Immunoperoxidase versus specific red cell adherence in detection of ABO(H) antigens on normal urothelium: Double-blind study

In this study we compared the sensitivity of SRCA for detecting A, B, O(H) blood group antigens on the urothelial surface of normal renal pelvis, ureter, and bladder to that of immunoperoxidase staining via the avidin-biotin complex (ABC) method. In all, forty-three mucosal specimens from 23 patients were compared. There was little difference between SRCA and immunoperoxidase for the detection of A and of B antigens. H(O) antigen was detected in 94 per cent of the blood group O patients using immunoperoxidase while only 46 per cent were detected using the SRCA method. We therefore concluded that immunoperoxidase was superior to SRCA in detecting the H(O) antigen not only in normal ureter but also in normal renal pelvis and normal bladder.     

Practical utility of specific red cell adherence test in bladder cancer

There is a highly significant correlation between specific red cell adherence (SRCA) tests for ABO(H) blood group antigens and the biologic potential of a bladder cancer in terms of both superficial tumor recurrences and the subsequent development of invasive cancer. The correlation with superficial recurrences is of limited practical value because almost one half of patients with antigen-positive tumors have superficial recurrences. Similarly, because in approximately one third of patients with SRCA-negative tumors invasive cancer does not develop, SRCA tests cannot be used to select patients for early cystectomy. Because invasive cancer proceeds to develop in less than 10 per cent of patients with high-grade, antigen-positive tumors, SRCA tests may be of use to identify patients with high-grade tumors who deserve a trial of conservative therapy. SRCA tests must be interpreted with caution in patients who have been treated previously with radiation therapy or thiotepa, in whom false positive tests may occur, and in patients with type O blood in whom false negative tests may occur.     

Prognostic value of cell surface antigens using immunoperoxidase methods in bladder carcinoma

The presence of blood group cell surface antigens in carcinoma of the bladder has been shown to be associated with a relatively benign clinical course, whereas the absence of these antigens is often indicative of tumor recurrences and invasion. In practice, however, detection of these antigens using the SRCA test in tissue sections and urine specimens has been associated with some difficulties especially for O blood group individuals. We have used immunoperoxidase methods for detection of these tumor markers in tissue sections andlor urine specimens of normal individuals and patients with carcinoma of the bladder. The results were then compared with those obtained by SRCA test. Our present study indicates that the lectin antilectin immunoperoxidase method is a more sensitive and superior test for detecting antigen H in tissue sections as well as urine andlor bladderwashing specimens. It has clearly reduced the false negative test results and predicted subsequent recurrences and invasion more accurately in patients with bladder tumors. For best results in group A and B patients, both tests probably should be performed and compared.     

Blood group antigens and bladder carcinoma: A perspective

A review of the current literature relative to the measurement of blood group antigens on bladder epithelium is presented. The determination of the presence or absence of such antigens on bladder tumors has enabled us to separate patients into two predictive groups at a time when their routine histologic patterns are identical. Group I consists of those patients with detectable blood group antigens by specific red cell adherence testing (SRCA). Regardless of the grade of their lesion, these patients have a low incidence (0–19%) of the ultimate development of invasive disease. Group II consists of those patients without demonstrable blood group antigens on their bladder carcinomas. These patients have a 60 to 93 per cent chance of invasive disease developing within five years. Treatment thus can be predicated not only on the grade and stage, but also on a prediction of future behavior. Studies mapping cystectomy specimens, as well as those studying random mucosal biopsies, have shown that when the primary tumor lacks demonstrable blood group antigens, other areas of the bladder are also SRCA negative. These findings may help explain the frequent recurrrences and ultimate development of invasive lesions in some of these patients. The role of the measurement of blood group antigens in urine cytology is reviewed, as well as those situations in which red cell adherence may be less definitive, i. e., those patients with carcinoma in situ, after radiotherapy, or thiotepa, and in those patients with blood group O. Future areas of usefulness of blood group antigens are discussed including other organs, i.e., kidney, renal pelvis, testes, and pr s,state.     

Tissue blood-group antigens and prognosis in low stage transitional cell carcinoma of the bladder

The loss of A, B or H blood-group antigens from the surface of neoplastic epithelial cells has been correlated with aggressive tumor behavior. We examined this phenomenon in low stage transitional cell carcinoma of the bladder. In an analysis of biopsy material from 37 patients the absence of these antigens on the original or recurrent tumors correlated with the subsequent development of invasive disease (stage B or greater), while the presence of antigens correlated with failure to develop invasive disease. Analysis of transitional cell surface antigens may help improve the therapy of bladder cancer.     

The prognostic value of cell surface antigens in low grade, non-invasive, transitional cell carcinoma of the bladder.

Tumors in 23 patients who presented with a low grade, non-invasive transitional cell carcinoma of the bladder were studied for blood group antigens A, B or O on the cell surface. Of 14 patients without cell surface antigens initially 13 suffered an invasive tumor subsequently and 1 had diffuse carcinoma in situ. Of 9 patients with cell surface antigens initially 8 did not have an invasive recurrence during a 5 or more-year followup and 1 did. The presence or absence of blood group cell surface antigen on a low grade, non-invasive transitional cell carcinoma of the bladder would seem to have value in predicting future recurrence with muscle invasion.     

