摘要
用流行性出血热病毒(EHFV)李株和陈株血凝素(HAN)分别免疫Balb/C小鼠,取其脾细胞和骨髓瘤细胞SP2/O融合,杂交瘤生长孔率为97.5％和60％,阳性率各为2.5％和22％,克隆化培养阳性率达到100％。通过筛选得2株杂交瘤细胞、一株为2B_7、另一株为1H_8。其腹水单克隆抗体(McAb)滴度均达1∶16×10~4—32×10~4。1H_8的血凝抑制滴度达1∶5120。经染色体核型分析,杂交细胞染色体2B_7为80—97条,1H_8为85—105条,每个细胞均含一条中着丝点标记染色体。McAb的Ig类别和IgG亚类检测结果2B_7为IgM,1H_8为IgG_(2b)。两者对19株不同来源的EHFV具有不同的反应,1H_8McAb与19株都具有不同程度的免疫荧光反应,滴度都较高,且对5株不同来源的EHFV-HAN具有血凝抑制活性,滴度在1∶1280—5120,故1H_8为血凝抑制抗体;2B_7只对12株EHFV起反应,对7株不起反应,且没有血凝抑制活性。这说明1H_8与2B_7具有不同的特性。
Two monoclonal antibodies(McAb) were obtained by fusion of SP2/O myeloma cells with spleen cells from Balb/c mice. They were respectively immunized with the Li strain and the hemagglutinin(HAN) from the Chen strain of EHFV. Two fusions had been done. The growth rate of hybridoma was 97.5% and 60%. The positive rate was 2.5% and 22%. The positive rate of cloning all achieved 100%. Finally two hybridoma cell lines had been established. They were separately designated as 2B_7 and 1H_8. The titers of two ascites McAbs were up to 1: 16×10~4-32×10~4 by indirect fluorescent anti- body technic(IFAT). The tilers of 1H_8 ascites McAb were up to 1: 5120 by hemagglutination inhibition lest(HIT). The karyotypes of hybridoma cells indicate that 2B_7 has 80 to 97 chromosomes, 1H_8 has 85 to 105 ones,among which one or two metacontric chromosomes from the SP2/O cells could be seen. Furthermore, according to double diffusion test of agar 2B_7 McAb was identified as mouse IgM and 1H_8 was IgG2b subclass. In addition, it was found that 1H_8 McAb had indirect immunofluorescent reaction with 19 different source strains of EHFV, and had activity of HIT against 5 strains HAN. 1H_8 is the inhibition antibody of hemagglutination test, but 2B_7 is not. 2B_7 McAb had reaction only with 12 strains and had no reaction with 7 strains and did not inhibit the hemagglutination reaction. The results imply that the 19 strains of virus all have the HAN antigenic determinants, but 1H_8 McAb has different activity.
关键词
出血热病毒
单克隆抗体
杂交瘤
Epidemic Hemorrhagie Fever Virus(EHFV)
Monoclonal Antibody ( McAb )
Hemagglutinin(HAN)
Hemagglutination Inhibition Test(HIT)
