|Budget Amount *help
¥4,000,000 (Direct Cost : ¥4,000,000)
Fiscal Year 1990 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1989 : ¥3,400,000 (Direct Cost : ¥3,400,000)
This project is aimed to study primarily the developing the universal influenza vaccine, which is widely effective, independent of the variation of the viral hemagglutinin gene in relation to the anti-idiotypic antibody against the antibody directed to the influenza virus receptor.
During the course of this study, following new results were obtained.
1) A new simple and improved assay method to determine the receptor specificity of influenza virus variant has been developed. This method is based on the binding of the virus to the sialoglycosphingolipids (gangliosides) or sialogly-coprotein developed in silica gel thin layer plate or film blotted from the gel after electrophoresis.
2) The simple method for the fractionation and isolation of receptor gangliosides for influenza viruses was developed. This method is based on the mapping the ganglioside species by chromatography using a new anion exchange beads (Q-sepharose).
3) ALL the base sequences of the gene and the receptor specificity of
viral hemagglutinin subtype (H1 - H13) were determined. It was found that NeuAcalpha2, 3 (6) Galbeta1, 3 (6) GlcNAcbeta1- is the most active receptor determinant which is commonly recognized by all the hemagglutin subtypes. The recogn ition of the hemadgglutinin the linkage between NeuAc and Gal (alpha2, 3 and alpha2, 6) was not always depend on the amino acid 226 (Leu or Gln) in the receptor binding pocket in the hemagglutinin molecule.
4) Single amino acid substitution of Ser (205) located distant from the receptor binding pocket, antigenic site D of A/Mem/1/71-Bel/42 (H3N1), to Tyr changed the receptor binding specificity from 2, 3 to 2, 6. Ser (205) is located across the subunit interface from the receptor-binding site. The bultky hydrophobic tyr in the variant may cause a conformational change in the receptor-binding pocket on the neighboring subunit and influence the receptor binding.
5) A new monoclonal antibody (NS24) directed to sialylparagloboside (NeuAcalpha2, 3Galbeta1, 4GlcNAcbeta1, 3Galbeta1, 4Glcbeta1-Ceramide), a common receptor gangioside of influenza A viruses was prepared. This antibody was highly specific toNeuAc2, 3Gal1, 4GlcNAc1- residue of sialylparagloboside and inhibit the binding of the A virus to erythrocytes which had been treated with sialidase and then reconstituted with sialylparagloboside. This antibody may be useful as an universal influenza vaccine effective to the wide variety of subtypes of influenza virus hemagglutinin.
6) The strategy for the developing the universal influenza virus vaccine was discussed in relation to the anti-idiotypic antibody against the antibody directed to the influenza virus receptor, sialylparagloboside.
7) A new common competitive inhibitor of influenza virus neuraminidase, ganglioside which contained thio-glycoside linkage between NeuAc and Gal was found. Less