Ts mutations in the genome of cold-adapted variants of human influenza virus A/Krasnodar/101/59 (H2N2) at different passage levels and their reproduction in lungs of hamsters.

Human influenza virus A/Krasnodar/101/59 (H2N2) was passaged in chick fibroblast cultures in the presence of trypsin at suboptimal temperature. The virus which underwent 16 passages at 28 degrees C possessed cold-adapted (ca) and temperature sensitive (ts) phenotypes and formed larger plaques at the optimal temperature (33 degrees C). Its reproduction in the lungs of hamsters was decreased as evidenced by approximately 2.5 log10 lower titres; only one of 9 virus isolates from the lungs of hamsters acquired the ts +/- phenotype, although it had retained a ca phenotype. Recombination of this variant with ts mutants of fowl plague virus (FPV) revealed a ts mutation only in gene 4 of this variant coding for haemagglutinin (HA). The virus which had had 25 passages at 28 degrees C possessed the same properties as the previous variant, but all eight virus isolates from the lungs of hamsters retained the ts phenotype; the genome of this variant contained ts mutations in genes 1, 3, 4, 5 and 6. The mutation found in gene 8 was not a ts mutation. The virus, which underwent 25 passages at 28 degrees C and additional 15 passages at 27 degrees C, formed large plaques and alike to the previous variants it possessed the ca and ts phenotypes; however, its reproduction in the lungs of hamsters was decreased by 4.0 log10 and occurred in the lungs only of 4 out 16 infected animals. This variant contained ts mutations in genes 1, 3, 4, 5, 6 and 7 and a non-ts mutation in gene 8.