Lucey D R, VanCott T C, Loomis L D, Bethke F R, Hendrix C W, Melcher G P, Redfield R R, Birx D L
Department of Retroviral Research, Walter Reed Army Institute of Research, Rockville, Maryland.
J Acquir Immune Defic Syndr (1988). 1993 Sep;6(9):994-1001.
Antibody to the human immunodeficiency virus (HIV)-1 principal neutralizing determinant (V3 loop) was measured by peptide enzyme-linked immunosorbent assay (ELISA) in cerebrospinal fluid (CSF) and paired serum samples of 21 HIV-seropositive patients. These patients had normal neurologic examinations and were without neurologic symptoms. Peptide ELISA demonstrated intrathecal antibody synthesis against the V3 loop of HIVMN, the V3 loop of HIVNY5, the V3 loop of HIVLAI, and the entire recombinant HIV-1MN gp120 in 21 of 21, 10 of 21, one of 21, and 12 of 21 patients, respectively. Biospecific interaction analysis (BIAcore), which requires only small amounts of CSF, was also used to detect anti-V3 CSF antibody. Fine mapping of linear epitopes within the V3 region was successful in three of five patients by Geysen PIN (PEPSCAN) ELISA and discordance between epitope specificity of CSF and serum antibody was found. While detection of CSF antibody against the V3 loop of HIVMN by peptide ELISA has been recently reported, we add to this finding using the peptide ELISA, PEPSCAN and BIAcore methodologies as well as measuring intrathecal antibody synthesis against V3 loops from HIV strains. Application of these techniques to future studies of anti-V3 antibody in CSF from persons receiving anti-HIV-1 immunizations may provide insight into the immunoregulation of the virus in the nervous system.
通过肽酶联免疫吸附测定法(ELISA)检测了21例HIV血清反应阳性患者脑脊液(CSF)及配对血清样本中针对人类免疫缺陷病毒(HIV)-1主要中和决定簇(V3环)的抗体。这些患者神经系统检查正常且无神经症状。肽ELISA显示,21例患者中有21例、10例、1例和12例分别针对HIVMN的V3环、HIVNY5的V3环、HIVLAI的V3环以及完整的重组HIV-1MN gp120存在鞘内抗体合成。生物特异性相互作用分析(BIAcore)仅需少量脑脊液,也用于检测抗V3脑脊液抗体。通过Geysen PIN(PEPSCAN)ELISA成功对5例患者中的3例进行了V3区域内线性表位的精细定位,且发现脑脊液和血清抗体的表位特异性存在不一致。虽然最近已有通过肽ELISA检测脑脊液中针对HIVMN的V3环抗体的报道,但我们在此基础上增加了使用肽ELISA、PEPSCAN和BIAcore方法,以及测量针对HIV毒株V3环的鞘内抗体合成。将这些技术应用于未来接受抗HIV-1免疫接种者脑脊液中抗V3抗体的研究,可能会为病毒在神经系统中的免疫调节提供见解。