[Influence of antibody-mediated immune pressure on neuraminidase gene mutations of avian influenza virus H9N2].

LG1 strain of avian influenza virus H9N2 was passaged continuously for 40 generations in chicken embryos with anti-LG1 maternal antibodies in 4 parallel experiments, of which 3 experiments had a stable mutation of "G" to "A" at #99 of the neuraminidase gene(NA)from the 20th passage resulting in a change of Met to Ile and 2 had a stable mutation of "A" to "G" at #473 of the NA gene from the 30th passage resulting in a change of Asn to Ser which occurred in the 50th passage of another experiment. Eighty continuous passages in chicken embryos without antibody did not have the same mutation, indicating that the mutations of the 2 positions were associated with selective pressure of antibodies. Analysis of the ratios of nonsynonium (NS) vs synonium (S) mutations of nucleic acids demonstrated that NS/S of 4 parallel experiments with antibodies was 4.6 (32/7) compared with 2.0 (16/8) of the 2 experiments without antibodies and this significant difference implied the selective pressure of antibodies.

[Mutations of the hemagglutinin gene of H9N2 subtype avian influenza viruses under selective pressure of antibody].

OBJECTIVE: To understand mutations of avian influenza virus (AIV) under the antibody selective pressures. METHODS: We continuously passed an H9N2-AIV strain LG1 in embryos with or without maternal antibody to LG1 in 6 separate serials. At the 10th, 20th, 30th, 40th and 50th passages in each serial, H9 hemagglutinatin gene (HA) sequences were determined and compared to the original LG1 strain. RESULTS: Only unstable random mutations happened in 29 sites with nonsynonymous vs synonymous (N/S) mutation ratio of 1.42 during 50 passages in 2 serials without maternal antibodies. However, multiple stable nonsynonymous mutations were detected gradually during 50 passages of 4 serials in embryos with maternal antibody to LG1 strain, and the NS/S ratio was as high as 3.46 among 45 mutated sites. CONCLUSION: The results suggested that the antibody selective pressure influenced mutations of H9N2 AIV HA gene during the passages in embryos with maternal antibody. Embryos with maternal antibody to certain viruses could be used as experimental models to study the immune selective pressures on viral mutations.