Recombinant Protein Immunogens

双模式（也称为“初免-加强”）免疫利用了不同的反应， 由不同背景下的免疫原产生。这些包括对不同的 佐剂和载体，以及由不同抗原呈递引起的抗原特异性应答 途径。近年来，努力集中在通过使用免疫球蛋白来产生细胞介导的免疫（CMI）。 非复制型病毒载体，如MVA或复制缺陷型腺病毒，和DMA疫苗。然而，在这方面， 体液反应的重要性，包括广泛反应的中和抗体和抗体- 依赖性细胞毒性（ADCC）活性已越来越被认识到。因此有 有必要探索可能诱导CMI和体液反应的引发-加强策略。一个有吸引力 策略的例子是用可复制的病毒载体（例如，腺病毒）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 该核心的总体目标是为项目2和3的研究者提供重组亚单位 蛋白质，包括HIV-1进化枝C gp 160、SIV Gag-Pol颗粒以及重组SIVmac 239 Nef 蛋白该核心的具体目标是： (1)目的：从重组质粒中表达、鉴定和纯化HIV-1 C进化枝包膜糖蛋白gp 160， 牛痘病毒; (2)从重组牛痘病毒中表达、鉴定和纯化SIVmac 239 Gag-Pol颗粒; (3)从重组E.杆菌

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.