title: Generation of IL-2 dependent bovine cytotoxic T lymphocyte clones reactive against BHV-1 infected target cells: loss of genetic restriction and virus specificity.
abstract: To gain insight into the mechanisms of immunity to bovine herpesvirus type-1 (BHV-1) in particular the importance of the T-cell response, we attempted to clone bovine cytotoxic T lymphocytes that were specific for BHV-1 infected autologous target cells. A number of bovine T cell clones were generated by limiting dilution in the presence of bovine recombinant IL-2 and BHV-1 infected target cells as feeder layers. These clones were maintained in culture on crude IL-2 containing supernatants. In functional studies, 4 of the 16 T cell clones were shown to have high levels of cytotoxic activity specific for autolgous BHV-1 infected target cells with significantly lower cytotoxic activity against uninfected target cells and heterologous BHV-1 infected target cells. Continuous culturing of these 4 T cell clones, using either the crude IL-2 or high concentrations of recombinant bovine IL-2, resulted in the loss of both MHC restricted and BHV-1 specific cytotoxic activity. These clones now exhibit promiscuous type cytotoxic activity with the ability to lyse a variety of target cells. Using flow cytometric analysis, the phenotype of the T cell clones were shown to have bovine T lymphocyte characteristics including expression of the BoT8 marker. This is the first report of cloned bovine cytotoxic T lymphocytes reactive against BHV-1 and the generation from these clones of promiscuous cytotoxic activity against both virus-infected and non-infected bovine target cells.
authors.affiliation: Veterinary Infectious Disease Organization, Saskatoon, Saskatchewan, Canada.
authors.forename: M J
authors.initials: MJ
authors.lastname: Lawman
authors.forename: P
authors.initials: P
authors.lastname: Griebel
authors.forename: D L
authors.initials: DL
authors.lastname: Hutchings
authors.forename: W C
authors.initials: WC
authors.lastname: Davis
authors.forename: J
authors.initials: J
authors.lastname: Heise
authors.forename: L
authors.initials: L
authors.lastname: Qualtiere
authors.forename: L A
authors.initials: LA
authors.lastname: Babiuk
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis; D002417:Cattle; D002469:Cell Separation; D002999:Clone Cells; D003601:Cytotoxicity Tests, Immunologic; D005434:Flow Cytometry; D007242:Herpesvirus 1, Bovine / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D007376:Interleukin-2 / Q000378:metabolism*; D011994:Recombinant Proteins; D013602:T-Lymphocytes, Cytotoxic / Q000145:classification / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D007376:Interleukin-2; D011994:Recombinant Proteins