Neisserial Porins and Antigen Presenting Cells

奈瑟氏球菌孔蛋白和抗原呈递细胞

• 批准号: 8707307
• 负责人: LEE Mark WETZLER
• 金额: $ 56.44万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707307
• 关键词: Adaptor Signaling Protein; Adjuvant; Adjuvanticity; Antibodies; Antibody Affinity; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Breeding; CD4 Positive T Lymphocytes; CD8B1 gene; Cells; Cellular Immunity; Cytotoxic T-Lymphocytes; DNA; Data; Dendritic Cells; Immune; Immune response; Immunobiology; Immunoglobulin G; Immunologic Adjuvants; Infection; Laboratories; Licensing; Ligands; Listeria; Listeria monocytogenes; Listeriosis; MHC Class II Genes; Measures; Mediating; Microbiology; Modeling; Mus; Nature; Ovalbumin; Pattern recognition receptor; PorB porin; Property; Proteins; Role; Saponin; Saponins; Signal Transduction; T cell response; T-Lymphocyte; TLR1 gene; TLR4 gene; TLR6 gene; Testing; Vaccine Adjuvant; Vaccines; Work; aluminum sulfate; cell motility; immune activation; lymph nodes; macrophage; medical schools; monophosphoryl lipid A; pathogen; porin; prevent; public health relevance; receptor; recombinase; research study; response; vaccine candidate

DESCRIPTION (provided by applicant): Our laboratory has been investigating the immunobiology of the Neisserial porin PorB for the last two decades. We first began by investigating the use of this protein as a potential anti-Neisserial candidate, but through this work we found that this protein had unique immune stimulatory abilities above and beyond its own potential use as a vaccine. We found that PorB can be used as an immune adjuvant for vaccines and that this ability was directly related to its ability to stimulate antigen presenting cells (APC) and increase expression of the costimulatory molecule CD86 and class II MHC. Moreover, we found that the ability of PorB to stimulate and activate antigen presenting cells was directly due to an interaction with the pattern recognition receptor, TOLL-like Receptor 2 (TLR2) and requires the TLR adaptor protein MyD88. We have further shown, as described in the preliminary data section and in that TLR2 needs to heterodimerize with TLR1 (and not TLR6) in order for PorB to stimulate these APCs. Utilizing this body of information, the next logical step for studying the immune stimulating adjuvant activity of PorB is to compare this activity to other known vaccine adjuvants. This will allow for the better use of PorB as adjuvant, by understanding what type of immune response it induces as compared to these other adjuvants. Moreover, we believe it is essential to continue to investigate the adjuvant potential of PorB, as it is currently the only TLR2 ligand being investigated as a vaccine adjuvant. We shall compare the immune stimulation by PorB to panel of other adjuvants, including the only licensed adjuvant, Alum (inflammasomes mediated immune activation, other TLR ligands, including CpG DNA (TLR9), and monophosphoryl lipid A (MPL)(TLR4), and QS-21. We shall examine the ability of these adjuvants to induce immune responses to a test antigen, ovalbumin, which we have used previously as described in the Preliminary Data section. We shall measure antibody levels induced, isotypes and IgG subtypes induced, antibody affinity, CD4 T cell response and ability to induce APC migration and lymph node specific T cell stimulation. Moreover, we shall compare the ability of these different adjuvants to protect mice from infection with Listeria which secretes OVA. This is a well characterized model and has been used previously to demonstrate that the presence of OVA specific CD4 and CD8 T cells are necessary for protection of mice from infection with this pathogen when it is constructed to secrete OVA. We shall obtain this strain from Dr. Darren Higgins in the Dept. of Microbiology at the Harvard Medical School, who has significant expertise in this model and will act as a consultant on this proposal. Even though the protection induced by most licensed vaccines are due to the induction of neutralizing or functional antibodies, we feel it would extremely important to begin to definitively compare the ability of these adjuvants, including PorB, to not only induce functional antibodies, but to potentially induce a protective CD8 CTL response.

描述（由申请人提供）：在过去的二十年中，我们的实验室一直在调查奈瑟氏孔孔蛋白Porb的免疫生物学。我们首先研究了该蛋白作为潜在的抗日日候选者的使用，但是通过这项工作，我们发现该蛋白质具有独特的免疫刺激能力，超出了其自身作为疫苗的潜在用途。我们发现，PORB可以用作疫苗的免疫佐剂，并且该能力与其刺激抗原呈递细胞（APC）的能力直接相关，并增加了共刺激分子CD86和II类MHC的表达。此外，我们发现，PORB刺激和激活抗原呈递细胞的能力直接是由于与模式识别受体Toll样受体2（TLR2）的相互作用，并且需要TLR适配器蛋白MYD88。如初步数据部分所述，我们进一步显示了TLR2需要与TLR1（而不是TLR6）进行异二聚体，以使PORB刺激这些APC。 利用这一信息，研究PORB的免疫刺激辅助活性的下一个逻辑步骤是将这种活性与其他已知的疫苗佐剂进行比较。通过了解与其他辅助物相比，它可以更好地利用PORB作为辅助物。此外，我们认为必须继续研究PORB的辅助潜力，因为它目前是唯一被研究为疫苗辅助的TLR2配体。我们将PORB与其他佐剂组的免疫刺激进行比较，包括唯一的许可辅助，校友（炎性症状体介导的免疫激活，其他TLR配体，包括CpG DNA（TLR9）（TLR9）和单磷酸化脂质脂质A（MPL）（MPL4）（MPL4）（TLR4）和QS-21的能力。抗原，我们先前在“初步数据”部分中所述使用的抗原蛋白，我们应诱导的抗体水平，同类和IgG亚型，抗体亲和力，CD4 T细胞响应和诱导APC的迁移和淋巴结刺激的能力是一个表征良好的模型，以前已被用来证明OVA特异性CD4和CD8 T细胞的存在对于保护小鼠在构造以分泌OVA时免受感染的感染是必要的。我们将从哈佛医学院的微生物学系的达伦·希金斯（Darren Higgins）博士那里获得这种压力，他在该模型中具有重要的专业知识，并将担任该提案的顾问。即使大多数许可疫苗引起的保护是由于诱导中和或功能性抗体的诱导，我们认为开始确定地比较这些辅助药物（包括PORB）的能力将不仅诱导功能性抗体，而且可以潜在地诱导保护性CD8 CTL响应。