SV40 immortalization of feline fibroblasts as targets for MHC-restricted cytotoxic T-cell assays
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, vol.79, no.3-4, pp.285-295, 2001 (SCI-Expanded)
- Publication Type: Article / Article
- Volume: 79 Issue: 3-4
- Publication Date: 2001
- Doi Number: 10.1016/s0165-2427(01)00272-0
- Journal Name: VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
- Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- Page Numbers: pp.285-295
- Keywords: immortal cells, feline, fibroblast, SV40, feline immunodeficiency virus, CTL, SV40-MEDIATED IMMORTALIZATION, IMMUNODEFICIENCY VIRUS, EMBRYO FIBROBLASTS, ANTIGEN, LYMPHOCYTES, RESPONSES, LINES
- Akdeniz University Affiliated: No
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 Cr-51 release CTL assays in feline immunodeficiency virus (FIV)-infected cats and in vitro stimulated allogeneic T-cell cultures. (C) 2001 Elsevier Science B.V. All rights reserved.