Neisserial Porins and Antigen Presenting Cells

奈瑟氏球菌孔蛋白和抗原呈递细胞

• 批准号: 7886100
• 负责人: LEE Mark WETZLER
• 金额: $ 56.99万
• 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/7886100
• 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; Cytotoxic T-Lymphocytes; DNA; Data; Dendritic Cells; Immune; Immune response; Immunity; 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; Toll-Like Receptor 2; Vaccine Adjuvant; Vaccines; Work; aluminum sulfate; cell motility; immune activation; lymph nodes; macrophage; medical schools; monophosphoryl lipid A; pathogen; porin; prevent; public health relevance; 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. PUBLIC HEALTH RELEVANCE: We have shown that the Neisserial porin, PorB, is a potent immune adjuvant and works via recognition by TLR2 and induction of antigen presenting cell activation. This proposal will examine the adjuvant activity of PorB and compare this activity to other currently investigated adjuvants (including Alum, CpG DNA, MPL and QS-21). We shall examine induction of both humoral responses and CD8 T cell responses and the ability of the vaccines to prevent Listeria infection in mice. Moreover, we shall further examine the mechanism of PorB adjuvant activity via the use of MyD88 floxed mice bred with mice expressing cell specific cre recombinase.

描述（由申请人提供）：过去二十年来，我们的实验室一直在研究奈瑟菌孔蛋白 PorB 的免疫生物学。我们首先研究了这种蛋白质作为潜在的抗奈瑟氏菌候选物的用途，但通过这项工作，我们发现这种蛋白质具有独特的免疫刺激能力，超出了其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，这种能力与其刺激抗原呈递细胞（APC）和增加共刺激分子CD86和II类MHC表达的能力直接相关。此外，我们发现 PorB 刺激和激活抗原呈递细胞的能力直接归因于与模式识别受体 TOLL 样受体 2 (TLR2) 的相互作用，并且需要 TLR 接头蛋白 MyD88。我们进一步表明，如初步数据部分所述，TLR2 需要与 TLR1（而不是 TLR6）异二聚化，以便 PorB 刺激这些 APC。 利用这些信息，研究 PorB 免疫刺激佐剂活性的下一个合理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 PorB 与其他佐剂相比诱导的免疫反应类型，可以更好地使用 PorB 作为佐剂。此外，我们认为有必要继续研究 PorB 的佐剂潜力，因为它是目前唯一正在研究作为疫苗佐剂的 TLR2 配体。我们将比较 PorB 与其他佐剂的免疫刺激作用，包括唯一获得许可的佐剂明矾（炎性体介导的免疫激活）、其他 TLR 配体，包括 CpG DNA (TLR9​​) 和单磷酰脂质 A (MPL)(TLR4)，以及QS-21。我们将检查这些佐剂诱导对测试抗原卵清蛋白的免疫反应的能力，我们之前已使用过该抗原，如文献中所述。初步数据部分。我们将测量诱导的抗体水平、诱导的同种型和 IgG 亚型、抗体亲和力、CD4 T 细胞反应以及诱导 APC 迁移和淋巴结特异性 T 细胞刺激的能力。保护小鼠免受分泌 OVA 的李斯特菌感染。这是一个特征明确的模型，之前已被用来证明 OVA 特异性 CD4 和 CD8 T 细胞的存在对于保护小鼠免受这种病原体的感染是必要的。被构建为分泌OVA。我们将从哈佛医学院微生物学系的 Darren Higgins 博士那里获得该菌株，他在该模型方面拥有丰富的专业知识，并将担任该提案的顾问。尽管大多数获得许可的疫苗诱导的保护作用是由于中和抗体或功能性抗体的诱导，但我们认为开始明确比较这些佐剂（包括 PorB）的能力极其重要，不仅诱导功能性抗体，而且潜在地诱导保护性 CD8 CTL 反应。 公共健康相关性：我们已经证明奈瑟菌孔蛋白 PorB 是一种有效的免疫佐剂，通过 TLR2 识别和诱导抗原呈递细胞激活发挥作用。该提案将检查 PorB 的佐剂活性，并将该活性与目前研究的其他佐剂（包括明矾、CpG DNA、MPL 和 QS-21）进行比较。我们将检查体液反应和 CD8 T 细胞反应的诱导以及疫苗预防小鼠李斯特菌感染的能力。此外，我们将通过使用与表达细胞特异性cre重组酶的小鼠交配的MyD88 floxed小鼠进一步研究PorB佐剂活性的机制。