1. Viral-antigen dependence and T-cell receptor expression in leucocytes from rhabdovirus immunized trout
- Author
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A. Ezquerra, Julio Coll, Amparo Estepa, and Francisco Carantoña Álvarez
- Subjects
endocrine system ,animal structures ,Trout ,Receptors, Antigen, T-Cell, alpha-beta ,animal diseases ,Lymphocyte ,Immunology ,Kidney ,Antigen-dependent cells ,Virus ,law.invention ,Antigen ,law ,Leukocytes ,medicine ,Animals ,T-cell receptor ,Antigens, Viral ,Cells, Cultured ,General Veterinary ,biology ,urogenital system ,T-cells ,Rhabdovirus ,Flow Cytometry ,biology.organism_classification ,Virology ,Molecular biology ,body regions ,medicine.anatomical_structure ,Cell culture ,Oncorhynchus mykiss ,biology.protein ,Recombinant DNA ,Immunization ,Rabbits ,Rhabdoviridae ,Antibody ,Cell Division - Abstract
This work describes the characterization of trout haematopoietic in vitro long-term cell cultures showing specific viral antigen-dependent cell (ADC) proliferation. The ADC cultures were developed from outbreed trout after surviving viral hemorrhagic septicaemia virus (VHSV) infections or after immunization with purified VHSV. For in vitro long-term proliferation of the ADC cultures, adherent (Ad) cells obtained from autologous trout were pulsed with VHSV recombinant glycoprotein G4 (G4-pulsed Ad cells) and added periodically to the cultures. ADC did not proliferate in cultures obtained from non-infected control trout treated in parallel with G4 or from VHSV survivor/VHSV immunized trout kidney donors treated with non-viral proteins. After months in culture, the ADC acquired an increasingly homogeneous morphology compatible with that of mature trout lymphocytes, secreted supernatant `factors', and were stained with rabbit antibodies to the ectodomain of recombinant trout T-cell receptor (TcR) β-chain. Together with all the above mentioned properties, the presence of TcR sequences in the ADC cultures confirmed by the expression of α- and β-chain TcR by nested PCR amplification and sequencing of the amplified bands, suggests that these ADC cultures contain trout T-like cells engaged in a VHSV response. These trout ADC cultures offer a first opportunity to further analyze fish anti-viral immunological responses.
- Published
- 1999