A NOVEL METHOD FOR ISOLATING MONONUCLEAR-CELLS FROM THE STOMACHS OF MICE WITH EXPERIMENTAL AUTOIMMUNE GASTRITIS

Autoimmune gastritis induced in BALB/c mice by neonatal thymectomy is a CD4(+) T cell-mediated disease. The disease is characterised by mono nuclear cell infiltrates in the gastric mucosa, loss of gastric pariet al and chief cells and autoantibodies to the gastric H/K ATPase. Here we describe a simple non-enzymatic method for isolating cellular infil trates from stomachs of gastritic mice by injection of medium directly into stomach walls, causing swelling and rupture. Using this method, large numbers of viable lymphocytes were released from stomachs for an alysis by flow cytometry. An 8.3 fold increase in the total number of lymphocytes from diseased stomachs compared to normal controls was obs erved. Total cell numbers of CD4(+) and B cells were increased 4.8 fol d and 39.5 fold respectively, in diseased stomachs compared with contr ols. No change was observed in the CD8(+) T cell population. This meth od will allow detailed quantitative analysis of cellular infiltrates d uring the development of the gastric lesion and enrichment of pathogen ic T cells for analysis and cloning. This procedure may have general a pplication for the isolation of cellular infiltrates from lesion sites of other organs.