流行性出血热病毒血凝素单克隆抗体的研制与鉴定

用流行性出血热病毒(EHFV)陈株血凝素(HAN)免疫Balb/C小鼠,取其脾细胞与骨髓瘤细胞SP2/0融合,杂交瘤生长孔率为60％,阳性率为22％,三次克隆化培养阳性率达100％,通过筛选建立一株杂交瘤细胞1H8。其腹水单克隆抗体(McAb)的间接免疫荧光滴度为1:16×10~4,其血凝抑制滴度为1:5120。经染色体核型分析,1H8杂交细胞染色体为85—105条,每个细胞均含1—2条中着丝点标记染色体。经琼脂双扩散试验,1H8McAb属IgG2b亚类。该McAb与17株不同来源的EHFV都有免疫荧光反应,且对5株不同来源的EHFV—HAN具有血凝抑制活性。故1H8为血凝抑制抗体。这说明17株EHFV都具有血凝素抗原决定簇,但是1H8McAb对它们具有不同的活性。

PREPARATION AND CHARACTERIZATION OF THE MONOCLONAL ANTIBODY AGAINST THE HEMAGGLUTININ OF THE EPIDEMIC HEMORR HAGIC FEVER (EHFV)

1H_8 monocl onal antibody(McAb)was obtained by fusing SP2/0 myeloma cells with spleen cells from Balb/c mice. They were immunized with the hemagglutinin(HAN) from the Chen strain of EHFV. The success rate of growing a hybridoma cell was 60%. The positive reaction rate was 22%. The positive reaction rate of cloning reached 100% for 3 times. Finally one hybrid cell line had been established. The titers of the in the ascites McAbwere up to 1: 16x10~4 by indirect fluorescent antibody technic(IFAT) and 1: 5120 in hemagglutination inhibition test(HIT). The Karyotypes of the hybridoma cells indicated that the had 85 to 105 chromosomes, among which one or two metacentric chromosomesfrom every SP2/0 cell co uld be seen. Further more, double ,diffusion agartest 1H8 McAb was identified as mouse IgG2b subclass. In addition, it was found that 1H_8 McAb had indirect immunfluorescent reaction with 17 strains of EHFV from different sources, and had activity of HIT against 5 strains of HAN. Therefore, 1H_8 is the inhibition antibody of hemagglutination. The result implies that all the 17 strins of viruses have the antigenic(HAN) determinants, but lH_3 McAb has different activities against them.