Antigen presentation in the sheep: generation of antigen-specific T-cell lines

Antigen-specific sheep T-cell lines have been generated in vitro from peripheral blood mononuclear cells (PBMC). PBMC prepared from antigen-primed animals were cultured in the presence of ovalbumin (OVA) or purified protein derivative (PPD). After 5 days of culture, activated antigen-specific cells were expanded by further culture in the presence of recombinant human interleukin-2 (IL-2). Cell lines generated after two cycles of antigen stimulation followed by expansion with IL-2 show a proliferative response to antigen only in the presence of autologous antigen-presenting cells (APC) and recognize only the antigen used in the original stimulation. An OVA-specific cell line was found to be capable of recognizing a synthetic peptide corresponding to amino acid residues 323-338 of OVA. The cell lines also responded by proliferation in an allogeneic mixed leucocyte reaction (MLR). Cell-surface phenotyping shows that the cell lines comprise both CD4- and CD8-positive cells.