Gastric parietal cell antibodies: demonstration by immunofluorescence of their reactivity with surface of the gastric parietal cells.

Viable, intact gastric cells were obtained by pronase digestion of inverted rat stomach. The cell suspensions contained two main distinct cell population, i.e. 'large' cells (mean diameter 16 microns) and 'small' cells (mean diameter 8.5 microns). By indirect immunofluorescence on smears of dispersed rat gastric cells, the large cells were identified as parietal cells, since all the sera containing parietal cell antibodies (PCA) were seen to react with the cytoplasm of these cells, leaving the cytoplasm of the small cells completely unstained. Thirty-one PCA-positive sera and forty-one PCA-negative sera were tested for gastric cell surface-reactive antibodies by an indirect immunofluorescence technique on suspensions of viable gastric cells. All the PCA-containing sera yielded a membrane immunofluorescence confined to the large cells, while none of the PCA-negative sera induced this fluorescent pattern. The surface reaction persisted unmodified when F(ab')2 fragments processed from IgG PCA-positive sera and FITC-conjugated pepsin fragments of rabbit IgG directed against the F(ab')2 fragments of human IgG were employed for the membrane fluorescence studies. The absorption of PCA-positive sera with viable, intact gastric cells led to the disappearance of both the surface immunofluorescence of the viable large cells and the cytoplasmic fluorescence of the rat parietal cells. These results demonstrate that PCA invariably react with an antigen represented on the surface of parietal cells, and that this antigen is immunologically identical to the intracytoplasmic 'microsomal' antigen.