Homosubtypic and heterosubtypic antibodies against highly pathogenic avian influenza H5N1 recombinant proteins in H5N1 survivors and non-H5N1 subjects

• Published in: Virology (New York, N.Y.) Vol. 454-455; pp. 254 - 262
• Main Authors: Noisumdaeng, Pirom, Pooruk, Phisanu, Prasertsopon, Jarunee, Assanasen, Susan, Kitphati, Rungrueng, Auewarakul, Prasert, Puthavathana, Pilaipan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.04.2014
• Subjects: Adult; Antibodies, Neutralizing - blood; Antibodies, Viral - blood; avian influenza; blood serum; Child; Child, Preschool; Cross Reactions; fluorescent antibody technique; genes; Genetic Vectors; Heterosubtypic antibodies; Highly pathogenic avian influenza (HPAI) H5N1 virus; Homosubtypic antibodies; Humans; Immunofluorescence assay; Infectious Disease; Influenza A virus; Influenza A Virus, H5N1 Subtype - genetics; Influenza A Virus, H5N1 Subtype - immunology; Influenza recombinant proteins; Influenza, Human - immunology; Influenza, Human - virology; Middle Aged; pandemic; plasmids; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Recombinant vaccinia virus; Survivors; Vaccinia virus; Vaccinia virus - genetics; Viral Structural Proteins - genetics; Viral Structural Proteins - immunology; virology; viruses; Western blot assay; Recombinant vaccinia virus; Immunofluorescence assay; Heterosubtypic antibodies; Highly pathogenic avian influenza (HPAI) H5N1 virus; Influenza recombinant proteins; Homosubtypic antibodies; Western blot assay
• DOI: 10.1016/j.virol.2014.02.024
• ISSN: 0042-6822; 1096-0341

Six recombinant vaccinia viruses containing HA, NA, NP, M or NS gene insert derived from a highly pathogenic avian influenza H5N1 virus, and the recombinant vaccinia virus harboring plasmid backbone as the virus control were constructed. The recombinant proteins were characterized for their expression and subcellular locations in TK− cells. Antibodies to the five recombinant proteins were detected in all 13 sequential serum samples collected from four H5N1 survivors during four years of follow-up; and those directed to rVac-H5 HA and rVac-NA proteins were found in higher titers than those directed to the internal proteins as revealed by indirect immunofluorescence assay. Although all 28 non-H5N1 subjects had no neutralizing antibodies against H5N1 virus, they did have cross-reactive antibodies to those five recombinant proteins. A significant increase in cross-reactive antibody titer to rVac-H5 HA and rVac-NA was found in paired blood samples from patients infected with the 2009 pandemic virus. •Recombinant vaccinia viruses each carrying an H5N1 influenza gene were constructed.•Recombinant virus infected cells were used as antigens in immunofluorescence assay.•Antibodies to all test antigens were found in H5N1 survivors and non-H5N1 subjects.•Antibodies to HA and NA were high and lasted long in the H5N1 survivors.•Antibodies to HA and NA significantly increased in the 2009 pandemic patients.