In situ gene-modified DCs for an HIV-1 T cell vaccine

用于 HIV-1 T 细胞疫苗的原位基因修饰 DC

• 批准号: 6719074
• 负责人: Jerry L Blackwell
• 金额: $ 6.11万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-15 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6719074
• 关键词: AIDS vaccines; Adenoviridae; CD40 molecule; T lymphocyte; biotechnology; cell migration; cellular immunity; dendritic cells; female; genetic transduction; human immunodeficiency virus 1; laboratory mouse; leukocyte activation /transformation; vector vaccine; AIDS vaccines; Adenoviridae; CD40 molecule; T lymphocyte; biotechnology; cell migration; cellular immunity; dendritic cells; female; genetic transduction; human immunodeficiency virus 1; laboratory mouse; leukocyte activation /transformation; vector vaccine

DESCRIPTION (provided by applicant): Although protective vaccination strategies have been developed against many infectious diseases of viral origin, a vaccine with broad activity against HIV-1 has remained elusive for two decades. Cell-mediated immunity has been implicated in (i) modulation of HIV-1 replication, (ii) protraction of disease progression, and (iii) protection against infection, arguing their importance as components of a successful immunization strategy. Dendritic cells (DC) can present antigens via MHC class I and II pathways and are central to the development of cell-mediated immune responses. Thus, DC-based vaccines are currently being developed as agents to elicit strong, antigen-specific T cell responses, but these protocols usually require ex vivo genetic modification of DCs, followed by re-infusion back into an autologous or MHC-matched host where they mature, migrate to lymph nodes, and present the candidate antigen to CD4 and/or CD8 T cells to elicit antigen-specific responses. Although the ex vivo genetic manipulation approach has been useful in establishing proof-of-concept in an experimental setting, it is not practical for the development of prophylactic vaccines to be used in the field. If transduction of the DCs could be accomplished in situ, the broad use of DC-targeted vaccines as prophylactic agents would be more feasible. However, a critical component of in situ transduction is efficient targeting of the vector to DCs without perturbation of DC function. To this end, we have generated an adenovirus (Ad) vector system that specifically targets DCs via the CD40 receptor and have shown that the CD40-targeted Ad efficiently transduces DCs in vitro without interfering with DC function. A potential limitation of in situ transduction is that sufficient numbers of DC cells must be at the vaccination site to achieve effective immunization. Therefore, we propose to recruit DCs into subcutaneous locales prior to CD40-targeted vaccination by using a novel technology, chemoattractant-releasing polymer rods. In this proposal, we will evaluate the potency of CD40-targeted Ad vectors that express HIV-1 antigens in murine in vitro and in situ vaccination models that incorporate chemoattractant-releasing polymers. Initially, we will optimize the parameters of in situ DC recruitment and determine the degree to which this contributes to the efficiency of in situ CD40-targeted transduction. Next, antigen-specific CD4 and CD8 T cell responses will be compared following immunization with untargeted and CD40-targeted Ad vectors in the presence or absence of DC recruitment. Our hypothesis is that the CD40-targeted Ad will optimally transduce DCs following subcutaneous recruitment to the immunization locale, resulting in the induction of robust T cell immunity. The ability to achieve targeted in situ immunization of DCs followed by the induction of robust antigen-specific T cell immunity will be a milestone in HIV-1 vaccine development. The success of this study will be the impetus for future in vivo studies in non-human primate models to show protective immunity and human clinical trials for populations at high risk of HIV-1 infection.

描述（由申请人提供）：尽管已经针对许多病毒起源的传染病制定了保护性疫苗接种策略，但针对HIV-1的广泛活性的疫苗持续了二十年。 细胞介导的免疫与（i）调节HIV-1复制，（ii）疾病进展的突出和（iii）免受感染的保护，认为它们是成功免疫策略的组成部分的重要性。 树突状细胞（DC）可以通过MHC I类和II途径呈现抗原，对于细胞介导的免疫反应的发展至关重要。 Thus, DC-based vaccines are currently being developed as agents to elicit strong, antigen-specific T cell responses, but these protocols usually require ex vivo genetic modification of DCs, followed by re-infusion back into an autologous or MHC-matched host where they mature, migrate to lymph nodes, and present the candidate antigen to CD4 and/or CD8 T cells to elicit antigen-specific responses. 尽管离体遗传操纵方法对于在实验环境中建立概念概念很有用，但在该领域使用预防性疫苗的开发是不切实际的。 如果可以原位完成DC的转导，则广泛使用DC靶向疫苗作为预防剂将更可行。 但是，原位转导的关键组成部分是将向量靶向DC，而无需扰动直流功能。 为此，我们生成了一个腺病毒（AD）矢量系统，该系统通过CD40受体专门针对DC，并表明CD40靶向的AD在体外有效地传递了DC，而不会干扰DC功能。 原位转导的潜在局限性是，足够数量的DC细胞必须在疫苗接种部位才能实现有效的免疫。 因此，我们建议通过使用一种新技术，趋化剂释放聚合物杆在CD40靶向疫苗之前将DC招募到皮下部分地区。 在此提案中，我们将评估CD40靶向的AD载体的效力，这些AD载体在体外和原位疫苗接种模型中表达HIV-1抗原，这些模型融合了介质吸收剂释放聚合物。 最初，我们将优化原位DC募集的参数，并确定这有助于原位CD40靶向转导的效率的程度。 接下来，在存在或不存在DC募集的情况下，将比较抗原特异性的CD4和CD8 T细胞反应与未靶向和CD40靶向的AD载体后，将比较未靶向和CD40的AD载体。 我们的假设是，靶向CD40的AD将在皮下募集后将DC最佳地转移到免疫部位，从而导致稳健的T细胞免疫诱导。 达到DCS的原位免疫接种的能力，然后诱导健壮的抗原特异性T细胞免疫，将是HIV-1疫苗发育的一个里程碑。 这项研究的成功将是非人类灵长类动物模型中未来体内研究的动力，以显示针对HIV-1感染高风险的人群的保护性免疫和人类临床试验。