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

进行试验以评价和表征用HIV或SIV疫苗免疫和/或感染HIV、SHIV或SIV的动物的体液免疫应答 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 测定SIV疫苗免疫或SIV感染动物的血清（或粘膜分泌物）中和疫苗所用SIV毒株对细胞系和/或原代细胞（PBMC）感染的能力。进一步表征这些抗体，包括测定针对疫苗（同源）SIV毒株的中和滴度。 测定血清（或粘膜分泌物）对SIV异源株或毒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）测定（上文）以中和SIV同源株。 培养适当的HIV、SHIV和SIV病毒储备液，测定储备液的体外滴度，并证明在使用疫苗研究血清进行中和试验之前，HIV感染者或SHIV或SIV感染猴的血清可中和病毒 进行试验，以评价和表征HIV或SIV疫苗免疫和/或HIV、SHIV或SIV感染的非人灵长类动物粘膜部位的免疫应答。 a.进行检测和测量免疫和/或感染猕猴的粘膜分泌物（如阴道和直肠拭子/洗液和唾液）中的HIV特异性或SIV特异性抗体（IgG、伊加和分泌型伊加抗体）或抗体分泌细胞的试验。包括对适当阴性对照样品的测定。 B.开发并进行试验，以评价免疫和/或感染猕猴的粘膜分泌物或粘膜组织中的功能性体液免疫应答（如上文所述的病毒中和试验）。

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.