Genetically Engineered Oral Vaccines and Caries Immunity

描述（由申请人提供）：正在研究重组无毒沙门氏菌疫苗表达异源病原体毒力抗原克隆基因的能力，以及其在开发保护性对抗各种感染性疾病（包括龋齿）的粘膜疫苗中的潜在用途。然而，这些安全的活载体疫苗介导宿主应答的机制仍然知之甚少。模式识别受体（PRR），包括Toll样受体（TLR），在宿主细胞上/中表达，并识别与微生物组分相关的不同分子模式。树突状细胞上TLR对微生物组分的识别导致先天免疫应答的诱导和适应性免疫的激活。我们从表达编码变形链球菌粘附AgI/II的唾液结合区（SBR）的基因的沙门氏菌载体疫苗的免疫研究中的发现表明，共刺激分子CD 80和CD 86在介导粘膜和全身抗体应答中发挥不同的以及冗余的作用，从而提供了载体和SBR可能以不同的方式向先天免疫系统发出信号的证据，可能是通过TLR。其他证据表明，不止一种TLR可能参与介导和增强对沙门氏菌载体产生的SBR的免疫应答。我们提出的研究的基本假设是，暴露于沙门氏菌载体疫苗后参与的各种宿主细胞上的TLR之间的相互作用将影响对沙门氏菌表达的克隆抗原的适应性免疫应答的结果。我们的研究将集中在确定TLR的调节作用，介导的适应性免疫反应后，粘膜免疫与复杂的沙门氏菌载体疫苗表达克隆的沙门氏菌毒力抗原。变异人具体目标是：1）确定TLR和TLR之间的相互作用，这些TLR参与选择性增强对S克隆抗原的粘膜和全身应答。由沙门氏菌载体疫苗表达的变形杆菌，和2）确定TLR在T细胞和B细胞上对于由沙门氏菌载体疫苗表达的克隆抗原的适应性免疫应答的作用。体内免疫应答和过继转移模型， 使用TLR敲除小鼠，以及体外测定系统将用于解决的目标。了解每种TLR和参与介导对疫苗候选物的先天性和适应性宿主应答的TLR之间的相互作用，以及树突状细胞、T细胞和B细胞中TLR信号传导对产生的对特定微生物抗原的适应性应答的贡献，将有助于设计改进的沙门氏菌载体疫苗和免疫方案，用于诱导针对感染性疾病的保护性免疫应答，包括龋齿。 公共卫生相关性：重组减毒沙门氏菌疫苗正在开发和测试其在诱导对各种感染因子（包括龋齿和牙周病的病原体）的保护性粘膜和全身免疫应答中的有效性。活载体疫苗在诱导针对微生物病原体的粘膜免疫应答方面更有效，并且比由纯化的微生物组分组成的疫苗生产更经济。因此，确定宿主识别沙门氏菌载体疫苗的微生物组分的机制是至关重要的，所述沙门氏菌载体疫苗的微生物组分调节对异源病原体（例如变异链球菌）的表达克隆毒力抗原诱导的免疫应答的性质，以开发高度有效地增强针对粘膜病原体的保护性免疫应答的重组无毒沙门氏菌疫苗，包括与口腔相关的那些。

DESCRIPTION (provided by applicant): Recombinant avirulent Salmonella vaccines are being studied for their ability to express cloned genes of virulence antigens of heterologous pathogens, and for their potential use in the development of mucosal vaccines protective against various infectious diseases including dental caries. However, the mechanism(s) by which these safe, live vector vaccines mediate host responses are still poorly understood. Pattern-recognition receptors (PRRs), including the Toll-like receptors (TLRs), are expressed on/in host cells and recognize distinct molecular patterns associated with microbial components. The recognition of microbial components by TLRs on dendritic cells leads to the induction of an innate immune response and the activation of adaptive immunity. Our findings from immunization studies with a Salmonella vector vaccine expressing the gene encoding the saliva-binding region (SBR) of the adhesion AgI/II of Streptococcus mutans showed that the costimulatory molecules CD80 and CD86 play distinct, as well as redundant roles in mediating mucosal and systemic antibody responses, thus providing evidence that the vector and SBR may be signaling the innate immune system differently, presumably via TLRs. Other evidence suggests that more than one TLR may be involved in mediating and potentiating the immune response to SBR produced by the Salmonella vector. The fundamental hypothesis of our proposed studies is that the interplay between TLRs on various host cells that are engaged following exposure to the Salmonella vector vaccine will influence the outcome of the adaptive immune response to the cloned antigen expressed by the Salmonella. Our studies will focus on defining the regulatory roles of TLRs that mediate the adaptive immune response following mucosal immunization with a complex Salmonella vector vaccine expressing a cloned virulence antigen of S. mutans. The specific aims are; 1) to define the TLRs and the interplay between the TLRs involved in a selective augmentation of mucosal and systemic responses to the cloned antigen of S. mutans expressed by a Salmonella vector vaccine and 2) to determine the role of TLRs on T cells and B cells for an adaptive immune response to a cloned antigen expressed by a Salmonella vector vaccine. In vivo immune response and adoptive transfer models that use TLR knockout mice, as well as in vitro assay systems will be used to address the aims. An understanding of each TLR and the interplay between TLRs involved in mediating the innate and adaptive host responses to a vaccine candidate, and the contribution of TLR signaling in dendritic, T and B cells for the resulting adaptive response to specific microbial antigens will help in the design of improved Salmonella vector vaccines and immunization regimens for the induction of protective immune responses against infectious diseases, including dental caries. PUBLIC HEALTH RELEVANCE: Recombinant attenuated Salmonella vaccines are being developed and tested for their effectiveness in inducing protective mucosal and systemic immune responses to a variety of infectious agents, including the etiologic agents of dental caries and periodontal disease. Live vector vaccines are more effective in inducing mucosal immune responses against microbial pathogens and more economical to produce than vaccines consisting of purified microbial components. Therefore, it is critical to determine the mechanisms involved in host recognition of microbial components of a Salmonella vector vaccine that regulate the nature of the immune response induced to the expressing cloned virulence antigen of a heterologous pathogen, such as mutans streptococci, in order to develop recombinant avirulent Salmonella vaccines that are highly effective in potentiating protective immune responses against mucosal pathogens, including those associated with the oral cavity.