Recombinant Protein Immunogens

双模式免疫(也称为“优质免疫”)利用了不同的反应。 由出现在不同环境中的免疫原产生。这些包括对不同的 佐剂和载体，以及不同抗原提呈引起的抗原特异性反应 小路。近年来，人们一直致力于通过使用细胞因子产生细胞免疫(CMI)。 非复制型病毒载体，如MVA或复制缺陷型腺病毒，以及DNA疫苗。然而， 体液反应的重要性，包括广泛反应的中和抗体和抗体- 依赖于细胞的细胞毒性(ADCC)活性，已得到越来越多的认识。因此，它是 有必要探索可能诱发CMI和体液反应的主要助推策略。一个有吸引力的人 策略的例证是用具有复制能力的病毒载体(例如，腺病毒)来启动表达 多个HIV抗原，然后用亚单位蛋白免疫原加强免疫，如本计划所建议的 项目。 这个核心的总体目标是为项目2和项目3的研究人员提供重组亚基 蛋白质，包括HIV-1分支C gp160、SIV Gag-Pol颗粒以及重组SIVmac239Nef 蛋白。这一核心的具体目标是： (1)重组人HIV-1囊膜糖蛋白gp160的表达、鉴定和纯化 牛痘病毒； (2)从重组痘苗病毒中表达、鉴定和纯化SIVmac239 Gag-Pol颗粒； (3)从重组大肠杆菌中表达、鉴定和纯化SIVmac239 Nef。

Bi-modal (also known as "prime-boost") immunization takes advantage of the diverse responses generated by immunogens presented in different contexts. These include innate responses to different adjuvants and vectors, as well as antigen-specific responses resulting from different antigen presentation pathways. In recent years, effort has been focused on generating cell-mediated immunity (CMI) by the use of non-replicating viral vectors, such as MVA or replication-defective adenovirus, and DMAvaccines. However, the importance of humoral responses, including broadly reactive neutralizing antibodies and antibody- dependent cellular cytotoxicity (ADCC) activities, has been increasingly recognized. Therefore, it is necessary to explore prime-boost strategies likely to induce both CMI and humoral responses. An attractive strategy is exemplified by priming with replication-competent viral vectors (e.g., adenovirus) expressing multiple HIV antigens, followed by boosting with subunit protein immunogens, as proposed in this Program Project. The overall goal of this Core is to provide investigators in Projects 2 and 3 with recombinant subunit proteins, including HIV-1 clade C gp160, SIV Gag-Pol particles, as well as recombinant SIVmac239 Nef protein. The specific aims of this Core are: (1) To express, characterize and purify clade C HIV-1 envelope glycoprotein gp160 from recombinant vaccinia viruses; (2) To express, characterize and purify SIVmac239 Gag-Pol particles from recombinant vaccinia viruses; (3) To express, characterize and purify SIVmac239 Nef from recombinant E. coli.