title: Functional characterization of a swine CD4(+)/CD8(+) double positive lymphoblastoid T-cell line with a CD25(+)/CD45RA(-) phenotype generated in vitro with interleukin-2.
abstract: A dual expressing (CD4(+)/CD8(+)) porcine lymphoblastoid T-cell line (pIL-2d) generated from peripheral blood mononuclear (MN) cells shown to be highly responsive to exogenous interleukin-2 (IL-2) was characterized. The swine MN cells were initially stimulated with concanavalin A (Con A), and sub-passaged using decreasing amounts of conditioned medium (CM), which was prepared from culture fluids of Con A activated porcine MN cells, until a steady growth was observed. The resulting pIL-2d cells require exogenous IL-2 from CM and are highly responsive to recombinant human IL-2 (rhIL-2). The pIL-2d cells exhibited a specific, dose-dependent proliferative response to stimulation with IL-2. The specificity of this proliferative response was confirmed to be IL-2 induced by its inhibition with an anti-swine IL-2 receptor (alpha-swIL-2R) monoclonal antibody (mAb). Furthermore, the pIL-2d cells are highly responsive to exogenous IL-2 contained in culture fluids derived from antigen-driven blastogenic tests performed with lymphocytes of vaccinated swine. This property makes the pIL-2d cells an ideal functional adjunct to immunochemical or molecular tests that are commonly used to measure total porcine IL-2. Interestingly, the phenotype of the pIL-2d cells after five or more passages was shown by flow cytometric analysis to be CD4(+)/CD8(+)/CD45RA(-)/CD25(+) and to remain unchanged thereafter. Although, the mechanism of selection and maintenance of the CD4(+)/CD8(+) DP cells developed here remains unclear, our data suggest that an oligoclonal or polyclonal expansion and maintenance of cells of this phenotype was mediated by exogenous IL-2.
authors.affiliation: Department of Pathobiology, University of Connecticut, Storrs, CT 06269, USA.
authors.forename: B R
authors.initials: BR
authors.lastname: Stasny
authors.forename: S
authors.initials: S
authors.lastname: De Guise
authors.forename: G
authors.initials: G
authors.lastname: Rompato
authors.forename: A E
authors.initials: AE
authors.lastname: Garmendia
mesh_terms: D000372:Agglutination Tests / Q000662:veterinary; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood; D001681:Biological Assay / Q000662:veterinary; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002460:Cell Line; D017077:Culture Media, Conditioned; D005434:Flow Cytometry / Q000662:veterinary; D016130:Immunophenotyping; D007376:Interleukin-2 / Q000276:immunology* / Q000494:pharmacology; D017493:Leukocyte Common Antigens / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D008297:Male; D011557:Pseudorabies / Q000276:immunology; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis / Q000276:immunology; D015375:Receptors, Interleukin-2 / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology; D013552:Swine / Q000097:blood / Q000276:immunology*; D014611:Vaccination / Q000662:veterinary
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D017077:Culture Media, Conditioned; D007376:Interleukin-2; D017472:Receptors, Interleukin-1; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D017493:Leukocyte Common Antigens