Induction of mucosal SIV immunity in non human primates by secreted Hsp-Gp96

通过分泌的 Hsp-Gp96 诱导非人灵长类动物粘膜 SIV 免疫

• 批准号: 7382544
• 负责人: ECKHARD R PODACK
• 金额: $ 18.6万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-15 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7382544
• 关键词: Adjuvant; Antibody Formation; Antigens; Attenuated; Binding; Biological Models; CD4 Positive T Lymphocytes; CD40 Ligand; CD8B1 gene; Cell Death; Cells; Cellular Immunity; Charge; Chimeric Proteins; Clinical Trials; Clonal Expansion; Collaborations; Complex; Cross Presentation; Cross-Priming; Cytolysis; Dendritic Cells; Dendritic cell activation; Dose; Endoplasmic Reticulum; Generations; HIV/SIV vaccine; Heat shock proteins; Human; Immune; Immune response; Immunity; Immunologist; Institution; Iron binding capacity measurement; Knowledge; Lamina Propria; Malignant neoplasm of lung; Memory; Modeling; Molecular Chaperones; Mucosal Immunity; Mus; NYVAC vaccine; Natural Killer Cells; Necrosis; Pathogenesis; Peptide Transport; Peptides; Preparation; Process; Property; Proteins; RPS19 gene; Research Ethics Committees; SIV; SIV Vaccines; Side; Signal Transduction; Simulate; Stimulus; Structure of aggregated lymphoid follicle of small intestine; T-Cell Activation; T-Lymphocyte; TLR2 gene; TLR4 gene; TNFRSF5 gene; Testing; United States Food and Drug Administration; United States National Institutes of Health; Universities; Vaccines; Viral; Viral Vaccines; Virus; Virus Diseases; base; immunogenicity; in vivo; intraepithelial; lymph nodes; lysyl-aspartyl-glutamyl-leucine; macrophage; neoplastic cell; nonhuman primate; receptor; response; success

DESCRIPTION (provided by applicant): Cellular immunity and memory is required for clearance of viruses and for protection from viral infection. Cellular immunity by CD8 CTL and NK cells is initiated through activation of the innate immune response, maturation of dendritic cells (DC) and antigen cross presentation to CD8 T cells. Anti viral vaccines stimulating cellular immunity have to imitate this process. Adjuvants for the stimulation of antibody responses are used effectively however our knowledge about adjuvants for the stimulation of CTL immunity is limited. We have demonstrated that the endoplasmic reticulum resident heat shock protein gp96, a chaperone for peptides transported to be presented by MHC-I, is a natural adjuvant for the activation of DC, NK and cognate CD8 T cells. DC and macrophages have receptors for gp96-peptide complexes and become activated upon gp96-binding. Gp96 together with its associated peptides is taken up the APC, the peptide moiety is transported to the ER and used to charge MHC-I molecules. Concomitant activation of DC results in more than million fold enhanced cross priming of cognate CD8 T cells when compared to cross priming by intact protein taken up by DC. To take advantage of this unique adjuvant effect and the ability to transport relevant peptides, we have made a secretable form of gp96, gp96-lg, by replacing the KDEL ER-retention signal with the Fc portion of lgG1. We hypothesize that cell-based gp96-lg vaccines, by prolonged in vivo secretion of gp96-lg-peptide, imitate viral replication and provide immune stimuli comparable to attenuated viruses. In model systems in mice we have shown that gp96-lg transfected, antigen expressing tumor cells secrete gp96-lg in vivo and stimulate the innate DC and NK as well as adaptive, cognate cellular CD8 CTL immune response and generate specific CD8 memory independent of CD4 help and in the absence of lymph nodes. Both systemic and strong mucosal immunity in intraepithelial, lamina propria and Peyer's patch CD8 CTL is generated by gp96-lg vaccines. Because of their unique properties we now plan to evaluate the gp96- vaccines in non-human primate models for SIV for their immunogenicity for mucosal and systemic cellular immunity (R21). In addition we will examine the protective power of SIV-gp96-vaccines against subsequent viral challenge (R33). To maximize the chances of success, the team in Miami (Podack/Pahwa) has entered into a collaboration with experts (Franchini lab) at the NIH bringing together basic immunologists with experts in human and non human primate HIV/SIV pathogenesis.

描述(申请人提供)：清除病毒和防止病毒感染需要细胞免疫和记忆力。CD8CTL和NK细胞的细胞免疫是通过激活先天免疫反应、树突状细胞(DC)的成熟和抗原向CD8T细胞的交叉递呈而启动的。刺激细胞免疫的抗病毒疫苗必须模仿这一过程。佐剂用于刺激抗体反应是有效的，但我们对佐剂用于刺激CTL免疫的了解有限。我们已经证明，内质网驻留的热休克蛋白gp96是一种天然的DC、NK和同源CD8 T细胞激活佐剂，它是MHC-I转运的多肽的伴侣。DC和巨噬细胞有gp96-肽复合体的受体，并在gp96结合后被激活。Gp96及其相关多肽被APC摄取，多肽部分被输送到内质网，用于给MHC-I分子充电。与DC摄取的完整蛋白的交叉激发相比，伴随的DC激活导致同源CD8 T细胞的交叉激发增强了数百万倍以上。为了利用这种独特的佐剂作用和运输相关多肽的能力，我们用lgG1的Fc部分取代了KDEL ER保留信号，制造了一种可分泌的gp96形式，gp96-lg。我们假设，基于细胞的gp96-lg疫苗，通过延长体内gp96-lg多肽的分泌，模拟病毒复制，并提供与减毒病毒相当的免疫刺激。在小鼠的模型系统中，我们已经证明了gp96-LG转基因的肿瘤细胞在体内分泌gp96-LG，刺激固有的DC和NK以及适应性的同源细胞CD8 CTL免疫反应，并在没有淋巴结的情况下产生不依赖于CD4帮助的特异性CD8记忆。Gp96-LG疫苗可产生上皮内、固有层和Peyer‘s斑块CD8 CTL的系统免疫和较强的粘膜免疫。由于其独特的特性，我们现在计划在非人类灵长类动物模型中评估gp96疫苗对SIV的黏膜和系统细胞免疫的免疫原性(R21)。此外，我们还将研究SIV-gp96-疫苗对后续病毒挑战的保护力(R33)。为了最大限度地增加成功的机会，迈阿密(Podack/Pahwa)的团队与NIH的专家(Franchini实验室)进行了合作，将基础免疫学家与人类和非人类灵长类艾滋病毒/SIV发病机制的专家聚集在一起。