树突状细胞对灭活口蹄疫病毒的泛素化作用

通过制备BALB/c小鼠单核细胞源树突状细胞(DC),构建了树突状细胞与灭活口蹄疫病毒(FMDV)的体外作用系统,考察树突状细胞对灭活FMDV的泛素化作用。收获荷载灭活FMDV后不同时间的树突状细胞,制备树突状细胞裂解液,用SDS-PAGE和Western-blot分析灭活FMDV抗原是否被泛素化。结果发现,在将灭活FMDV荷载于树突状细胞后的第0.5、3、6、10和20 h均可检测到分子质量约为75ku的FMDV蛋白,并且被FK2抗体标记的泛素化蛋白含量明显多于FK1抗体标记的泛素化蛋白含量。说明灭活FMDV被树突状细胞加工提呈的过程包括泛素化作用,并且泛素化蛋白主要在溶酶体内被降解。

Ubiquitination of dendritic cells on inactivated foot-and-mouth disease virus

To elucidate the mechanisms underlying the processing and presentation of the inactivated foot-and-mouth disease virus(FMDV) antigen,monocyte-derived dendritic cells(DCs) from BALB/c mouse were prepared and pulsed with inactivated FMDV in vitro to determine if ubiquitination was involved in the processing of the inactivated FMDV antigens.The ubiquitinated FMDV protein of 75 ku in molecular mass could be detected by Western-blot at 0.5,3,6,10 and 20 h immediately after FMDV were loaded onto DCs.The levels of ubiquitinated proteins recognized by FK2 were higher than those by FK1,indicating that lysosome may play a major role in the processing of FMDV antigens.The results showed that antigens of the inactivated FMDV were recognized by the ubiquitinating system and targeted to the lysosome for proces-sing in DCs.