Production of monoclonal antibody against histamine and its application to immunohistochemical study in the stomach.
A monoclonal antibody against histamine has been produced. A histamine-haemocyanin conjugate prepared using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide as a coupling agent was used for immunizing mice. Immunized mice were sacrificed to prepare monoclonal antibody using a hybridoma technique. On immunospot assay, the hybridoma culture supernatant containing a monoclonal antibody was capable of detecting 50 pmol of histamine. Using this antibody, we examined the cellular localization of histamine-like immunoreactivity in the stomach of normal or alpha-fluoromethylhistidine-treated rats and mice. Immunoreactive cells were abundant in the gastric mucosal layer. These positive cells were often located in the basal half of the fundic gland but were rare in the pyloric gland. The cells, small or medium in size, spindle or cone in shape, were intermingled with immunonegative epithelial cells. In the cytoplasm of the positive cells, granular reaction products were densely deposited. In addition, a few positive cells, identified as mast cells by Toluidine Blue staining, were distributed mainly in the submucosal and muscular layer. The antibody preabsorbed with 10 mM histamine gave no positive immunostaining. For pharmacological study, some rats were injected six times with c-fluoromethylhistidine every 8 h. In these rats, positive cells except mast cells were no longer detected. In conclusion, the monoclonal antibody produced appears to be highly specific for histamine. Its application in immunohistochemistry should provide a powerful tool for analysing the roles of histamine in enterochromaffin-like or mast cells in the stomach.
Doi
T
T
Division of Neuroanatomy, Institute of Molecular Neurobiology, Shiga University of Medical Science, Otsu, Japan.
Terai
K
K
Tooyama
I
I
Sakata
T
T
Kimura
H
H
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000378:metabolism*; D000918:Antibody Specificity; D019861:Enterochromaffin-like Cells / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005753:Gastric Mucosa / Q000187:drug effects / Q000378:metabolism*; D006632:Histamine / Q000276:immunology* / Q000378:metabolism*; D006640:Histidine Decarboxylase / Q000037:antagonists & inhibitors; D007150:Immunohistochemistry; D008297:Male; D008762:Methylhistidines / Q000494:pharmacology; D051379:Mice; D008813:Mice, Inbred ICR; D051381:Rats; D017208:Rats, Wistar; D012701:Serotonin / Q000378:metabolism; D013270:Stomach / Q000187:drug effects
D000906:Antibodies; D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D008762:Methylhistidines; D012701:Serotonin; C028309:alpha-fluoromethylhistidine; D006632:Histamine; D006640:Histidine Decarboxylase