An Escherichia coli CS31A fibrillum chimera capable of inducing memory antibodies in outbred mice following booster immunization with the entero-pathogenic coronavirus transmissible gastroenteritis virus

Autor: Elodie Rousset, Christine Martin, Michel Chavarot, Maurice Der Vartanian, Hubert Laude, Michel Contrepois, J P Girardeau
Přispěvatelé: Unité de Microbiologie (MIC), Institut National de la Recherche Agronomique (INRA), ProdInra, Migration
Rok vydání: 1997
Předmět:
[SDV]Life Sciences [q-bio]
Antibodies
Viral
medicine.disease_cause
Epitope
Mice
Coronaviridae
ComputingMilieux_MISCELLANEOUS
Coronavirus
CS31A fibrillae
Adhesins
Escherichia coli
0303 health sciences
biology
Escherichia coli Proteins
Immunogenicity
carrier-delivery system
[SDV] Life Sciences [q-bio]
Infectious Diseases
Mice
Inbred DBA
Molecular Medicine
Antibody
medicine.drug_class
Recombinant Fusion Proteins
Molecular Sequence Data
Immunization
Secondary
Monoclonal antibody
Article
Virus
Microbiology
recombinant DNA
03 medical and health sciences
Bacterial Proteins
Antigen
medicine
Animals
Amino Acid Sequence
030304 developmental biology
Antigens
Bacterial
Base Sequence
General Veterinary
General Immunology and Microbiology
030306 microbiology
Transmissible gastroenteritis virus
Public Health
Environmental and Occupational Health
biology.organism_classification
Virology
immune responses
Mice
Inbred C57BL
Haplotypes
Fimbriae
Bacterial
Mice
Inbred CBA
biology.protein
TGEV coronavirus
Immunologic Memory
Zdroj: Vaccine
Vaccine, Elsevier, 1997, 15 (2), pp.111-120
ISSN: 0264-410X
DOI: 10.1016/s0264-410x(96)00172-7
Popis: CS31A fibrillae are thin, flexible, heteropolymeric proteinaceous appendages exposed as a capsule-like material around the cell surface of certain Escherichia coli strains. Two antigenic peptides of the S spike glycoprotein (TGEV-S) amino acids (aa) 363-371 and 521-531 of the transmissible gastroenteritis virus (TGEV) were tandemly introduced in the loop-structured, variable region aa 202-218 of the major ClpG subunit protein composing the bulk of CS31A. The resulting hybrid fibrillae with a 25 aa heterologous peptide were produced at the cell surface. Using a monoclonal antibody (Mab) specific for the TGEV epitopes, purified hybrid fibrillae were analysed in Western blotting under native conditions, which showed that the two viral epitopes were recognized immunologically as an integral part of the hybrid fibrillae, and therefore that they were antigenically active. The immunogenicity of the fusion construct was evaluated with live recombinant bacteria, purified hybrid ClpG monomers, and purified chimeric CS31A polymers. Whatever the form of hybrid used as antigen, intraperitoneally immunized outbred mice elicited serum anti-TGEV peptides antibodies (Abs) with significant titres and capable of recognizing native TGEV particles, indicating that the epitopes are exposed in an immunogenic conformation in all cases. However, virus neutralization titres were only obtained after immunization with either purified polymers or monomers. Furthermore, 4 months after an ultimate immunization with 20 micrograms of hybrid fibrillae mice developed a strong anamnestic Ab response against the two TGEV peptides following booster inoculation with virions. We conclude that CS31A fibrillae carrying a combination of TGEV epitopes as insert can induce an immunological memory in outbred animals infected with TGEV, and therefore that hybrid CS31A fibrillae may prove efficient as components of a subunit vaccine.
Databáze: OpenAIRE
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