Immunocytochemical localization of pancreatic stone protein in the human digestive tract.
Recently, in our laboratory, a protein extracted from human pancreatic stones was characterized and purified and a specific antibody was obtained. This pancreatic stone protein (PSP) was shown to have an inhibitory effect on the CaCO3 crystal growth in vitro. The cellular origin of such a protein and its repartition along the digestive tract were studied by immunolocalization (protein A-colloidal gold method) at the ultrastructural level. Surgical biopsies of pancreata from normal or chronic pancreatitis patients, needle liver biopsies, gastric mucosa, and jejunum and duodenum biopsies were minced and fixed in the Karnovsky medium or in buffered 4% paraformaldehyde. The specimens were washed in buffer, dehydrated through ethanol, and embedded in Epon 812. Ultrathin sections, collected on uncoated nickel grids, were submitted to the following reactives at room temperature: protein A 1 mg/ml, anti-PSP (1:2 to 1:100), and protein A-colloidal gold. The specificity of the localization was checked by substituting buffer or nonimmune rabbit serum to anti-PSP. The stone protein was markedly present in the zymogen granules and condensing vacuoles of the normal pancreatic acinar cells, the label was found in the acinar and ductal lumen. In chronic pancreatitis, the localization of PSP, when it occurred, was extremely weak in the acinar cells. No PSP was specifically characterized in hepatocytes, gastric mucosa, and enterocytes. However, a weak but specific reaction was found in the secretory granules of Paneth cells. These results in pancreas confirm the acinar secretory origin of the PSP and are in good agreement with its possible function in stabilizing pancreatic juice in vivo, which is normally supersaturated in calcium carbonate.(ABSTRACT TRUNCATED AT 250 WORDS)
D002135:Calcium-Binding Proteins / Q000032:analysis* / Q000302:isolation & purification; D003594:Cytoplasmic Granules / Q000032:analysis; D004792:Enzyme Precursors / Q000032:analysis; D006801:Humans; D007124:Immunoenzyme Techniques; D051847:Lithostathine; D009419:Nerve Tissue Proteins; D010179:Pancreas / Q000032:analysis*; D010189:Pancreatic Juice / Q000378:metabolism
D002135:Calcium-Binding Proteins; D004792:Enzyme Precursors; D051847:Lithostathine; D009419:Nerve Tissue Proteins; C497617:REG1A protein, human