QUANTITATIVE ANALYSIS OF BLOOD GROUP ANTIGEN OF TRANSITIONAL CELL CARCINOMA OF THE URINARY TRACT BY FLOW CYTOMETRY

We propose a new method of quantitative measurement of cell surface blood group antigen (BGA) in superificial transitional cell carcinoma (TCC) of the urinary tract by using flow cytometry. Flow cytometric analysis was performed on 22 cases of superficial urinary tract TCC registered at the department of Urology, Okayama University Hospital during April, 1992-Feb. 1993. Fresh samples were divided into two specimens. One specimen was fixed in 20% formalin and was subjected to immunohistochemical staining of BGA using avidine-biotine complex (ABC) method. The other specimen was dissociated into single cell with mincing. The single cell suspension was further divided into two specimens. One specimen was incubated on ice with anti-ABH mouse monoclonal antibody (DAKO) as a primary antibody. The other specimen was incubated on ice with mouse IgM as a negative control. These two specimens were reacted with FITC-conjugated rabbit anti-mouse IgM and propidium iodide (PI). Next, these cells were subjected to the flow cytometry using FACStar (Becton & Dickinson). By gating on the bivariate display of FSC vs. FL2 (PI), all nucleated cells were obtained separately and the positive rate was measured. The positive rate of FCM was successfully correlated with the degree of immunohistochemical stain. Compared with immunohistochemical staining thus far, this new technique provides a way to standerdize the quantitative measurement of BGA expressions.