Studies on intermembrane protein transfer and establishment of preparation method of artificial membrane vaccine

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Recentlyy it has been reported that the various membrane proteins are transferred spontaneously from living cell membranes to liposomal membranes. We adopted the phenomenon in developing liposome vaccine against influenza virus. First, we studied viral protein transfer from influenza virus-infected cells to liposomes. Next, biological activity of the liposome vaccine was examined.The membrane proteins were recovered on the liposomes in both cases : One is direct transfer of protein in the condition of coexisting of donor cells and acceptor liposomes ; the other is protein transfer from the sorting buffer to the liposomes, in which buffer virus-infected cells were incubated for 1 hour without liposomes followed by adding liposomes after removing cells. Immunoprecipitation and immunoblotting experiments showed the transfer of viral protein from influenza virus-infected CV-1 celles to liposomes.The antigenic protein-incorporated liposomes were inoculated to BALB/c mice as an artificial membrane vaccine or liposome vaccine. Immunoactivity of the liposome vaccine was almost equivalent to that of inactivated virus. B-30 MDP (6-o-(2-tetradecyl hexadecanoyl)-N-acetylmuramyl-L-aranyl-isoglutamine) as an adjuvant enhanced strongly immunoactivity.