Induction of Apoptosis by HIV-1 Infected Monocytic Cells

描述（由申请人提供）：在R-01授权期内，我们从CD141/低CD16/高43/HIV人巨噬细胞杂交瘤细胞系中分离出一种凋亡蛋白。凋亡蛋白对应于人类假设的蛋白FLJ21908，我们已将其重命名为SHIVA（可溶性HIV凋亡）蛋白。SHIVA可能有助于HIV-1感染患者的普遍凋亡状态，也可能在艾滋病的神经系统并发症中发挥作用，特别是HIV相关性痴呆（HAD），因为它诱导原代神经元和神经元细胞系的凋亡。SHIVA可以通过免疫组化在HAD患者的脑巨噬细胞中共定位，并且可以通过ELISA和实时PCR在HAD患者的脑脊液中检测到，但在无痴呆的HIV感染患者或正常对照中无法检测到。我们已经确定了SHIVA的候选受体（MW 100 kDa），并且还证明了SHIVA诱导的SH-SY5Y神经元细胞凋亡可以被美金刚阻滞。SHIVA在感染43Hrv细胞4周后诱导，通过激活Bad和Bax，抑制Bcl和Bcl- xl，并从线粒体释放细胞色素c，激活caspase 9和caspase 3，导致细胞凋亡。抗氧化剂和转染的Bcl-2阻断shiva诱导的细胞凋亡。根据审稿人的进一步建议，在第二次重新提交的论文中，我们将再次关注体外43HW细胞中SHIVA产生的自然史，研究参与SHIVA调节的病毒和宿主细胞因子，其次是SHIVA受体的鉴定，第三是SHIVA在HIV相关痴呆（HAD）中的作用，将SHIVA的存在与HAD和非HAD的HIV患者脑组织中的凋亡联系起来。

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.