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 mononuclear cell infiltrates in the gastric mucosa, loss of gastric parietal and chief cells and autoantibodies to the gastric H/K ATPase. Here we describe a simple non-enzymatic method for isolating cellular infiltrates from stomachs of gastric 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 analysis by flow cytometry. An 8.3 fold increase in the total number of lymphocytes from diseased stomachs compared to normal controls was observed. Total cell numbers of CD4+ and B cells were increased 4.8 fold and 39.5 fold respectively, in diseased stomachs compared with controls. No change was observed in the CD8+ T cell population. This method will allow detailed quantitative analysis of cellular infiltrates during the development of the gastric lesion and enrichment of pathogenic T cells for analysis and cloning. This procedure may have general application for the isolation of cellular infiltrates from lesion sites of other organs.
Alderuccio
F
F
Department of Pathology and Immunology, Monash University Medical School, Melbourne, Victoria, Australia.
Toh
B H
BH
Gleeson
P A
PA
van Driel
I R
IR
D000818:Animals; D001323:Autoantibodies / Q000032:analysis; D001327:Autoimmune Diseases / Q000201:enzymology / Q000209:etiology / Q000473:pathology*; D002465:Cell Movement / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D017506:H(+)-K(+)-Exchanging ATPase / Q000276:immunology; D006984:Hypertrophy / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000201:enzymology / Q000276:immunology / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D013270:Stomach / Q000201:enzymology / Q000276:immunology* / Q000473:pathology*
D001323:Autoantibodies; D017506:H(+)-K(+)-Exchanging ATPase