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J Gen Virol 89 (2008), 1970-1977; DOI 10.1099/vir.0.83679-0

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Detection of four genetic subgroup-specific antibodies to human metapneumovirus attachment (G) protein in human serum

Rika Endo1, Takashi Ebihara1, Nobuhisa Ishiguro1, Shinobu Teramoto1, Tadashi Ariga1, Chikako Sakata2, Akio Hayashi2, Hiroaki Ishiko2 and Hideaki Kikuta3

1 Department of Pediatrics, Hokkaido University Graduate School of Medicine, Sapporo, Japan
2 Mitsubishi Chemical Medience Corporation, Tokyo, Japan
3 Pediatric Clinic, Touei Hospital, Sapporo, Japan

Correspondence
Hideaki Kikuta
hide-ki{at}touei.or.jp

Human metapneumovirus (hMPV) strains are classified into two genetic groups, A and B, each of which is further divided in two genetic subgroups, A1, A2, B1 and B2. hMPV encodes two major surface glycoproteins, the fusion (F) and attachment (G) proteins, which may be immunogenic and protective antigens. Although the amino acid sequences of hMPV F protein are highly conserved, those of the G protein are highly variable with low amino acid identity between the two groups. To address the antigenic variation between the genetic subgroups, we developed an immunofluorescence assay (IFA) method using Trichoplusia ni (Tn5) insect cells infected with each recombinant baculovirus-expressed hMPV G (Bac-G) protein of the four genetic subgroups. The titre of each antibody to the four Bac-G proteins was measured by the IFA in 12 paired serum samples obtained from children infected with hMPV of each genetic subgroup. Although 11 of the 12 acute-phase serum samples in paired samples were negative for the antibody to any Bac-G proteins, all of the convalescent-phase serum samples in those paired samples were positive for the antibody to only one of the four Bac-G proteins of the infecting genotype of hMPV. Since the antibody response to hMPV G protein was transient and genetic subgroup-specific without cross-reactivity, four genetic subgroups on the basis of hMPV G protein could be identified as different serotypes. This assay may be useful for the study of immune responses of humans to different hMPV strains, especially for clarifying the risk of reinfection with hMPV.

A table showing the sequences of the primers used in this study is available with the online version of this paper.







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