Attenuated RhCMV Delta 10 SIV Oral Vaccine Vectors Encoding TLR5 Ligand Sequences

编码 TLR5 配体序列的减毒 RhCMV Delta 10 SIV 口服疫苗载体

• 批准号: 8383058
• 负责人: Peter A Barry
• 金额: $ 61.33万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2015-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8383058
• 关键词: Adjuvant; Agonist; Antibody Formation; Antigen Presentation; Antigens; Attenuated; Capsid; Capsid Proteins; Cell Maturation; Cells; Chimeric Proteins; Code; Cytomegalovirus; Cytomegalovirus Vaccines; Data; Dendritic Cells; Dendritic cell activation; Engineering; Epithelium; Female; Flagellin; Future; Generations; Genes; HIV; HIV vaccine; HIV-1; Heterophile Antigens; Human; IL10 gene; Immune; Immune response; Immune system; Immunity; Immunization; In Vitro; Infection; Inflammation; Interleukin-10; Investigation; Laboratories; Length; Ligands; Lymphoid Tissue; M cell; Macaca mulatta; Modeling; Mucosal Immune Responses; Mucous Membrane; Nature; Oral; Oral cavity; Oral mucous membrane structure; Pathogenicity; Recombinants; Route; SIV; Safety; Salmonella enterica; Serum; Site; System; TLR2 gene; TLR5 gene; Target Populations; Testing; Tissues; Toll-Like Receptor 5; Toll-like receptors; Vaccine Antigen; Vaccine Design; Vaccines; Vagina; Viral; Viral Vaccines; Viral Vector; Work; base; immunogenicity; improved; in vivo; mucosal site; mucosal vaccine; mutant; nonhuman primate; novel; novel vaccines; oral vaccine; peripheral blood; public health relevance; receptor; response; transmission process; vaccine development; vaccine efficacy; vector; vector vaccine

DESCRIPTION (provided by applicant): To explore a unique approach for generating robust long term immune responses at both mucosal and systemic lymphoid tissues in the nonhuman primate model for human immunodeficiency virus (HIV), we propose to utilize an attenuated rhesus cytomegalovirus (RhCMV) vaccine vector developed in our laboratories for oral delivery of a simian immunodeficiency virus (SIV) immunogen engineered as a fusion protein to include either full length coding sequence or specific domains of flagellin, the agonist for toll-like receptor (TLR)5. TLR5 is a potent activator of mucosal immune responses through activation of dendritic cells (DC), follicle-associated epithelium (FAE) and M cells included within gut and oral cavity mucosal- associated lymphoid tissue (MALT). As such, flagellin has proven to be a potent adjuvant for i) induction of serum and/or secretory antibody responses and for ii) induction of Th1 and Th2 responses in both systemic and mucosal tissues. This oral vaccine approach combines the advantages of a potent TLR adjuvant with a viral vector (CMV) shown to disseminate throughout the host after oral delivery/transmission and to establish a lifelong persistence in the presence of immune responses that limit viral pathogenicity at multiple sites within the host. Furthermore, our attenuated RhCMV vector has been modified by deletion of the RhCMV-encoded viral interleukin (IL)-10 gene (RhCMV IL10) that results in increased inflammation at the site of primary infection in vivo, an abrogation of the inhibition of DC maturation associated with wild type CMV infection in vitro, and attenuated viral replication for safety. Lastly, data has shown that prior immunity to human CMV (HCMV) does not preclude re-infection, implying that a CMV-based vector system may be broadly applied to a vaccine target population irrespective of pre-vaccine immunity to HCMV. We hypothesize that RhCMV IL10 vectors expressing SIV vaccine immunogens delivered orally and systemically will induce superior vaccine antigen-specific cellular and humoral immune responses in multiple mucosal sites. Also, incorporation of TLR5 agonist adjuvant activity by fusion of flagellin sequences with vaccine antigen will further enhance both innate and adaptive mucosal immune responses and provide a novel oral mucosal vaccine approach for protection against HIV. Accordingly, immunogens (SIVmac239 Capsid) fused with full length and/or specific domains of flagellin will be constructed to generate a vaccine antigen with optimal immunogenicity for mucosal tissues and will be used for generation of candidate RhCMV IL10Capsid/flagellin vaccine vectors (Aim1). Candidate RhCMV IL10Capid/flagellin vaccines will next be tested for immunogenicity (Aim 2) in multiple mucosal and systemic tissues and for efficacy (Aim 3) in RhCMV-seronegative female rhesus macaques by vaginal SIVmac251 challenge. We propose that these novel vectors carry the potential to significantly impact HIV vaccine design.

描述（由申请人提供）：为了探索一种在人类免疫缺陷病毒（HIV）的非人灵长类动物模型中在粘膜和全身淋巴组织产生强大的长期免疫反应的独特方法，我们建议利用我们实验室开发的减毒恒河猴巨细胞病毒（RhCMV）疫苗载体来口服递送猿类免疫缺陷病毒 (SIV) 免疫原被设计为融合蛋白，包括全长编码序列或鞭毛蛋白的特定结构域，鞭毛蛋白是 Toll 样受体 (TLR)5 的激动剂。 TLR5 通过激活肠道和口腔粘膜相关淋巴组织 (MALT) 内的树突状细胞 (DC)、滤泡相关上皮 (FAE) 和 M 细胞，是粘膜免疫反应的有效激活剂。因此，鞭毛蛋白已被证明是一种有效的佐剂，用于 i) 诱导血清和/或分泌性抗体反应，以及 ii) 诱导全身和粘膜组织中的 Th1 和 Th2 反应。这种口服疫苗方法结合了有效的 TLR 佐剂和病毒载体 (CMV) 的优点，经证明在口服递送/传播后可在整个宿主中传播，并在存在免疫反应的情况下建立终生持久性，从而限制宿主体内多个位点的病毒致病性。此外，我们的减毒RhCMV载体通过删除RhCMV编码的病毒白细胞介素（IL）-10基因（RhCMV IL10）进行了修饰，导致体内原发感染部位炎症增加，消除了与体外野生型CMV感染相关的DC成熟抑制，并减弱了病毒复制以确保安全。最后，数据显示，先前对人 CMV (HCMV) 的免疫力并不能排除再次感染，这意味着基于 CMV 的载体系统可以广泛应用于疫苗目标人群，而与疫苗前对 HCMV 的免疫力无关。我们假设口服和全身递送的表达SIV疫苗免疫原的RhCMV IL10载体将在多个粘膜位点诱导优异的疫苗抗原特异性细胞和体液免疫反应。此外，通过鞭毛蛋白序列与疫苗抗原融合来掺入TLR5激动剂佐剂活性将进一步增强先天性和适应性粘膜免疫反应，并提供一种新的口腔粘膜疫苗方法来预防HIV。因此，将构建与鞭毛蛋白全长和/或特定结构域融合的免疫原（SIVmac239衣壳），以生成对粘膜组织具有最佳免疫原性的疫苗抗原，并将用于生成候选RhCMV IL10衣壳/鞭毛蛋白疫苗载体（Aim1）。接下来将测试候选 RhCMV IL10Capid/鞭毛蛋白疫苗在多个粘膜和全身组织中的免疫原性（目标 2），以及通过阴道 SIVmac251 攻击在 RhCMV 血清阴性雌性恒河猴中测试功效（目标 3）。我们认为这些新型载体有可能显着影响艾滋病毒疫苗的设计。