Establishment of a primary cell-line from the PBMCs of rabbit C2174.
(A) Detection, by PCR and Southern blot hybridization using HVMNE (p536)-specific probe, of HVMNE DNA in the rabbit cell-line C2174 both at 6 and 12 months after culture (lane 1, C2174 after 6 months of culture; lane 2, PCR-negative control; lane 3, cell line J94356 as PCR-positive control; lane 4, C2174 after 12 months of culture). (B) Electrophoretic mobility shift assay to evaluate the Jak/STAT status of the rabbit T-cell line derived from primary culture of rabbit C2174 PBMCs. Transition to IL-2 independence of the C2174PBL cell line was associated with STAT-5 activation. On the top panel, the plus and minus signs represent the use of IL-2 pulse after overnight ligand starvation. αSTAT-5 indicates the addition to the lysate of specific αSTAT-5 antibodies in lanes 2, 4, 6, 8, and 10. Resting PBMCs (lanes 1-2) and the C2174PBL cell line cultured in the presence of IL-2 (lanes 3-6) and in the absence of IL-2 (lanes 7-10) after 8 months of culture.
