Detection of MCT and MCTC types of human mast cells by immunohistochemistry using new monoclonal anti-tryptase and anti-chymase antibodies.
We developed an improved immunohistochemical technique for distinguishing human mast cells of the MCT (tryptase-positive, chymase-negative) and MCTC (tryptase-positive, chymase-positive) types utilizing a biotinylated murine anti-chymase monoclonal antibody (MAb), termed B7, and an alkaline phosphatase-conjugated murine anti-tryptase MAb, termed G3. The B7 MAb also was used to show the selective presence of chymase in mast cells. The distribution of MCT and MCTC cells in Carnoy's fluid-fixed tissue sections of human lung, skin, small intestine, and tonsils was analyzed by the new technique and the results compared to those obtained with the older method using a rabbit polyclonal antichymase antibody and a mouse anti-tryptase MAb in indirect immunoperoxidase and indirect immunoalkaline phosphatase protocols, respectively. In tissues known to contain predominantly mature mast cells, there were no quantitative differences between the two techniques, although the staining intensity achieved with the anti-chymase MAb was greater and without development of high background, compared to results achieved with the polyclonal antibody. MCT cells were the predominant type seen in the alveoli of the lung (93%) and in the small intestinal mucosa (81%). MCTC cells predominanted in the skin (99%) and in the small intestinal submucosa (77%) and, to a lesser degree, in tonsils (60%). However, in newborn foreskin tissue which contains predominantly immature forms of mast cells, 75% of all mast cells were stained uniformly and intensely with B7, whereas only 43% were stained with the polyclonal anti-chymase antibody. Therefore, the use of MAb provides for better standardization of reagents and more accurate assessment of the distribution of human MCT and MCTC cells in tissues than previously available methods.
Irani
A M
AM
Department of Pediatrics, Medical College of Virginia, Virginia Commonwealth University, Richmond.
Bradford
T R
TR
Kepley
C L
CL
Schechter
N M
NM
Schwartz
L B
LB
D000469:Alkaline Phosphatase; D000911:Antibodies, Monoclonal; D053818:Chymases; D006801:Humans; D007150:Immunohistochemistry; D007421:Intestine, Small / Q000166:cytology; D007962:Leukocytes / Q000201:enzymology; D008407:Mast Cells / Q000201:enzymology*; D014066:Palatine Tonsil / Q000166:cytology; D010447:Peptide Hydrolases / Q000032:analysis* / Q000276:immunology; D011650:Pulmonary Alveoli / Q000166:cytology; D012697:Serine Endopeptidases / Q000032:analysis* / Q000276:immunology; D012867:Skin / Q000166:cytology
D000911:Antibodies, Monoclonal; D000469:Alkaline Phosphatase; D010447:Peptide Hydrolases; C023073:tosylarginine methyl ester hydrolase; D012697:Serine Endopeptidases; D053818:Chymases