肾综合征出血热病毒血凝素单克隆抗体的建立与鉴定

用HFRSV血凝素免疫BALB/c小鼠,取脾细胞与SP2/0骨髓瘤细胞融合,共获得56株分泌抗HFRSV McAb的杂交瘤细胞系。对其中10株细胞系应用血凝抑制试验证明有9株为特异性分泌抗HFRSV血凝素McAb的杂交瘤细胞系。应用微量细胞培养中和试验证明6株具有中和活性。z株(3D_8和3G_1)具有高滴度的血凝抑制活性和中和活性。10株HFRSV血凝素McAb均能与从不同疫区,不同宿主分离的HFRSV反应,说明HFRSV血凝素具有群共同性。

Establishment and Identification of Hybridoma Cell Lines Secreting Monoclonal Antibodies against Hemagglutinin of Hemorrhagic Fevev with Renal Syndrome Virus

HFRSV haemagglutinin was harvested from suckling mice brain inoculated with HFRSV Chen's strain and prepared in high titre hemagglutinin. Fifty six hybridoma cell lines secreting McAb against HFRSV HAN were established by fusion of mouse myeloma cell line SP2/0 with spleen cells from BALB/c mice immunized with the HFRSV HAN. The fusion rate was 60% and 78%. The positive rate of holes was 15% and 44% with indirect immunofluorescence assay (IFA). The ten McAbs against HFRSVwere selected by hemagglutina-tion inhibition test ( HIT ) and neutralization test ( NT ) . The results show that Me Abs from nine lines were specially against HFRSV HAN; and that from six lines had neutralisation activity in vitro. McAbs 3D8 and 3G1 had a high HI liter and a high neutratization titer. By using cross-reactivity with IFA, the ten MeAbs showed negative IF reaction to the JEV, HSV-I and normal Vero-EG cell, antigen slides, but positive IF reactions to four HFRSV antigens ( Chen's, 81-14A, A9 and R22 strains) . It has been proved that HFRSV HAN revealed group type-common antigen to different HFRSV from patients and several rodents in various areas in China.