Immunoreactivity of hormonally-characterized human endocrine cells against three novel anti-human chromogranin B(B11 and B13) and chromogranin A (A11) monoclonal antibodies.
Two novel monoclonal antibodies, called B11 and B13, directed exclusively against human chromogranin B (CgB) and another antibody, A11, specific for human chromogranin A (CgA), were obtained by immunization of mice with chromaffin granules, the fusion of their splenocytes, the screening of hybridomas supernatants by ELISA and immunohistochemistry, and characterization of the antibodies by two-dimensional immunoblotting. The antibodies were used in immunohistochemical tests to investigate the distribution of CgA and CgB in hormonally-identified cells of the human endocrine system. The A11 antibody confirmed the occurrence of CgA in gut EC, ECL, gastrin, secretin and neurotensin cells, pancreatic A and PP cells, parathyroid chief cells, pituitary TSH and gonadotrope cells and adrenal medullary cells. Only a fraction of CgA-immunoreactive cells in the human gut and pancreas showed C-terminus arginine-glycinamide immunoreactivity, suggesting pancreastatin storage. Both CgB antibodies showed immunoreactivity in gastrin cells, intestinal (but not gastric) EC cells, pancreatic A and PP cells, pituitary TSH and gonadotrope cells and adrenal medullary cells. In addition, the B11 antibody stained thyroid C cells and the B13 antibody stained the Golgi area of pituitary GH cells. It is concluded that most CgB is stored in the same cells showing CgA, although some CgA-rich cells, like gastric EC and ECL cells. lacked B11 and B13 immunoreactivities and some CgA-poor cells, like human thyroid C cells, showed intense B11 immunostaining.
Buffa
R
R
Department of Human Pathology and Genetics, University of Pavia, Italy.
Gini
A
A
Pelagi
M
M
Siccardi
A G
AG
Bisiani
C
C
Zanini
A
A
Solcia
E
E
D000911:Antibodies, Monoclonal / Q000276:immunology*; D053379:Chromogranin A; D053378:Chromogranin B; D002864:Chromogranins / Q000276:immunology* / Q000378:metabolism; D004702:Endocrine Glands / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D009419:Nerve Tissue Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; C506150:CHGA protein, human; C506134:CHGB protein, human; D053379:Chromogranin A; D053378:Chromogranin B; D002864:Chromogranins; D009419:Nerve Tissue Proteins