The appearance of these two drift variants was reflected in the epidemiological patterns observed in Italy in the 2002-2003 and 2004-2005 seasons; about 5,000,000 cumulative cases of influenza-like illness were reported in each of these two seasons, while during the mild epidemic season, the estimated number of cases ranged between 2.5 million and 4 million. In particular, during the early months of 2005, the Italian surveillance network observed the highest incidence in the Ն65-yearold group, since the surveillance network was established in 1999, and regional surveillances recorded lab-confirmed cases in immunized subjects as well as outbreaks in nursing homes where vaccine coverage was close to 100%, suggesting the possibility of suboptimal protection as concerns the A/H3N2 component in immunized subjects, as reported previously in other surveillances (6,10).In the present study, the antibody responses induced by the Wyoming/3/03 2004-2005 vaccine strain against the homologous variant, the 2004-2005 field isolate, and a previously circulating strain were investigated by comparing the antibody responses using hemagglutination inhibition (HI) and neutralization tests.The serum samples from vaccinees used in the present study had been obtained in the course of a previous vaccination study including 20 free-dwelling elderly individuals aged Ͼ60 years. All subjects had been vaccinated with one dose of a commercial trivalent influenza subunit vaccine containing 15 g hemagglutinin from each component. The vaccine contained a Wyoming/3/03-like reassortant as the A/H3N2 component. A blood sample was collected just before vaccination and 28 Ϯ 1 days later. HI and neutralization tests were performed, as previously described (5, 18), using three isolates collected during the 2001-2002, 2002-2003, and 2004-2005 seasons. Strains A/Genoa/47/02 (Pan-field), A/Genoa/1961/03 (Wyo-field), and A/Genoa/12/05 (Cal-field) were chosen as representative of the Panama/2007/99, Wyoming/3/03, and California/7/04 strains, respectively. The molecular characterization of the globular head region of hemagglutinin was carried out by the sequence analysis of the HA1 subunit, and the antigenic characterization of isolates was carried out by HI testing, as described elsewhere (4, 15). The phylogenetic tree that includes the isolates, vaccine, and reference strains is shown in Fig. 1, and the antigenic and molecular distances between the Wyoming/3/03 vaccine strain and Pan-field, Wyo-field, and Cal-field are shown in Table 1. The amino acid changes relative to Wyoming/3/03 observed in Pan-field and Cal-field that had a major impact on virus antigenicity are those in positions 155 and 156 (Pan-field) and in position 145 (Cal-field), which created an additional glycosylation site (6,10,14).HI and neutralization titers were transformed into binary logarithms and corrected for prevaccination state, as described by Beyer et al. (8). The observed distributions were confirmed to be normally distributed by the one-sample KolmogorovSmirnov goodness of...