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用于检测人类免疫缺陷病毒逆转录酶特异性抗体的稳定性测定的特征及临床意义

Characteristics and clinical significance of a stabilization assay to detect specific antibodies to reverse transcriptase of human immunodeficiency virus.

作者信息

Morita M, Suzuki T, Nakajima K, Shiozawa C, Gill M J, Hoshino H

机构信息

Division of Biomedical Research, Kitasato Institute Hospital, Tokyo, Japan.

出版信息

Clin Diagn Lab Immunol. 1995 Sep;2(5):583-9. doi: 10.1128/cdli.2.5.583-589.1995.

Abstract

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.

摘要

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。

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