Primate Core Immunology - Virol. Lab.: Humoral Imm. Lab.

灵长类动物核心免疫学 - 病毒。

• 批准号: 8328475
• 负责人: David C Montefiori
• 金额: $ 17.31万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8328475
• 关键词: AIDS Vaccines; Animals; Antibodies; Biological Assay; Cell Line; Cells; Contracts; Enzyme-Linked Immunosorbent Assay; Evaluation; HIV; HIV vaccine; Immune response; Immunoglobulin A; Immunoglobulin G; Immunoglobulin-Secreting Cells; Immunology; In Vitro; Infection; Laboratories; Macaca; Measures; Monkeys; Mucous Membrane; Peripheral Blood Mononuclear Cell; Primates; SIV; SIV Vaccines; Saliva; Sampling; Secretory Immunoglobulin A; Serum; Swab; Vaccines; Vagina; Virus; Western Blotting; mucosal site; nonhuman primate; prototype; rectal; simian human immunodeficiency virus

Perform assays to evaluate and characterize the humoral immune responses of animals immunized with HIV or SIV vaccines and/or infected with HIV, SHIV, or SIV Perform ELISA, western blot or other assays to detect the presence of SIV antibodies in sera or mucosal secretions of immunized and/or infected animals Determine the capability of sera (or mucosal secretions) from animals immunized with SIV vaccines, or infected with SIV, to neutralize infection of cell lines and/or primary cells (PBMC) by the SIV strain used for the vaccine. Further characterize these antibodies, including determining the neutralization titer against the vaccine (homologous) SIV strain. Determine the neutralization titer against infection of appropriate cell lines and/or PBMC by a heterologous strain or strains of SIV for sera (or mucosal secretions) that were determined (above) to neutralize the homologous strain of SIV. Grow appropriate HIV, SHIV, and SIV virus stocks, determine the in vitro titer of the stocks, and demonstrate that the viruses can be neutralized by sera from HIV-infected people or SHIV- or SIV-infected monkeys prior to conducting neutralization assays with the sera from the vaccine studies Perform assays to evaluate and characterize immune responses at mucosal sites in nonhuman primates immunized with HIV or SIV vaccines and/or infected with HIV, SHIV, or SIV. a. Perform assays to detect and measure HIV-specific or SIV-specific antibodies (IgG, IgA, and secretory IgA antibodies) or antibody-secreting cells in mucosal secretions such as vaginal and rectal swabs/washes and saliva of immunized and/or infected macaques. Include assays on appropriate negative control samples. b. Develop and conduct assays to evaluate functional humoral immune responses (such as virus neutralization assays as described above) in mucosal secretions or mucosal tissues from immunized and/or infected macaques.

进行检测以评估和表征接种 HIV 或 SIV 疫苗和/或感染 HIV、SHIV 或 SIV 的动物的体液免疫反应 进行 ELISA、蛋白质印迹或其他测定，以检测免疫和/或感染动物的血清或粘膜分泌物中是否存在 SIV 抗体 确定用 SIV 疫苗免疫或感染 SIV 的动物的血清（或粘膜分泌物）中和用于疫苗的 SIV 菌株对细胞系和/或原代细胞 (PBMC) 的感染的能力。进一步表征这些抗体，包括确定针对疫苗（同源）SIV 毒株的中和滴度。 确定针对血清（或粘膜分泌物）的异源菌株或 SIV 菌株感染适当细胞系和/或 PBMC 的中和滴度，所述血清（或粘膜分泌物）已确定（上文）以中和 SIV 的同源菌株。 培养适当的 HIV、SHIV 和 SIV 病毒储液，确定储液的体外效价，并证明在使用 HIV 感染者或 SHIV 或 SIV 感染猴子的血清中和病毒，然后再进行中和测定。疫苗研究血清 对接种 HIV 或 SIV 疫苗和/或感染 HIV、SHIV 或 SIV 的非人灵长类动物进行测定，以评估和表征粘膜部位的免疫反应。 一个。进行检测和测量粘膜分泌物（例如免疫和/或感染猕猴的阴道和直肠拭子/洗液以及唾液）中的 HIV 特异性或 SIV 特异性抗体（IgG、IgA 和分泌性 IgA 抗体）或抗体分泌细胞。包括对适当的阴性对照样品的测定。 b.开发并进行测定以评估免疫和/或感染的猕猴的粘膜分泌物或粘膜组织中的功能性体液免疫反应（例如如上所述的病毒中和测定）。