Location of a linear epitope recognized by monoclonal antibody S16 on the hemagglutinin-esterase glycoprotein of influenza C virus.
We reported previously that monoclonal antibody S16, which had been raised against the hemagglutinin-esterase (HE) glycoprotein of influenza C/Ann Arbor/1/50 (AA/50) virus, recognizes a linear epitope present on the HE molecules of all influenza C viruses examined except for viruses belonging to a lineage represented by Aichi/1/81 (AI/81). Comparison of the deduced amino acid sequence of HE between viruses on the AI/81-related lineage and those on the others suggests that the epitope recognized by S16 is located in a region containing amino acid residue 403 and that a change from Glu to Lys at this position causes the loss of reactivity with the antibody. To prove it, the wild type (WT) HEs of AA/50 and AI/81 as well as their mutants with an amino acid substitution at residue 403 were expressed in CV-1 cells from the recombinant simian virus 40 (SV40) and tested for reactivity with S16 by immunoprecipitation. The results showed that the AA/50 virus WT and AI/81 virus mutant HEs (both having Glu at residue 403) were reactive with S16 whereas the AI/81 virus WT and AA/50 virus mutant HEs (both having Lys at residue 403) were not. Furthermore, we examined the reactivity of S16 with two synthetic peptides (corresponding to residues 397-409) that possess Glu and Lys at position 403, respectively, by enzyme-linked immunosorbent assays. The results demonstrated that the former peptide but not the latter was reactive with S16. These observations support strongly the notion described above. During this study it was also found that S16 cross-reacts with large T antigen of SV40.
Muraki
Y
Y
Department of Bacteriology, Yamagata University School of Medicine, Iida-Nishi, Japan.
Hongo
S
S
Sugawara
K
K
Matsuzaki
Y
Y
Takashita
E
E
Kitame
F
F
Nakamura
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology*; D000952:Antigens, Polyomavirus Transforming / Q000276:immunology; D019556:COS Cells; D002460:Cell Line; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D003429:Cross Reactions; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D015870:Gene Expression; D005822:Genetic Vectors; D006023:Glycoproteins / Q000276:immunology*; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology*; D009982:Gammainfluenzavirus / Q000276:immunology*; D016296:Mutagenesis; D011995:Recombination, Genetic; D013539:Simian virus 40; D014760:Viral Fusion Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000952:Antigens, Polyomavirus Transforming; D018985:Epitopes, B-Lymphocyte; D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D014760:Viral Fusion Proteins; C060814:hemagglutinin esterase