Induction of Apoptosis by HIV-1 Infected Monocytic Cells

描述（由申请方提供）：在本R-01授予期内，我们鉴定了从CD 141/低CD 16/高43/HIV人巨噬细胞杂交瘤细胞系中分离的凋亡蛋白。凋亡蛋白对应于人类假设蛋白FLJ 21908，我们将其重命名为SHIVA（可溶性HIV凋亡）蛋白。SHIVA可能导致HIV-1感染患者的细胞凋亡的普遍状态，并且也可能在AIDS的神经系统并发症，特别是HIV相关痴呆（HAD）中起作用，因为它诱导原代神经元和神经元细胞系的细胞凋亡。免疫组化结果显示SHIVA共定位于HAD患者脑巨噬细胞中，ELISA和真实的时间PCR检测HAD患者脑脊液中SHIVA的表达，但在无痴呆的HIV感染患者和正常对照中均未检测到SHIVA。我们已经确定了一个候选受体（分子量100 kDa）SHIVA，也证明了SHIVA诱导的细胞凋亡可以阻断SH-SY 5 Y神经元细胞中的美金刚。SHIVA在感染43 Hrv细胞四周后被诱导，并通过激活Bad和Bax、抑制Bcl和Bcl-xL以及从线粒体释放细胞色素c（激活caspase 9和caspase 3）来引起细胞凋亡。抗氧化剂和转染的Bcl-2阻断SHIVA诱导的细胞凋亡。在第二次重新提交以响应审评员的进一步建议中，我们将再次首先关注体外43 HW细胞中SHIVA产生的自然历史，研究参与其调节的病毒和宿主细胞因素，第二个是关于SHIVA受体的鉴定，第三个是关于它在HIV相关痴呆（HAD）中的作用。将SHIVA的存在与有和无HAD的HIV患者脑组织中的细胞凋亡相关联。

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.