脊髓灰质炎病毒减毒株Sabin 3经型间嵌合在人胚肺二倍体细胞(KMB17)上的适应性研究  

Studies on Adaptability of Poliovirus Attenuated Strain Sabin 3 in Human Embryonic Lung Diploid Fibroblast(KMB17) through Intertypic Recombination

作　　者：胡凝珠[1] 瞿素[1] 刘国栋[1] 姬秋彦[1] 胡云章[1] 

机构地区：[1]中国医学科学院医学生物学研究所,中国协和医科大学,昆明650118

出　　处：《病毒学报》2005年第2期88-92,共5页Chinese Journal of Virology

摘　　要：为筛选Sabin3型人胚肺二倍体细胞新适应株,将原来在体外培养过程中对KMB17细胞适应较差的Sabin3型病毒与适应较好的Sabin1型病毒混合培养11代,经噬斑筛选81个克隆后,感染性滴度检测表明,大部分筛选的克隆感染性滴度较低,仅33号克隆滴度最高,达8.125LgCCID50/ml。经全基因序列分析发现,该克隆(Sabin3—33)为Sabin1型和Sabin3型的型间嵌合株,其中5’NCR为Sabin1型,其余区域与Sabin3型相同。经型别鉴定为Sabin3型,与毒力直接相关的位点5’NCR的第480位核苷酸及位于VP2的第2034位核苷酸都未发生变异,表明已初步获得一株人胚肺二倍体细胞KMB17的新适应株。To selectively obtain a new Sabin 3 strain adapted in human embryonic lung diploid cell line KMB17 through intertypic recombination, we mixed Sabin 3 and Sabin 1 to be cultured in KMB17. Then we passaged the mixture in KMB17 for 11 passages. 81 plagues were selected and the infectious titers were measured. Results showed that the infectious titers of most selected clones were lower than that of the control,only plague No. 33 (Sabin 3-33)had the highest titer of 8.125 LgCCID50/ml. By genome sequencing, we demonstrated that Sabin 3-33 was a recombinant between Sabin 1 and Sabin 3 strains in which 5'NCR came from Sabin 1 and the other regions were from Sabin 3. Furthermore, we demonstrated that Sabin 3-33 had antigenic property of Sabin 3 strain.480th nt of 5'NCR for Sabin 1 and 2 034th nt of VP2 for Sabin 3,which directly relating to virulence, showed no mutation. The results suggested that we had obtained a new Sabin 3 strain adapted in KMB17 cell.

关 键 词：二倍体细胞 滴度 人胚 减毒株 克隆 脊髓灰质炎病毒 感染性 筛选 适应性 位点 

分 类 号：Q813[生物学—生物工程] R373.22[医药卫生—病原生物学]