia, or normal controls. We have identified a candidate receptor (MW 100 kDa) for SHIVA and have also demonstrated that SHIVA-induced apoptosis can be blocked by memantine in SH-SY5Y neuronal cells. SHIVA is induced four weeks after infection in the 43Hrv cells and causes apoptosis by activating Bad and Bax, suppressing Bcl and Bcl-xL, and releasing cytochrome c from the mitochondria that activates caspase 9 and caspase 3. Anti-oxidants and transfected Bcl-2 blocks SHIVA-induced apoptosis. In this second re-submission in response to the further recommendations of the Reviewers, we will again focus first on the natural history of SHIVA production in the 43HW cells in vitro investigating both viral and host cell factors involved in its regulation, second on the identification of a SHIVA receptor and third on its role in HIV associated dementia (HAD) correlating the presence of SHIVA with apoptosis in brain tissue of HIV patients with and without HAD.