A method for the identification of human peripheral blood T lymphocytes by sequential immunogold and esterase double staining. 1983

V Bergroth, and Y T Konttinen, and S Reitamo

A double staining method involving the sequential use of monoclonal OKT hybridoma antibodies applied in the colloidal immunogold method and followed by a simultaneously capturing azo dye method for the detection of acid alpha-naphthyl acetate esterase (ANAE) is described. Mononuclear leukocytes isolated from human peripheral blood using a Ficoll-Hypaque density gradient were stained. M-pattern ANAE-positive monocytes (diffuse staining) were excluded from the lymphocyte counts. 80 +/- 5% of all lymphocytes were T-pattern ANAE positive (dot-like staining) and 77 +/- 3% were OKT3 positive. 86 +/- 6% of all ANAE-positive T-pattern lymphocytes were also OKT3 positive, and 89 +/- 6% of all OKT3-positive lymphocytes were also ANAE positive. This indicates that ANAE is a good marker for total human T lymphocytes. 53 +/- 10% of human peripheral blood lymphocytes were OKT4 positive and 87 +/- 8% of all OKT4-positive lymphocytes were also ANAE positive. 30 +/- 6% of all lymphocytes were OKT8 positive, and only 26 +/- 18% of all OKT8-positive lymphocytes were ANAE negative. This indicates that ANAE cannot be used to distinguish T-helper and T-suppressor lymphocytes as identified by monoclonal antibodies.

UI MeSH Term Description Entries
D008722 Methods A series of steps taken in order to conduct research. Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D009283 Naphthol AS D Esterase Hydrolytic enzyme activity used as a histocytochemical test for the presence of esterases in tissue. Substrate used is 3-hydroxy-4'-nitro-2-naphthanilide chloroacetate (naphthol AS-D). ANAE,Naphthol AS D Acetatesterase,Naphthol AS D Chloroacetate Esterase,alpha-Naphthyl Acetate Esterase,alpha Naphthylesterase,alpha-Naphthylacetate Esterase,Acetate Esterase, alpha-Naphthyl,Esterase, alpha-Naphthyl Acetate,Esterase, alpha-Naphthylacetate,Naphthylesterase, alpha,alpha Naphthyl Acetate Esterase,alpha Naphthylacetate Esterase
D002265 Carboxylic Ester Hydrolases Enzymes which catalyze the hydrolysis of carboxylic acid esters with the formation of an alcohol and a carboxylic acid anion. Carboxylesterases,Ester Hydrolases, Carboxylic,Hydrolases, Carboxylic Ester
D006048 Gold Colloid, Radioactive A suspension of radioactive gold particles emitting negative beta particles and gamma irradiation. It was formerly used for liver scans and irradiation treatment of some metastatic malignancies. Radio-Colloidal Gold,Colloid, Radioactive Gold,Gold, Radio-Colloidal,Radio Colloidal Gold,Radioactive Gold Colloid
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D013194 Staining and Labeling The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts. Histological Labeling,Staining,Histological Labelings,Labeling and Staining,Labeling, Histological,Labelings, Histological,Stainings
D013601 T-Lymphocytes Lymphocytes responsible for cell-mediated immunity. Two types have been identified - cytotoxic (T-LYMPHOCYTES, CYTOTOXIC) and helper T-lymphocytes (T-LYMPHOCYTES, HELPER-INDUCER). They are formed when lymphocytes circulate through the THYMUS GLAND and differentiate to thymocytes. When exposed to an antigen, they divide rapidly and produce large numbers of new T cells sensitized to that antigen. T Cell,T Lymphocyte,T-Cells,Thymus-Dependent Lymphocytes,Cell, T,Cells, T,Lymphocyte, T,Lymphocyte, Thymus-Dependent,Lymphocytes, T,Lymphocytes, Thymus-Dependent,T Cells,T Lymphocytes,T-Cell,T-Lymphocyte,Thymus Dependent Lymphocytes,Thymus-Dependent Lymphocyte

Related Publications

V Bergroth, and Y T Konttinen, and S Reitamo
January 1979, Journal of immunological methods,
V Bergroth, and Y T Konttinen, and S Reitamo
August 1980, Clinical and experimental immunology,
V Bergroth, and Y T Konttinen, and S Reitamo
July 1983, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society,
V Bergroth, and Y T Konttinen, and S Reitamo
March 1993, Biochemical and biophysical research communications,
V Bergroth, and Y T Konttinen, and S Reitamo
January 1977, Acta dermato-venereologica,
V Bergroth, and Y T Konttinen, and S Reitamo
October 1988, Laboratory animals,
Copied contents to your clipboard!