Fine antigenic variation within H5N1 influenza virus hemagglutinin's antigenic sites defined by yeast cell surface display

Fifteen strains of mAb specific for HA of the A/Hong Kong/482/97 (H5N1) influenza virus were generated. The HA antigenic sites of the human A/Hong Kong/482/97 (H5N1) influenza virus were defined by using yeast cell surface‐displaying system and anti‐H5 HA mAb. Evolution analysis of H5 HA identified residues that exhibit diversifying selection in the antigenic sites and demonstrated surprising differences between residue variation of H5 HA and H3 HA. A conserved neutralizing epitope in the H5 HA protein recognized by mAb H5M9 was found using viruses isolated from 1997–2006. Seven single amino acid substitutions were introduced into the HA antigenic sites, respectively, and the alteration of antigenicity was assessed. The structure obtained by homology‐modeling and molecular dynamic methods showed that a subtle substitution at residue 124 propagates throughout its nearby loop (152–159). We discuss how the structural changes caused by point mutation might explain the altered antigenicity of the HA protein. The results demonstrate the existence of immunodominant positions in the H5 HA protein, alteration of these residues might improve the immunogenicity of vaccine strains.

[1]  Marek Michalak,et al.  Quality control in the endoplasmic reticulum. , 2010, Seminars in cell & developmental biology.

[2]  D. Kapczynski,et al.  Vaccines, Vaccination, and Immunology for Avian Influenza Viruses in Poultry , 2009 .

[3]  Gavin J. D. Smith,et al.  Characterization of Avian Influenza Viruses A (H5N1) from Wild Birds, Hong Kong, 2004–2008 , 2009, Emerging infectious diseases.

[4]  Yi Guan,et al.  Multiple Sublineages of Influenza A Virus (H5N1), Vietnam, 2005−2007 , 2008, Emerging infectious diseases.

[5]  Y. Cho,et al.  A decade of yeast surface display technology: where are we now? , 2008, Combinatorial chemistry & high throughput screening.

[6]  Yu-Chieh Liao,et al.  Identifying potential immunodominant positions and predicting antigenic variants of influenza A/H3N2 viruses. , 2007, Vaccine.

[7]  R. Webster,et al.  Epitope Mapping of the Hemagglutinin Molecule of a Highly Pathogenic H5N1 Influenza Virus by Using Monoclonal Antibodies , 2007, Journal of Virology.

[8]  David J. Stevens,et al.  Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors , 2006, Nature.

[9]  K. Yuen,et al.  Serotype 1-Specific Monoclonal Antibody-Based Antigen Capture Immunoassay for Detection of Circulating Nonstructural Protein NS1: Implications for Early Diagnosis and Serotyping of Dengue Virus Infections , 2006, Journal of Clinical Microbiology.

[10]  Y. Guan,et al.  The development and characterization of H5 influenza virus vaccines derived from a 2003 human isolate. , 2006, Vaccine.

[11]  Derek J Smith,et al.  Predictability and Preparedness in Influenza Control , 2006, Science.

[12]  Y. Guan,et al.  Establishment of multiple sublineages of H5N1 influenza virus in Asia: implications for pandemic control. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[13]  W. Zhou,et al.  Comprehensive Antibody Epitope Mapping of the Nucleocapsid Protein of Severe Acute Respiratory Syndrome (SARS) Coronavirus: Insight into the Humoral Immunity of SARS , 2005, Clinical chemistry.

[14]  Hong Jin,et al.  Two residues in the hemagglutinin of A/Fujian/411/02-like influenza viruses are responsible for antigenic drift from A/Panama/2007/99. , 2005, Virology.

[15]  Min-Shi Lee,et al.  Predicting Antigenic Variants of Influenza A/H3N2 Viruses , 2004, Emerging infectious diseases.

[16]  D. Suarez,et al.  Effect of Vaccine Use in the Evolution of Mexican Lineage H5N2 Avian Influenza Virus , 2004, Journal of Virology.

[17]  A. Lapedes,et al.  Mapping the Antigenic and Genetic Evolution of Influenza Virus , 2004, Science.

[18]  V. Cheng,et al.  Sensitive and Specific Monoclonal Antibody-Based Capture Enzyme Immunoassay for Detection of Nucleocapsid Antigen in Sera from Patients with Severe Acute Respiratory Syndrome , 2004, Journal of Clinical Microbiology.

[19]  Jonathan Dushoff,et al.  Codon bias and frequency-dependent selection on the hemagglutinin epitopes of influenza A virus , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[20]  N. Ferguson,et al.  Ecological and immunological determinants of influenza evolution , 2003, Nature.

[21]  R. Webster,et al.  Structure of antigenic sites on the haemagglutinin molecule of H5 avian influenza virus and phenotypic variation of escape mutants. , 2002, The Journal of general virology.

[22]  G. Schild,et al.  Preparation of vaccines against H5N1 influenza. , 2002, Vaccine.

[23]  Y. Muraki,et al.  Antigenic structure of the haemagglutinin of human influenza A/H2N2 virus. , 2001, The Journal of general virology.

[24]  W. Fitch,et al.  Predicting the evolution of human influenza A. , 1999, Science.

[25]  R. Webster,et al.  Human influenza A H5N1 virus related to a highly pathogenic avian influenza virus , 1998, The Lancet.

[26]  V. Hinshaw,et al.  Neutralizing epitopes of the H5 hemagglutinin from a virulent avian influenza virus and their relationship to pathogenicity , 1989, Journal of virology.

[27]  J. Yewdell,et al.  The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype) , 1982, Cell.

[28]  I. Wilson,et al.  Structural identification of the antibody-binding sites of Hong Kong influenza haemagglutinin and their involvement in antigenic variation , 1981, Nature.

[29]  Gerd Ritter,et al.  Recombinant antigen expression on yeast surface (RAYS) for the detection of serological immune responses in cancer patients. , 2003, Cancer immunity.

[30]  K D Wittrup,et al.  Yeast surface display for directed evolution of protein expression, affinity, and stability. , 2000, Methods in enzymology.