Polymorphisms of the hepatitis A virus cellular receptor 1 in African green monkey kidney cells result in antigenic variants that do not react with protective monoclonal antibody 190/4
Polymorphisms of the hepatitis A virus cellular receptor 1 in African green monkey kidney cells result in antigenic variants that do not react with protective monoclonal antibody 190/4
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DOI：
10.1128/jvi.72.7.6218-6222.1998
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发表时间：
1998-07-01
影响因子：
5.4
通讯作者：
Kaplan, GG
中科院分区：
文献类型：
--
作者：
Feigelstock, D;Thompson, P;Kaplan, GG
单克隆抗体（MAb）190/4阻断甲型肝炎病毒（HAV）与HAV细胞受体1（havcr-1）的结合，并保护非洲绿色猴肾（AGMK）克隆GL 37细胞（GL 37细胞）免受HAV感染。蛋白质印迹分析表明，两种广泛使用的AGMK细胞系BS-C-1和CV-1细胞不与MAb 190/4反应，但表达havcr-1。通过逆转录PCR从BS-C-1和CV-1细胞总RNA中扩增出编码havcr-1的cDNA。从cDNA核苷酸序列推断的氨基酸序列的比对显示，BS-C-1和CV-1 havcr-1与GL 37 havcr-1的不同之处在于在富含Cys的区域中具有两个取代，N48 H和K108 Q，以及在粘蛋白样区域中具有10至11个额外的取代加上18至22个氨基酸的插入。对GL 37 havcr-1和BS-C-1 havcr-1嵌合体的研究表明，K108 Q取代是MAb 190/4与BS-C-1和CV-1细胞缺乏反应的原因。结合研究表明，HAV与表达BS-C-1 havcr-1以及GL 37/BS-C-1 havcr-1嵌合体的狗细胞转染子结合。这些结果表明，HAVCR-1的抗原变体在AGMK细胞中表达，并且HAV与这些HAVCR-1变体的结合耐受保护性表位190/4的变化。
Monoclonal antibody (MAb) 190/4 blocks binding of hepatitis A virus (HAV) to the HAV cellular receptor 1 (havcr-1) and protects African green monkey kidney (AGMK) clone GL37 cells (GL37 cells) against HAV infection. BS-C-1 and CV-1 cells, two widely used AGMK cell lines, did not react with MAb 190/4 but expressed havcr-1, as judged by Western blot analysis, The cDNA coding for havcr-1 was amplified from BS-C-1 and CV-1 total cellular RNA by reverse transcription-PCR. Alignment of the amino acid sequences inferred from the cDNA nucleotide sequences showed that BS-C-1 and CV-1 havcr-1 differed from GL37 havcr-1 by having two substitutions in the Cys-rich region, N48H and K108Q, and 10 to 11 additional substitutions plus the insertion of 18 to 22 amino acids in the mucin-like region. Studies with chimeras of GL37 havcr-1 and BS-C-1 havcr-1 showed that the K108Q substitution was responsible for the lack of reaction of MAb 190/4 with BS-C-1 and CV-1 cells. Binding studies indicated that HAV bound to dog cell transfectants expressing the BS-C-1 havcr-1 as well as the GL37/BS-C-1 havcr-1 chimeras. These results indicate that antigenic variants of havcr-1 are expressed in AGMK cells and that binding of HAV to these havcr-1 variants tolerates changes in protective epitope 190/4.