Morita M, Suzuki T, Nakajima K, Shiozawa C, Gill M J, Hoshino H
Division of Biomedical Research, Kitasato Institute Hospital, Tokyo, Japan.
Clin Diagn Lab Immunol. 1995 Sep;2(5):583-9. doi: 10.1128/cdli.2.5.583-589.1995.
Antibodies against reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) have been detected in seropositive subjects by immunoprecipitation, Western immunoblotting, and neutralization assay. Recently, we noticed that the antibodies against RT stabilized RT upon heat inactivation, and we have developed a stabilization assay of RT antibody. Briefly, the RT of HIV-1 is completely inactivated by incubation at 56 degrees C for 20 min, but this inactivation is inhibited in the presence of a specific antibody directed against this molecule. We examined the specificity and clinical significance of this stabilization assay. HIV-1 antibody-positive sera stabilized HIV-1 RT but not HIV-2 RT, whereas half of these sera cross-neutralized HIV-2 RT. Antibody titers against RT determined by the neutralization assay and the stabilization assay were compared with clinical characteristics. Antibodies against HIV-1 RT were much more frequently detected by the stabilization assay than by the neutralization assay. Statistically significant associations were found between stabilizing antibody titer and CD4+ cell number in peripheral blood of patients and also between antibody titer and CD4+/CD8+ ratios. These results indicate that our new stabilization assay to detect specific antibodies against RT of HIV-1 is useful as a clinical marker of infection and progress of the disease.
通过免疫沉淀、蛋白质免疫印迹法和中和试验,在血清反应阳性的受试者中检测到了针对1型人类免疫缺陷病毒(HIV-1)逆转录酶(RT)的抗体。最近,我们注意到针对RT的抗体在热灭活后能使RT稳定,并且我们开发了一种RT抗体稳定试验。简而言之,HIV-1的RT在56℃孵育20分钟后会完全失活,但在存在针对该分子的特异性抗体时,这种失活会受到抑制。我们研究了这种稳定试验的特异性和临床意义。HIV-1抗体阳性血清能稳定HIV-1 RT,但不能稳定HIV-2 RT,而这些血清中有一半能交叉中和HIV-2 RT。将通过中和试验和稳定试验测定的针对RT的抗体滴度与临床特征进行了比较。通过稳定试验检测到针对HIV-1 RT的抗体比通过中和试验更频繁。在患者外周血中,稳定抗体滴度与CD4+细胞数量之间以及抗体滴度与CD4+/CD8+比值之间发现了具有统计学意义的关联。这些结果表明,我们用于检测针对HIV-1 RT的特异性抗体的新稳定试验可作为感染和疾病进展的临床标志物。