AIDS-Restrictive Innate Immune Mechanisms

描述(申请人提供)：HIV-1疾病是一个严重的国际卫生问题，也是撒哈拉以南非洲的主要死亡原因。治疗性和预防性治疗在可获得性和有效性方面是有限的，因此非常需要对控制这种病毒的替代方法进行关键评估。该项目的总体目标是阐明撒哈拉以南非洲对艾滋病毒-1疾病自然产生的保护性免疫的机制。其具体目的是(1)确定NK细胞在减少母婴传播和减缓成人HIV-1疾病进展为艾滋病中的作用；(2)确定人类白细胞抗原-Bw4 80！通过媒介预防艾滋病毒传播和疾病进展。在第一个目标中，我们将进行一项横断面研究，收集艾滋病毒传播者和非传播者母亲的外周血样，通过产生抗病毒可溶性因子(如干扰素-γ、MIP-1a&(3和RANTES)以及细胞杀伤试验)来评估他们对艾滋病毒感染的自身和儿童来源的T细胞的NK细胞反应。我们还将对病毒控制者和非病毒控制者的HIV-1感染者的血液样本进行类似的分析。为了达到第二个目的，我们将产生表达NFAT-LacZ和NK细胞受体KIR3DS1的报告细胞。我们将通过与HIV-1感染和未感染的靶细胞株表达假定的配体-B57的体外共培养试验来评估KIR3DS1对HLA-B的识别。为了进一步探索KIR3DS1识别的分子机制，将采用人类白细胞抗原突变或HIV多肽筛选。这项研究计划非常适合候选人威廉·卡尔的职业目标，他将成为一名国际卫生研究的独立研究员，专门研究先天性病毒免疫。拟议的职业发展计划将包括在美国由Marcus Altfeld博士和Bruce Walker博士担任导师，在南非由Hoosen Coovardia博士和Salim Abdool Karim博士担任导师的严密指导的基础科学研究，南非德班多丽丝·杜克研究中心和美国马萨诸塞州总医院合作伙伴艾滋病研究中心的最先进的研究设施，以及国际研究中的伦理教学课程。这一计划非常适合卡尔博士在南非建立长期的科学合作关系，并确立自己作为国际卫生研究的独立研究员的地位

DESCRIPTION (provided by applicant): HIV-1 disease is a serious international health problem and a major cause of death in sub-Saharan Africa. Therapeutic and preventative treatments are limited in availability and effectiveness, thus critical evaluations of alternative approaches to control this virus are greatly needed. The overall goal of this project is to elucidate the mechanisms underlying naturally occurring protective immunity to HIV-1 disease in sub- Saharan Africa. The specific aims are (1) to determine the role of NK-cells in decreasing mother to child HIV- 1 transmission and in slowing HIV-1-disease progression to AIDS in adults and (2) to determine the molecular mechanisms of HLA-Bw4 80! mediated protection from HIV transmission and disease progression. In the first aim we will conduct a cross-sectional study by collecting peripheral blood samples from mothers who are HIV-transmitters versus non-transmitters and assessing their NK-cell responses to HIV-infected self and child-derived T cells by the production of anti-viral soluble factors (e.g., interferon-y, MIP-1a & (3, and RANTES) as well as by cell-killing assays. We will also conduct a similar analysis with blood samples from HIV-1-infected adults that are virus controllers compared to virus non-controllers. To address the second aim we will generate reporter cells expressing NFAT-LacZ and the NK cell receptor, KIR3DS1. We will assess HLA-B recognition by KIR3DS1 by in vitro co-culture assays with HIV-1 infected and uninfected target cell lines expressing the putative ligand, HLA-B57. To probe molecular mechanisms of recognition by KIR3DS1 further, HLA mutagenesis or HIV peptide screening will be employed. The research plan is ideal for the career objective of the candidate, William Carr, to become an independent investigator in international health research, specializing in innate viral immunity. The proposed career development plan will consist of closely mentored basic-science research with Drs. Marcus Altfeld and Bruce Walker as mentors in the U.S. and Drs. Hoosen Coovardia and Salim Abdool Karim as a mentors in South Africa, state-of-the-art research facilities at the Doris Duke Research Center in Durban, South Africa, and at the Partners AIDS Research Center at Massachusetts General Hospital in the U.S., and a didactic curriculum on ethics in international research. This plan is well suited for Dr. Carr to establish long-term scientific collaborations in South Africa and to establish himself as an independent investigator in international health research