EXAMINATION OF THE IMMUNOLOGICAL RELATIONSHIPS BETWEEN FLAVIVIRUSES USING YELLOW-FEVER VIRUS MONOCLONAL-ANTIBODIES
EXAMINATION OF THE IMMUNOLOGICAL RELATIONSHIPS BETWEEN FLAVIVIRUSES USING YELLOW-FEVER VIRUS MONOCLONAL-ANTIBODIES
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DOI：
10.1099/0022-1317-66-7-1369
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发表时间：
1985-01-01
影响因子：
3.8
通讯作者：
VARMA, MGR
中科院分区：
文献类型：
--
作者：
GOULD, EA;BUCKLEY, A;VARMA, MGR
针对黄热病（YF）病毒疫苗株制备的单克隆抗体最初通过对感染YF病毒17D株的Vero细胞进行荧光显微镜检查来鉴定。当对所有黄病毒亚组的代表株进行类似检测时，这些抗体产生了从单特异性到广泛交叉反应的多种反应；在YF病毒包膜糖蛋白中至少鉴定出5个抗原结构域。单克隆抗体可区分YF病毒疫苗株（17D、17DD、FNV）、野生型病毒以及从17D病毒库中筛选出的空斑变体。从一名黄热病患者身上分离出的一株病毒在抗原性上与巴西疫苗株17DD相似。在生物学检测中与YF病毒包膜糖蛋白发生反应的几种抗体鉴定出了54000（54K）包膜糖蛋白；通过放射免疫沉淀和聚丙烯酰胺凝胶电泳还鉴定出了YF 17D病毒感染细胞中的45K和26K多肽。在未感染细胞中未发现这些多肽。它们可能是54K蛋白的短寿命前体、翻译后裂解产物或降解产物。其他抗体与病毒感染细胞裂解物中的48K多肽发生反应。这可能是针对YF病毒所描述的非结构NV3蛋白。通过荧光显微镜观察发现，它出现在未固定的感染细胞表面，但不出现在释放的病毒粒子上。
Monoclonal antibodies prepared against vaccine strains of yellow fever (YF) virus were initially characterized by fluorescence microscopy of Vero cells infected with YF virus strain 17D. When similarly tested against representatives of all flavivirus subgroups, the antibodies produced a wide spectrum of reactions ranging from the monospecific to the broadly cross-reactive; at least 5 antigenic domains in the YF virus envelope glycoprotein were identified. Monoclonal antibodies differentiated between YF virus vaccine strains (17D, 17DD, FNV), wild-type viruses and plaque variants selected from a 17D pool. One isolate from a patient with YF was antigenically similar to the Brazilian vaccine strain 17DD. Several of the antibodies reacting with the YF viral envelope glycoprotein in biological tests identified the 54,000 (54K) envelope glycoprotein; 45K and 26K polypeptides in YF 17D virus-infected cells were also identified by radioimmunoprecipitation and polyacrylamide gel electrophoresis. Neither of these polypeptides was found in uninfected cells. They may represent shortlived precursors of the 54K protein, post-translational cleavage or breakdown products. Other antibodies reacted with a 48K polypeptide in virus-infected cell lysates. This may be the non-structural NV3 protein described for YF virus. Its appearance on the surface of unfixed infected cells, but not on released virions, was demonstrated by fluorescence microscopy.