Engineering Primary T cells for Resistance to HIV-1

描述(由申请人提供)：这项研究的申请集中在产生抗HIV-1的初级T细胞作为潜在的HIV-1翻译免疫疗法。其具体目标包括：1)用HIV-1融合抑制物C34的膜结合形式设计原代T细胞，评估抗病毒效果和细胞免疫功能，2)开发比较和结合慢病毒骨干载体中的多靶点转基因的策略，以及3)研究由于与靶细胞上表达的融合抑制物结合而暴露的HIV表位的免疫原性。一个自我失活的慢病毒载体系统将被用来单独传递编码HIV-1进入抑制物的膜结合形式的基因，并与针对HIV-1共同受体CCR5以及HIV基因REV和TAT的小干扰RNA相结合。转导的原代T细胞将受到包括初级分离株在内的各种HIV-1病毒的挑战，并将对细胞的免疫功能进行评估。在原代T细胞中对膜结合进入抑制物的初步研究是有希望的，我们假设与针对病毒共受体和/或调节HIV基因的siRNA相结合将具有相加的抗病毒作用。这份提案描述了一项为期5年的培训计划，旨在发展儿科过敏和免疫学(A/I)的学术生涯。这位首席研究员在费城儿童医院完成了儿科住院医师培训，目前是A/I奖学金的第三个年头。这笔赠款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)和艾布拉姆森家庭癌症研究所(AFCRI)提供的各种资源。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事业发展。琼博士是AFCRI翻译研究项目的主任，领导了T细胞生物学和癌症和艾滋病毒感染的过继免疫疗法方面的开创性翻译研究。霍克西博士是宾夕法尼亚大学CFAR的主任，在艾滋病毒病毒学、发病机制和病毒进入方面拥有广泛的背景。

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.