SV40 immortalization of feline fibroblasts as targets for MHC-restricted cytotoxic T-cell assays

Koksoy S., PHIPPS A., HAYES K., MATHES L.

VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, cilt.79, sa.3-4, ss.285-295, 2001 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 79 Sayı: 3-4
  • Basım Tarihi: 2001
  • Doi Numarası: 10.1016/s0165-2427(01)00272-0
  • Dergi Adı: VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.285-295
  • Anahtar Kelimeler: immortal cells, feline, fibroblast, SV40, feline immunodeficiency virus, CTL, SV40-MEDIATED IMMORTALIZATION, IMMUNODEFICIENCY VIRUS, EMBRYO FIBROBLASTS, ANTIGEN, LYMPHOCYTES, RESPONSES, LINES
  • Akdeniz Üniversitesi Adresli: Hayır

Özet

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.