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针对HeLa细胞上作为功能性脊髓灰质炎病毒结合位点的表位产生单克隆抗体。

Production of a monoclonal antibody against an epitope on HeLa cells that is the functional poliovirus binding site.

作者信息

Nobis P, Zibirre R, Meyer G, Kühne J, Warnecke G, Koch G

出版信息

J Gen Virol. 1985 Dec;66 ( Pt 12):2563-9. doi: 10.1099/0022-1317-66-12-2563.

Abstract

Cell lines of primate origin carry receptors on their plasma membrane which are responsible for the specific binding of poliovirus. This paper describes the isolation and characterization of a monoclonal antibody reacting with the plasma membrane of HeLa cells. The antibody (D171) was selected for its protection of HeLa cells against the cytopathic effect of poliovirus type 1. This protection was found to extend to all three viral serotypes, while the replication of five other viruses in HeLa cells was not affected. The 125I-labelled purified antibody did not react with cell lines derived from pig, dog or rodents but bound specifically to all lines of human or primate origin. Immunoglobulin or Fab fragments of D171 prevented the binding of 35S-labelled poliovirus to HeLa cells. Conversely, nearly all binding sites of 125I-labelled D171 immunoglobulins or Fab fragments could be blocked after preincubation of HeLa cells with poliovirus. These results indicate that D171 recognizes the poliovirus receptor site on different susceptible cells and that practically all D171 binding sites are involved in the specific attachment of poliovirus to the plasma membrane. To determine whether the epitope recognized by D171 could be separated from the receptor for poliovirus, human-mouse cell hybrids were prepared and analysed. In all 40 clones tested, the susceptibility to poliovirus correlated with the binding of D171.

摘要

灵长类来源的细胞系在其质膜上携带负责脊髓灰质炎病毒特异性结合的受体。本文描述了一种与HeLa细胞质膜反应的单克隆抗体的分离和特性。该抗体(D171)因其能保护HeLa细胞免受1型脊髓灰质炎病毒的细胞病变效应而被选中。发现这种保护作用扩展到所有三种病毒血清型,而HeLa细胞中其他五种病毒的复制不受影响。125I标记的纯化抗体不与源自猪、狗或啮齿动物的细胞系反应,但能特异性结合所有人类或灵长类来源的细胞系。D171的免疫球蛋白或Fab片段可阻止35S标记的脊髓灰质炎病毒与HeLa细胞结合。相反,在用脊髓灰质炎病毒预孵育HeLa细胞后,几乎所有125I标记的D171免疫球蛋白或Fab片段的结合位点都可被阻断。这些结果表明,D171识别不同易感细胞上的脊髓灰质炎病毒受体位点,并且几乎所有D171结合位点都参与脊髓灰质炎病毒与质膜的特异性附着。为了确定D171识别的表位是否可以与脊髓灰质炎病毒受体分离,制备并分析了人-鼠细胞杂交体。在所有测试的40个克隆中,对脊髓灰质炎病毒的易感性与D171的结合相关。

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