title: Preparation and characterization of chicken intraepithelial leukocytes.
abstract: Intraepithelial leukocytes (IELs) form an important component of the intestinal immune system. Several methods of preparing chicken IELs were compared for cell yield, cell populations, and functional activity against a natural killer (NK)-susceptible lymphoblastoid cell line (LSCC-RP9). In addition, an attempt was made to immortalize IELs by infection with reticuloendotheliosis virus (REV) for use in cell-trafficking studies. Intestinal fragments were incubated in 5 mM DL-dithiothreitol at 41 degrees C for 15 min followed by three 45 min incubations in 0.1 mM EDTA. Gentle shaking of the intestinal fragments every 5 min caused considerably less damage to the epithelial cell layer than slow stirring with a magnetic bar. The latter method yielded more cells, but these were not functionally active in the NK-cell assay. The more gentle method resulted in a lower cell yield, but these cells were able to lyse LSCC-RP9; the percentage of lymphocytes present was lower but these contained a higher proportion of large granular lymphocytes. Infection of IELs with REV did not immortalize these cells.
authors.affiliation: Department of Avian and Aquatic Animal Medicine, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
authors.forename: L
authors.initials: L
authors.lastname: Hoggenmueller
authors.forename: P S
authors.initials: PS
authors.lastname: Wakenell
authors.forename: K A
authors.initials: KA
authors.lastname: Schat