SUMMARYB/Singapore/222/79-1ike influenza viruses isolated from three patients during the winter of 1981 to 1982 and cultured in either embryonated hens' eggs or MDCK cells were studied. Sequence analysis indicated that the haemagglutinin (HA) genes of the six virus preparations contained at least four distinct HA1 sequences which differed by up to six amino acids. Only one pair of viruses had amino acid differences between the egg-and MDCK cell-derived viral subpopulations and this change did not affect a glycosylation site. Mice infected with previously described recombinant vaccinia viruses expressing either the egg-or MDCK cell-derived HA of B/England/222/82 developed neutralizing antibodies against all of the 1982 type B viruses and were protected against intranasal challenge with these viruses. Therefore, in this model system, the minor sequence variation between the HAs of egg-and MDCK cell-derived influenza B/England/222/82 virus had no detectable effect on the induction of crossprotection.In order to compare the immune response induced by influenza virus haemagglutinins (HAs) differing by a single amino acid, we cloned the genes for a pair of such HA variants and expressed them in vaccinia virus (Rota et al., 1987). These variant HA genes were previously described as being present in either egg-derived or MDCK cell-derived subpopulations of a single isolate of influenza B/England/222/82 virus (B/Eng) (Schild et al., 1983). The single amino acid difference between the HAs was found at residue 198, which is near the receptor-binding site on the distal tip of the HA1 portion of the molecule. This change resulted in the loss of a specific N-linked glycosylation site that was present on the HA of virus from the MDCK cellderived subpopulation (Robertson et al., 1985). Our previous work has shown that mice vaccinated with the recombinant vaccinia viruses expressing either of the variant HAs were equivalently protected from infection after challenge by each of the B/Eng virus subpopulations (Rota et al