Hunt J C, Johnson-Paepke J, Boardway K, Gutierrez R, Hampl H, Allen R, Heynen C, Desai S, Casey J, Tribby I
Department of Human Retroviruses, Abbott Diagnostics Division, Abbott Laboratories, Abbott Park, IL 60064.
AIDS Res Hum Retroviruses. 1990 Jul;6(7):883-98. doi: 10.1089/aid.1990.6.883.
Mouse monoclonal antibodies directed against the transmembrane proteins of HIV-1 or HIV-2 provided site-directed, unambiguous discrimination between HIV-1 and HIV-2 antibody-positive sera, when employed in immunoassays as competitive probes against serum antibodies. These monoclonal antibodies mapped to epitopes outside of the well-characterized immunodominant regions (IDR) of the transmembrane proteins. The monoclonal competitive immunoassay was a superior method for discrimination compared with immunoprecipitation of metabolically radiolabeled HIV envelope glycoproteins, Western blot against viral envelope glycoproteins, or noncompetitive enzyme immunoassays employing HIV recombinant transmembrane proteins or synthetic IDR peptides as serological targets. The monoclonal competitive assay was not affected by antigenic cross reactivity or nonspecific reactivity exhibited by selected serum samples toward envelope proteins or peptides, respectively. Results of the monoclonal competitive immunoassay were supported by results of a peptide inhibition assay employing free IDR peptides in competition with IDR peptides on a solid support for binding of serum antibody. IDR peptide inhibition clearly demonstrated non-cross-reactive antigenic specificity of sera toward either the HIV-1 IDR or the HIV-2 IDR. The monoclonal competitive assay also identified samples containing antibody to both HIV-1 and HIV-2 transmembrane proteins. Analysis of these samples by IDR peptide inhibition indicated they contained two distinct, non-cross-reactive populations of antibodies, one directed to the HIV-1 IDR and the other directed to the HIV-2 IDR.
当在免疫测定中用作针对血清抗体的竞争性探针时，针对HIV-1或HIV-2跨膜蛋白的小鼠单克隆抗体能够对HIV-1和HIV-2抗体阳性血清进行定点、明确的区分。这些单克隆抗体定位在跨膜蛋白特征明确的免疫显性区域（IDR）之外的表位上。与代谢性放射性标记的HIV包膜糖蛋白的免疫沉淀、针对病毒包膜糖蛋白的蛋白质免疫印迹或使用HIV重组跨膜蛋白或合成IDR肽作为血清学靶标的非竞争性酶免疫测定相比，单克隆竞争性免疫测定是一种更优越的区分方法。单克隆竞争性测定不受所选血清样品分别对包膜蛋白或肽表现出的抗原交叉反应性或非特异性反应性的影响。单克隆竞争性免疫测定的结果得到了肽抑制测定结果的支持，该肽抑制测定使用游离IDR肽与固相支持物上的IDR肽竞争血清抗体的结合。IDR肽抑制清楚地证明了血清对HIV-1 IDR或HIV-2 IDR的非交叉反应性抗原特异性。单克隆竞争性测定还鉴定出含有针对HIV-1和HIV-2跨膜蛋白抗体的样品。通过IDR肽抑制对这些样品进行分析表明，它们含有两种不同的、非交叉反应的抗体群体，一种针对HIV-1 IDR，另一种针对HIV-2 IDR。
