The study of the antigenic and sialic acid binding sites on the hemagglutinin molecule of influenza virus by recombinant DNA technique.

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Full-length cDNA of HA gene derived from B-1-23 virus, a variant of A/USSR/90/77 (A/USSR/77) (HlNl) which had been selected with monoclonal antibody (mAb) 264 and contained an amino acid change at position 190 of the HAl region, was cloned into the SV40 expression vector. Hemabsorbing or fusion activities of the HA protein expressed on CV-1 cells infected with the recombinant virus were indistinguishable from those of the authentic HA protein on B-1-23 virus infected cells. The antigenicity of the expressed HA protein was examined with five monoclonal antibodies to the HA of A/USSR/77 virus. The HA protein reacted with all mAbs except for mAb264. To define the area of epitope 264, we prepared seven HA cDNAs modified by site specific mutagenesis, and cloned them into the SV40 expression vector (SVHA). Raymond et al showed that changed amino acid residues 219 (lys) and 227 (Glu) in A/Brazil/11/78, and 189 (Lys) and 225 (Asp) in A/Lackland/3/78 were presumaly included in the area of epitope 265 by the nucleotide sequence analysis. Of those amino acids, our results shows it is the residues 190 and 219 and possibly residue 189 that are involved in the epitope 264, but neither the residue 225 nor 227.