H3亚型禽流感病毒血凝素特异性单克隆抗体的制备

Development of the monoclonal antibodies against hemagglutinin of H3 subtype avian influenza virus

以H3亚型禽流感病毒(AIV)免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,用血凝抑制试验(HI)筛选阳性杂交瘤细胞并克隆化。结果获得了4株针对H3亚型禽流感病毒血凝素的单克隆抗体,分别命名为1D7、1F9、5A4、5F5。这些单克隆抗体小鼠腹水HI效价为212～214。用H1、H6、H10亚型禽流感病毒各2株,H4、H5、H9和新城疫病毒(NDV)各1株进行特异性试验。结果表明:所有这些单抗仅与H3亚型AIV发生特异性HI反应,而不与其他亚型AIV及NDV反应。用28株H3亚型禽流感病毒进行排谱试验,结果证明:4株单抗均具有广谱性,其中1F9、5A4、5F5与受试的28株H3亚型AIV均反应,而1D7只与其中的26株反应。以上单抗将为控制畜禽及人类的流感提供必需的诊断试剂。

The female mice of eight-week-old were immunized with subtype H3-AIV. Spleen cells were collected and fused with SP2/0 myeloma cells after the third immunization. Hemagglutinin inhibition （HI） test was used to screen hybridoma cells and limited dilution method was performed to subclone the positive clones. After two cycles of subcloning, four McAbs against the hemagglutinin of H3-AIV were obtained, and designated as 1D7, 1F9, 5A4 and 5F5, respectively. They were proved to have high HI titers and were specific to AIV-H3, but not to react with other subtypes of AIVs and NDV. We chose 28 H3-AIV isolates to react with the four McAbs by the HI test. The McAbs of 1F9, 5A4 and 5F5 reacted with all the 28 virus strains, but 1D7 only reacted with 26 of 28. It suggested that the four McAbs can be used in epidemiological investigation of H3-AIV, and will be useful in controlling influenza of both animals and human beings.