Janvier B, Baillou A, Archinard P, Mounier M, Mandrand B, Goudeau A, Barin F
Département de Microbiologie Médicale et Moléculaire, URA CNRS 1334, CHRU Bretonneau, Tours, France.
J Clin Microbiol. 1991 Mar;29(3):488-92. doi: 10.1128/jcm.29.3.488-492.1991.
A sequential inhibition enzyme-linked immunoassay (SIEIA) using a peroxidase-conjugated monoclonal antibody reacting to the sequence AAEWDRVHP of p24HIV-1 (amino acids 209 to 217 of p55) was developed in order to detect and determine the titer of antibody to this epitope in various populations of human immunodeficiency virus type 1 (HIV-1)-positive patients. There was a good correlation between SIEIA and a commercially available competition assay that uses recombinant p24 protein and polyclonal antibody to HIV-1 antigen, demonstrating the importance of the described epitope. Analysis of sera from French patients showed a decline of antibody to the AAEWDRVHP sequence associated with the progression of AIDS. No decrease was observed with serum samples from African patients. An immune response to the epitope was detected by SIEIA early in the course of seroconversion. Although our SIEIA uses a single p24 epitope, these data are in accordance with previously published studies in which antibodies to the whole p24 were analyzed. Sera reacting to p24 only (indeterminate profiles by Western blot [immunoblot]) did not bind to AAEWDRVHP. This epitope, which is conserved between HIV-1 and HIV-2/simian immunodeficiency virus, appears to be a major antigenic domain of p24. The area containing the sequence AAEWDRVHP and the corresponding monoclonal antibody may serve as a convenient alternative to whole purified p24 and polyclonal antibody in diagnostic and prognostic assays.
为了检测和测定1型人类免疫缺陷病毒(HIV-1)阳性患者不同群体中针对该表位的抗体滴度,开发了一种顺序抑制酶联免疫分析(SIEIA),该分析使用与p24HIV-1的AAEWDRVHP序列(p55的第209至217位氨基酸)反应的过氧化物酶偶联单克隆抗体。SIEIA与一种使用重组p24蛋白和针对HIV-1抗原的多克隆抗体的市售竞争分析之间存在良好的相关性,证明了所述表位的重要性。对法国患者血清的分析显示,与艾滋病进展相关的针对AAEWDRVHP序列的抗体下降。在非洲患者的血清样本中未观察到下降。在血清转化过程早期,通过SIEIA检测到对该表位的免疫反应。尽管我们的SIEIA使用单个p24表位,但这些数据与先前发表的分析整个p24抗体的研究一致。仅与p24反应的血清(通过蛋白质印迹法[免疫印迹]检测为不确定图谱)不与AAEWDRVHP结合。该表位在HIV-1和HIV-2/猴免疫缺陷病毒之间保守,似乎是p24的主要抗原结构域。包含序列AAEWDRVHP的区域和相应的单克隆抗体可在诊断和预后分析中作为全纯化p24和多克隆抗体的方便替代品。