Engineering Primary T cells for Resistance to HIV-1

描述（由申请人提供）：本研究申请的重点是产生HIV-1抗性原代T细胞，作为HIV-1的潜在翻译免疫疗法。具体目标包括：1）用膜结合形式的HIV-1融合抑制剂C34改造原代T细胞，评价抗病毒作用和细胞免疫功能，2）开发在慢病毒骨架载体中比较和联合收割机组合多靶转基因的策略，和3）研究由于与靶细胞上表达的融合抑制剂结合而暴露的HIV表位的免疫原性。自失活慢病毒载体系统将用于单独递送编码膜结合形式的HIV-1进入抑制剂的基因，以及与针对HIV-1共受体CCR 5的小干扰RNA和HIV基因Rev和达特的组合。转导的原代T细胞将用各种HIV-1病毒（包括原代分离株）进行攻击，并评价细胞的免疫功能。在原代T细胞中的初步研究对于膜结合进入抑制剂是有希望的，并且我们假设与针对病毒共受体和/或调节HIV基因的siRNA的组合将具有附加的抗病毒作用。该提案描述了一个为期5年的培训计划，用于发展儿科过敏和免疫学（A/I）的学术生涯。主要研究者已在费城儿童医院完成儿科住院医师培训，并在她的A/I奖学金的第三年。该补助金将使研究人员能够扩大她的科学技能，同时受益于宾夕法尼亚大学（UPENN）免疫学系，艾滋病研究中心（CFAR）和Abramson家庭癌症研究所（AFCRI）的各种资源。卡尔·琼博士和詹姆斯·霍克西博士将指导佩雷斯博士的科学职业发展。June博士是AFCRI转化研究项目的主任，领导了T细胞生物学和癌症和HIV感染的过继免疫疗法的开创性转化研究。Hoxie博士是宾夕法尼亚大学CFAR主任，在艾滋病毒病毒学、发病机制和病毒进入方面有着广泛的背景。

DESCRIPTION (provided by applicant): This research application focuses on the generation of HIV-1 resistant primary T cells as potential translational immunotherapies for HIV-1. The specific aims include: 1) engineering primary T cells with a membrane bound form of the HIV-1 fusion inhibitor C34, evaluation of antiviral effects and cellular immune function, 2) development of strategies to compare and combine multi-target transgenes in a lentiviral backbone vector and 3) investigation of the immunogenicity of HIV epitopes exposed as a result of binding to the fusion inhibitor expressed on the target cell. A self-inactivating lentiviral vector system will be used to deliver genes coding for membrane bound forms of an HIV-1 entry inhibitor alone, and in combination with small interfering RNAs against HIV-1 co-receptor CCR5, and HIV genes Rev and Tat. Transduced primary T cells will be challenged with a variety of HIV-1 viruses including primary isolates, and cells will be evaluated for immunologic function. Preliminary studies in primary T cells are promising for the membrane bound entry inhibitor, and we hypothesize that combinations with siRNA against viral co-receptor and/or regulatory HIV genes will have an additive anti-viral effect. This proposal describes a 5-year training program for the development of an academic career in Pediatric Allergy and Immunology (A/I). The principal investigator has completed Pediatric residency training at the Children's Hospital of Philadelphia, and is in her third year of A/I Fellowship. This grant will allow the investigator to broaden her scientific skills while benefiting from the diverse resources available at the University of Pennsylvania (UPENN) within the Department of Immunology, the Center for AIDS Research (CFAR), and the Abramson Family Cancer Research Institute (AFCRI). Drs. Carl June and James Hoxie will mentor Dr. Perez's scientific career development. Dr. June is the Director of the Translational Research Program of the AFCRI, and has led pioneering translational research in T cell biology and adoptive immunotherapy of cancer and HIV infection. Dr. Hoxie, Director of the UPENN CFAR, has an extensive background in HIV virology, pathogenesis and viral entry.