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SV40 Immortalization of feline fibroblasts as targets for MHC-restricted cytotoxic T-cell assays.
- Source :
-
Veterinary immunology and immunopathology [Vet Immunol Immunopathol] 2001 May 30; Vol. 79 (3-4), pp. 285-95. - Publication Year :
- 2001
-
Abstract
- CTL assays in outbred cats have been difficult to perform because of a lack of a good source of syngeneic target cell. Primary fibroblasts from cats are widely used as target cells for MHC-restricted cytotoxic T-cell (CTL) assays, but their limited life-spans of 8-10 culture passages can be problematic for longitudinal studies. To circumvent the life-span limitations of primary fibroblast cultures, we developed a procedure for immortalizing feline primary fibroblast cells by transfection with a molecular clone of simian virus 40 (SV40). Fibroblast cultures from skin biopsies of 28 cats were immortalized using this procedure and have been passaged for longer than 6 months without showing any phenotypic difference from the original primary cells. Non-SV40 transfected feline fibroblasts from a selection of animals in the same group survived for only 6-8 weeks before reaching senescence. The immortalized fibroblasts expressed SV40 T-antigen and Class I MHC protein, and were successfully used as target cells in 51Cr release CTL assays in feline immunodeficiency virus (FIV)-infected cats and in vitro stimulated allogeneic T-cell cultures.
- Subjects :
- Animals
Antigens, Polyomavirus Transforming biosynthesis
Antigens, Polyomavirus Transforming genetics
Cell Transformation, Viral genetics
Cellular Senescence physiology
Chromium Radioisotopes
Cytotoxicity Tests, Immunologic methods
Epitopes, T-Lymphocyte immunology
Fibroblasts immunology
Fluorescent Antibody Technique veterinary
Histocompatibility Antigens Class I biosynthesis
Histocompatibility Antigens Class I genetics
Histocompatibility Antigens Class II biosynthesis
Histocompatibility Antigens Class II genetics
Immunodeficiency Virus, Feline isolation & purification
Isoantigens immunology
Specific Pathogen-Free Organisms
Transfection veterinary
Antigens, Polyomavirus Transforming immunology
Cats immunology
Cell Transformation, Viral immunology
Cytotoxicity Tests, Immunologic veterinary
Fibroblasts cytology
Simian virus 40 immunology
T-Lymphocytes, Cytotoxic immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0165-2427
- Volume :
- 79
- Issue :
- 3-4
- Database :
- MEDLINE
- Journal :
- Veterinary immunology and immunopathology
- Publication Type :
- Academic Journal
- Accession number :
- 11389962
- Full Text :
- https://doi.org/10.1016/s0165-2427(01)00272-0