1988 Fiscal Year Final Research Report Summary
Molecular Biologic and Molecular Epidemiologic Study of Group A Human Rotavirus
| Project/Area Number |
62570205
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Virology
|
|---|
| Research Institution | Sapporo Medical University |
|---|
Principal Investigator |
URASAWA Shozo School of Medicine, Sapporo Medical College, 医学部, 教授 (00045379)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
URASAWA Tomoko School of Allied Health Professions, Sapporo Medical College, 衛生短期大学部, 教授 (90045378)
TANIGUCHI Koki School of Medicine, Sapporo Medical College, 医学部, 講師 (40094213)
|
|---|
| Project Period (FY) |
1987 – 1988
|
| Keywords | Human rotavirus / Monoclonal antibody / Neutralization epitope / Serotype antigen / Cross reactive antigen / Viral protein / Viral RNA / 塩基配列 |
|---|
| Research Abstract |
1. By immunizing human rotavirus(HRV), we established 62 hybridomas that secrete monoclonal antibodies (Mabs) neutralizing HRV. Based on the reactivity patterns of these Mabs against various HRV strains in neutralization and enzyme-linked immunosorbent assays, they were classified into either serotype-specific or cross-reactive Mabs. Protein specificity of these mabs determined by both immunoprecipitation analysis and the use of genetically reassorted HRV strains revealed that while the majority of serotype-specific antigens resided for the most part in VP7 and partly (only a part of serotype 2-specific antigen) in VP3, cross-reactive antigens were located mostly in VP3 and partly in VP7. 2. By using these Mabs we selected antigenic mutants of KU strain (serotype 1 HRV) that resisted neutralization by the Mabs used for their selection. Cross-neutralization tests between the VP7-derected mabs and the antibody-selected antigenic mutants carried out to date identified one cross-reactive a
… More
nd five distinct serotype-specific neutralization epitopes which operationally overlapped one another and constituted a single antigenic site in VP7. The amino acid substitution in VP7 that are responsible for the antigenic alterations in the mutants were identified from sequencing of VP7 genes of the wild and mutant KU strains. All the amino acid substitutions in the antigenic mutants occurred in one of the two variable regions: amino acid 87 to 101 and 208 to 221.
Cross-neutralization tests between the VP3-directed Mabs and the corresponding antigenic mutants carried out to date identified at least three distinct cross-reactive neutralization epitopes in VP3. Sequencing of VP3 genes of the antigenic mutants also indicated three distinct neutralization epitopes in VP3: the mutants sustained a single amino acid substitution at position 305, 433 or 392 (or 439 which seemed to be located in close proximity to 392 in three dimensional structure). A synthetic peptide (amino acids 296 to 313) which included the sequence of the first neutralization epitope reacted with its corresponding Mab, suggesting that the region contains a "sequential" antigenic determinant. Less
|
