Mazzetti P, Giannecchini S, Del Mauro D, Matteucci D, Portincasa P, Merico A, Chezzi C, Bendinelli M
Department of Biomedicine and Retrovirus Center, University of Pisa, Pisa, Italy.
J Virol. 1999 Jan;73(1):1-10. doi: 10.1128/JVI.73.1.1-10.1999.
The feline immunodeficiency virus (FIV) cat model is extensively used to investigate possible vaccination approaches against AIDS in humans. Although consistent levels of protection have been achieved with FIV, as with other model systems, by immunizing with whole inactivated virus or fixed infected cells, the mechanisms responsible for protection are elusive. In previous studies we showed that cats immunized with a vaccine consisting of fixed infected cells were protected or unprotected against cell-free or cell-associated FIV challenge depending on the time interval between completion of vaccination and challenge. In an attempt to define possible humoral immune correlates of protection, selected sera harvested at the times of challenge from such cats were examined for anti-FIV-antibody titers and properties by using binding and functional immunological assays. Binding assays included quantitative Western blotting, enzyme-linked tests for antibodies to FIV glycoproteins and immunodominant linear epitopes, and tests for measuring conformation dependence and avidity of anti-viral-envelope antibodies. Functional assays included virus neutralization performed with two different cell substrates, complement- and antibody-dependent virolysis, blocking of reverse transcriptase, and an assay that measured the ability of sera to prevent FIV growth in cocultures of infected and uninfected cells. Despite the wide spectrum of parameters investigated, no correlation between vaccine-induced protection and the humoral parameters measured was noted.
猫免疫缺陷病毒(FIV)猫模型被广泛用于研究针对人类艾滋病的可能疫苗接种方法。尽管通过用全灭活病毒或固定感染细胞进行免疫,FIV与其他模型系统一样实现了一致的保护水平,但负责保护的机制仍不清楚。在先前的研究中,我们表明,用由固定感染细胞组成的疫苗免疫的猫,根据疫苗接种完成与攻击之间的时间间隔,对无细胞或细胞相关的FIV攻击有保护或无保护作用。为了确定可能的体液免疫保护相关性,通过使用结合和功能性免疫测定法,检查了在此类猫攻击时采集的选定血清的抗FIV抗体滴度和特性。结合测定包括定量蛋白质印迹、针对FIV糖蛋白和免疫显性线性表位的抗体的酶联试验,以及用于测量抗病毒包膜抗体的构象依赖性和亲和力的试验。功能测定包括用两种不同细胞底物进行的病毒中和、补体和抗体依赖性病毒溶解、逆转录酶阻断,以及测量血清在感染和未感染细胞共培养物中阻止FIV生长能力的试验。尽管研究了广泛的参数,但未发现疫苗诱导的保护与所测量的体液参数之间存在相关性。