Bovine gut-associated lymphoid tissue--morphologic and functional studies. I. Isolation and characterization of leukocytes from the epithelium and lamina propria of bovine small intestine.

摘要: Abstract A successful technique for the isolation of highly pure suspensions viable leukocytes from small intestine cattle is described. Procedures ranging mechanical mincing to enzymatic digestion tissues were compared. The most reliable and reproducible procedure was sequential treatment with dithiothreitol (DTT), ethylenediaminetetraacetic acid (EDTA) in calcium-magnesium-free salt solutions, collagenase. Two populations mucosal obtained intestine. One population derived within epithelium (intraepithelial leukocytes, IEL), second lamina propria (lamina LPL). At least 2 × 10 6 obtainable each square centimeter intestinal mucosa either or propria. Erythrocyte rosetting immunofluorescence characterization conventional antisera monoclonal antibodies (MAB) demonstrated that IEL predominantly T cells (60%), relatively few B present (10%), while LPL contained high numbers (28%) a reasonable percentage (45%). Both cell proliferate resposne stimulation mitogens. Addition thiol compound, 2-mercaptoethanol (2-ME) strongly augmented mitogenic response both isolates. Human recombinant interleukin-2 (hr-IL-2) presence absence additional stimuli found be able induce proliferation types. These results demonstrate functinal can isolated cattle, they maintain their responsiveness mitogens exogenous cloned IL-2.