通过免疫电子显微镜检测HIV包膜特异性抗体及其与抗体滴度和病毒中和活性的相关性。

Detection of HIV envelope specific antibodies by immunoelectron microscopy and correlation with antibody titer and virus neutralizing activity.

作者信息

Hausmann E H, Gelderblom H R, Clapham P R, Pauli G, Weiss R A

出版信息

J Virol Methods. 1987 May;16(1-2):125-37. doi: 10.1016/0166-0934(87)90037-1.

Abstract

Human antisera positive for HIV were evaluated on HTLV-IIIB producing cells by two different immunoelectron microscopic (IEM) techniques. In preembedding immunoferritin IEM a heavy label was observed with early budding HIV. Under the same conditions cell released 'mature' particles were almost negative, which could be explained by the direct observation that most of the surface glycoprotein knobs are lost spontaneously during virus maturation. Using freshly infected cultures after agarose embedding, immunogold labelling of ultrathin cryosections allowed us to detect and differentiate internal core as well as virus envelope antigens. A good qualitative correlation between neutralization titers and IEM labelling intensity was observed. This type of immunocryoultramicrotomy appears to be useful for the detection of antigens in and on the virion. It might turn out valuable for the characterization of the env gp120 epitopes of HIV.

摘要

采用两种不同的免疫电子显微镜（IEM）技术，对HIV阳性的人抗血清在产生HTLV-IIIB的细胞上进行了评估。在包埋前免疫铁蛋白IEM中，早期出芽的HIV出现了大量标记。在相同条件下，细胞释放的“成熟”颗粒几乎呈阴性，这可以通过直接观察到大多数表面糖蛋白突起在病毒成熟过程中自发丢失来解释。使用琼脂糖包埋后的新鲜感染培养物，超薄冷冻切片的免疫金标记使我们能够检测和区分内部核心以及病毒包膜抗原。观察到中和滴度与IEM标记强度之间存在良好的定性相关性。这种免疫冷冻超薄切片术似乎可用于检测病毒粒子内和表面的抗原。它可能对HIV包膜糖蛋白gp120表位的鉴定有价值。