IMMUNIZATION OF ORAL MUCOSA FOR INDUCTION OF RECTAL AND GENITAL MOCOSAL IMMUNITY

口腔粘膜免疫以诱导直肠和生殖器粘膜免疫

• 批准号: 8357793
• 负责人: Michael K Axthelm
• 金额: $ 19.49万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357793
• 关键词: Animals; Antibodies; Antibody Formation; Antigens; B-Lymphocytes; Blood; Bronchoalveolar Lavage; CD8-Positive T-Lymphocytes; CD8B1 gene; Colon; DNA; Data; Dendritic Cells; Distal; Distant; Dose; Enzyme-Linked Immunosorbent Assay; Fossa; Frequencies; Funding; Gagging; Genital system; Grant; HIV; HIV vaccine; Immune response; Immune system; Immunity; Immunization; Infection; Influenza; Kinetics; Lingual tonsil; Lung; Macaca mulatta; Measures; Mediating; National Center for Research Resources; Nose; Oral mucous membrane structure; Organism; Peripheral; Phenotype; Plasma; Primates; Principal Investigator; Rectum; Regimen; Research; Research Infrastructure; Resources; Route; SIV; Serum; Sexually Transmitted Diseases; Site; Source; Staining method; Stains; Surface; T cell response; T-Lymphocyte; Testing; Tonsil; United States National Institutes of Health; Vaccination; Vaccines; chemokine; cost; cytokine; mucosal site; pathogen; rectal; respiratory; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Like most other sexually transmitted diseases (STDs), HIV's primary site of infection is at mucosal surfaces, and like most other STDs, there is no vaccine for HIV. The proposed study will explore the possible use of the oral mucosal route for immunization against HIV/STDs through induction of immunity at distal sites mediated via the common mucosal immune system. We will characterize innate, adaptive cellular, and humoral responses in the periphery and in mucosal (oral, nasal, pulmonary and rectal) sites to vaccines given at 2 oral mucosal sites (submucosal immunization in the buccal fossa and over the lingual tonsil). Rhesus macaques will be immunized with an SIV version of one of the leading HIV vaccine regimens: DNA prime/replication-incompetent Ad5 vaccine boost. We will test whether submucosal immunization over the lingual tonsil produces robust peripheral B and T cell responses. Adaptive responses will be measured in the blood and at mucosal sites; antigen-specific T cells will be measured using ELISpot and intracellular cytokine staining and antibody will be measured by ELISA. Possible mechanisms underlying differences in adaptive responses elicited by different routes will be investigated via studies of innate immune responses (systemic dendritic cell phenotype and function, and serum cytokine/chemokine profiles). This study will provide data on whether vaccination of the oral mucosa can provide protection against sexually transmitted organisms, through induction of immunity at distal sites (such as the rectum) mediated by the common mucosal immune system. It will also investigate the usefulness of this route of immunization for protection against respiratory pathogens, such as influenza, when immunity in the nose and lungs is desired. DNA priming elicited low to undetectable vaccine-specific cellular responses in blood. However, in bronchoalveolar lavage (BAL) high frequency polyfunctional SIV-specific CD4+and CD8+T cells were detected by ICS in DNA-primed IT animals, although DNA-primed IM animals showed very low frequency responses. CM9 gag tetramer responses were significantly higher in the IT group (P0.005). SIV-specific antibody responses in plasma were similar for IT and IM groups, but showed distinct kinetic profiles. Responses were significantly increased after boosting with rAd5; the IT group generated relatively stronger cellular responses in blood, but the difference in magnitude of responses by group was not statistically significant (%CM9+CD8+cells; IT=3.2%, IM=2.3%). High-level CD8+responses were seen at mucosal sites in the IT group (%CM9+CD8+cells: BAL=11.8%, colon=1.5%). Tonsillar immunization can elicit systemic responses and mucosal responses at distant sites. Further characterization of mucosal responses, including protection from low-dose rectal challenge with SIVmac239, is in progress.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 像大多数其他性传播疾病(STD)一样，HIV的主要感染部位是粘膜表面，而且像大多数其他STD一样，没有针对HIV的疫苗。这项拟议的研究将探索利用口腔粘膜途径通过共同的粘膜免疫系统在远端部位诱导免疫来对抗艾滋病毒/性传播疾病的可能性。我们将描述外周和粘膜(口腔、鼻腔、肺和直肠)对2个口腔粘膜部位(颊窝和舌扁桃体上的粘膜下免疫)接种疫苗的先天、适应性细胞和体液反应。恒河猴将接种一种疫苗 领先的艾滋病毒疫苗方案之一的SIV版本：DNA初始/复制无效Ad5疫苗Boost。我们将测试舌扁桃体粘膜下免疫是否产生强大的外周B细胞和T细胞反应。适应性反应将在血液和粘膜部位进行测量；抗原特异性T细胞将使用ELISpot和细胞内细胞因子染色进行测量，抗体将通过ELISA进行测量。不同途径引起的适应性反应差异的可能机制将通过研究先天性免疫反应(系统树突状细胞的表型和功能，以及血清细胞因子/趋化因子谱)来研究。这项研究将提供数据，说明口腔粘膜疫苗接种是否可以通过共同的粘膜免疫系统在远端部位(如直肠)诱导免疫，从而提供对性传播生物体的保护。它还将调查当需要鼻部和肺部免疫时，这种免疫路线在预防呼吸道病原体(如流感)方面的有用性。 DNA启动在血液中引发低到无法检测到的疫苗特异性细胞反应。然而，在DNA免疫的IT动物的支气管肺泡灌洗(BAL)中，免疫印迹法检测到高频的多功能SIV特异性的CD4+和CD8+T细胞，而DNA免疫的IM动物表现出非常低的频率反应。IT组CM9Gag四聚体反应率显著高于IT组(P0.005)。IT组和IM组血浆中SIV特异性抗体反应相似，但表现出不同的动力学特征。经rAd5增强后，免疫应答显著增加，IT组在血液中产生相对较强的细胞应答，但不同组间应答强度差异无统计学意义(%CM9+CD8+细胞；IT=3.2%，IM=2.3%)。IT组粘膜部位可见高水平的CD8+反应(%CM9+CD8+细胞：BAL=11.8%，结肠=1.5%)。扁桃体免疫可在远处引起全身反应和粘膜反应。对粘膜反应的进一步表征，包括使用SIVmac239保护其免受低剂量直肠攻击，正在进行中。