摘要
The neuraminidase (NA) in viral surface is one of the main subtype-specific antigen of influenza type A viruses.Neuraminidase is an enzyme to break the bonds between hemagglutinin (HA) and sialic acid to release newly formed viruses from infected cells.In this study,the H3N2 subtype virus NA genes were sequenced and NA proteins were screened for B-cell epitopes and assessed based on immunoinformatics.Based on this results,four peptides DR6,EY7,VG8 and RE8 (covering amino acid residues 151-156,368-374,398-405 and 428-435,respectively) of the NA protein were synthesized artificially.These peptides were used to immunize New Zealand rabbits subcutaneously to raise antisera.Experimental results showed that these four peptides were capable of eliciting antibodies against H3N2 viruses in a specific and sensitive feature,detected in vitro by enzyme-linked immunosorbent assay.Moreover,hemadsorption anti-releasing effects took place in three three-antisera-mixtures at a dilution of 1:40.Alignment using NA gene database showed that amino acid residues in these four epitope peptides were substituted at specific sites in all the NAs sequenced in this study.It was suggested that these NA epitope peptides might be used in combination with HA proteins as vaccine antigens.
The neuraminidase （NA） in viral surface is one of the main subtype-specific antigen of influenza type A viruses. Neuraminidase is an enzyme to break the bonds between hemagglutinin （HA） and sialic acid to release newly formed viruses from infected cells. In this study, the H3N2 subtype virus NA genes were sequenced and NA proteins were screened for B-cell epitopes and assessed based on immunoinformatics. Based on this results, four peptides DR6, EY7, VG8 and RE8 （covering amino acid residues 151-156, 368-374, 398-405 and 428-435, respectively） of the NA protein were synthesized artificially. These peptides were used to immunize New Zealand rabbits subcutaneously to raise antisera. Experimental results showed that these four peptides were capable of eliciting antibodies against H3N2 viruses in a specific and sensitive feature, detected in vitro by enzyme-linked im- munosorbent assay. Moreover, hemadsorption anti-releasing effects took place in three three-antisera-mixtures at a dilution of 1 ： 40. Alignment using NA gene database showed that amino acid residues in these four epitope peptides were substituted at specific sites in all the NAs sequenced in this study. It was suggested that these NA epitope peptides might be used in combination with HA proteins as vaccine antigens.
基金
supported by the National Natural Science Foundation of China(30972757)
