Methodologic and genetic influence on immunohistochemical demonstration and semiquantitation of blood group antigen A in human ureter urothelium.

In order to improve the accuracy and prognostic value of ABH blood group antigen loss in urothelial tumors, the effect of Lewis blood type and methodologic factors on detectability and distribution of blood group antigen A in human formalin-fixed, paraplast-embedded urothelium and endothelium was investigated by means of the Tween 20-modified indirect immunoperoxidase staining technique. Urothelium of Lea-b+ and Lea-b- individuals expressed significant higher amounts of blood group antigen A compared to urothelium of Lea+b- individuals. The expression on endothelial cells was related to vessel type and size, but not related to Lewis types. Compared to human anti-A, monoclonal anti-A demonstrated blood group antigen A with higher sensitivity and, due to reduced background staining, higher specificity. Consequently monoclonal anti-A detected blood group antigen A in the urothelium of Lea+b- individuals where human anti-A failed to stain, and different staining patterns became apparent. Both a two- to fourfold variation in the proportion between tissue section area and volume, and the volume of anti-A applied induced minor changes in sensitivity and specificity. The monoclonal anti-A method and knowledge about erythrocyte Lewis types might prove valuable in evaluating changes in blood group antigen-A expression in urothelial tumors.

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The effect of Tween 20 on indirect immunoperoxidase staining of blood group antigen A in human urothelium.

Journal of Histochemistry & Cytochemistry ◽

10.1177/32.9.6205049 ◽

1984 ◽

Vol 32 (9) ◽

pp. 935-941 ◽

Cited By ~ 14

Author(s):

B R Juhl ◽

T Nørgaard ◽

O J Bjerrum

Keyword(s):

Blood Group ◽

Blood Group Antigen ◽

Urothelial Cell ◽

Tween 20 ◽

Immunoperoxidase Staining ◽

Tissue Sections ◽

Background Staining ◽

Antigen A ◽

Indirect Immunoperoxidase ◽

Indirect Immunoperoxidase Staining

The effect of the nonionic detergent Tween 20 on background staining, sensitivity, and specificity in the indirect immunoperoxidase staining for blood group antigen A was investigated histologically and spectrophotometrically. Pretreatment of dewaxed formalin-fixed Paraplast-embedded tissue sections from human ureters with 2% Tween 20 and dilution of the first and second layer antisera with 0.05 or 2% Tween 20 significantly reduced background staining of the urothelial cell cytoplasm, ureteral stroma, and musculature. Spectrophotometrical analysis of tissue sections from hypernephroma (rich in cytoplasm), cervix (fibrous stroma), and myometrium (musculature) underlined the histological results with a significant reduction of the maximum absorbance of Tween 20-modified indirect immunoperoxidase-stained tissue sections. Sensitivity, evaluated histologically by the endpoint titers of urothelial cell membrane staining, endothelial cell staining, and focal cytoplasmic staining of urothelial cells, was not influenced by the Tween 20 treatment. The specificity was improved as the staining was highly reduced or absent in control sections subjected to Tween 20.

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Semiquantitative immunohistochemical evaluation of H-antigen expression in human ureters of different ABO- and Lewis types.

Journal of Histochemistry & Cytochemistry ◽

10.1177/33.9.3894503 ◽

1985 ◽

Vol 33 (9) ◽

pp. 867-874 ◽

Cited By ~ 14

Author(s):

B R Juhl

Keyword(s):

Correlation Coefficients ◽

Antigen Expression ◽

Staining Technique ◽

Blood Type ◽

Tween 20 ◽

Immunoperoxidase Staining ◽

Dolichos Biflorus ◽

Ulex Europaeus ◽

Urothelial Tumors ◽

The Individual

A study was undertaken to obtain baseline information for future studies of H-antigen expression in urothelial tumors. The influence of ABO- and Lewis genes on H-antigen expression in human ureters was investigated. Thirty two ureters of different ABO- and Lewis types were stained by the Tween 20-modified immunoperoxidase staining technique using Ulex europaeus agglutinin I (32 cases) and two monoclonal anti-H antibodies (10 cases). Ureters from A and AB individuals were also stained using monoclonal anti-A and Dolichos biflorus agglutinin (14 cases). The H-antigen expression in endothelium of all cases and in urothelium of Lea-b+ and Lea-b- individuals was correlated to ABO blood type. H antigen was never detected in urothelium of Lea+b- individuals. Endothelial H-antigen expression was not influenced by the Lewis types. In A and AB individuals, urothelial and endothelial H- and A-antigen and Dolichos biflorus reactivities were mainly insignificantly related, with correlation coefficients ranging numerically from 0.10 to 0.72. The study stresses the importance of relating blood group antigen H expression in urothelial tumors to the ABO-, Lewis, and secretors types of the individual.

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Immunohistochemical demonstration and localization of Lewis a and Lewis b determinants in human urothelium.

Journal of Histochemistry & Cytochemistry ◽

10.1177/33.4.2579997 ◽

1985 ◽

Vol 33 (4) ◽

pp. 309-314 ◽

Cited By ~ 10

Author(s):

B R Juhl

Keyword(s):

Monoclonal Antibodies ◽

Antigen Expression ◽

Staining Technique ◽

Immunoperoxidase Staining ◽

Urothelial Cells ◽

Lewis Antigen ◽

Baseline Information ◽

Urothelial Tumors ◽

Indirect Immunoperoxidase ◽

Lewis A

In order to obtain baseline information about Lewis antigen expression in human urothelium in order that changes during malignant transformation can be evaluated, urothelium from eight individuals of known erythrocyte Lewis types were stained by a Tween-modified indirect immunoperoxidase staining technique using goat antibodies directed toward the Lewis a and Lewis b determinants and mouse monoclonal antibodies directed toward the Lewis a determinant in serial dilutions. To evaluate the value of the method for tissue Lewis typing, eleven ureters from individuals of unknown erythrocyte Lewis types were stained using goat antibodies. The Lewis antigens were located on the cell membranes and in the cytoplasm of urothelial cells. Goat antibodies identified Lea-b-, Lea+b+, and Lea+b- urothelium. Monoclonal antibodies identified urothelium with both low and high Lewis a antigen expression as well as urothelium with no Lewis a antigen expression. Urothelial Lewis antigen expression correlated with erythrocyte Lewis types in Lea-b+ and Lea+b- individuals. In Lea-b- individuals Lewis determinants were either not detected or were expressed similarly to Lea-b+ individuals. Urothelial Lewis typing were doubtful in two out of the eleven ureters examined. The results imply that knowledge about erythrocyte Lewis type or normal tissue Lewis antigen expression is necessary for the immunohistochemical evaluation of changes in Lewis antigen expression in urothelial tumors.

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