Correlation of ABH antigenicity and urine cytology results in transitional cell carcinoma of bladder

Loss of ABH antigens from the cell surface of transitional cell carcinoma of the bladder has been proposed as an indicator of increased cellular dedifferentiation and the tendency toward invasive recurrence. An attempt was made to correlate the urinary cytology and the ABH antigen status of 57 patients with histologically confirmed bladder tumors and 28 with only dysplasia on biopsy. A review of the results of surface antigenicity studies and the preoperative urine cytologies showed no significant difference in the diagnostic sensitivity of cytology between comparable antigen negative and positive groups.     

Specific red cell adherence test in benign and malignant lesions of the prostate

The specific red cell adherence test (SRCA) for blood group antigens has been shown to have some bearing on the invasive potential of bladder tumours. Hitherto there have been few data published from patients with prostatic disease. The results of SRCA testing in 69 such patients are presented. Each of the 30 cases of adenocarcinoma was antigen negative. However, as 18 of 39 patients with only benign hyperplasia were also antigen negative, the test clearly does not reflect extant tumour and is probably not an indicator of subsequent growth of prostatic cancer. Antigen expression was also negative in sections showing prostatitis. As the test was invariably negative in patients with adenocarcinoma, whether or not metastases were present and whatever the degree of differentiation of the primary tumour, it lacks the power to discriminate invasive potential.     

Blood group isoantigens ABO (H) in transitional carcinoma of the bladder: a clinicopathological study

Blood group A, B and H antigens were investigated in 183 paraffin embedded biopsies from 58 patients with transitional cell carcinoma of the urinary bladder, by a modified specific red cell adherence test, direct immunofluorescence with Ulex Europeus lectin and indirect immunoperoxidase method with monoclonal antibodies against blood group antigens. The results were correlated with pathological grade and stage and with the clinical course of patients evaluating the recurrence index and clinical state. Histological findings were roughly correlated with the expression of red cell tissue antigens but not with the presence of precursor H substance in biopsies from patients of blood group A or B, in which a higher proportion of H positive results was appreciated. The clinical course was also related to the presence or absence of blood group antigens in referential biopsies: 90 per cent of negative biopsies corresponded to patients who had high recurrence index whereas 75 per cent of positive biopsies corresponded to patients who had low recurrence index or did not have recurrence for five years; 25 per cent of recurrences observed in patients with referential positive biopsy were invasive whereas the proportion of invasive tumor in recurrence from negative biopsies rises to 73 per cent. In addition, all the final biopsies from patients who died of bladder tumor were negative for blood group antigens. The diagnostic and prognostic significance of these tissue antigens in transitional cell carcinoma of the urinary bladder is discussed, and we conclude that the analysis of blood group antigens in bladder biopsies by established techniques is a useful tool in clinical pathology for the screening and followup of bladder tumors, as previously suggested.     

Significance of isoantigen in the management of the urinary bladder tumor--a basic study of ABO isoantigen in step sections of the entire bladder

Twenty cases of bladder cancer which had been treated by radical cystectomy were investigated, five out of these twenty were rendered to a specific red cell adherence test (SRCA test) using step section of entire bladder and for the other 15 cases malignant lesions and surrounding non-malignant lesions, which had been determined by histological mapping, were compared using the SRCA test. The SRCA test positive and negative lesions could coexist within the same tumor. Thus in the case of a large tumor, multiple biopsies were required to evaluate the results of the SRCA test. Since multiple bladder cancer lesions of the same patient revealed different results of the SRCA test, each lesion should be evaluated individually. Some dysplasia, down growth and squamous metaplasia were SRCA test negative and this phenomenon was frequently observed in the surrounding tissue of the bladder cancer. Histologically normal bladder epithelium and hyperplasia showed SRCA test positive by 100%. All the CIS showed SRCA test negative. Positive percentage of SRCA test in grade II and grade III tumors were 40% and 23.1% respectively. There was no relationship between the pathological stage and the results of SRCA test. There was no relationship between the existence of the intramural lymphatic invasion and/or the intramural venous invasion and the result of the SRCA test. From these results, we conclude that the SRCA test is a useful tool to predict the malignant potential of the bladder cancer, but when we utilize this test for the bladder biopsy specimens we found its reliability limited.     

Immunohistochemical demonstration of blood group antigens in neoplastic and normal human urothelium: a comparison with standard red cell adherence

The specific red cell adherence test as a method to detect blood group antigen deletion in urothelial malignancy has been reported to yield approximately 40 per cent false negative results in 0 blood group patients. Our study of multiple sections of 8 normal ureters from blood group 0 patients and more than 220 specimens of transitional cell cancer taken from 48 patients reveals that the immunoperoxidase technique is more specific than the specific red cell adherence method in predicting subsequent invasion in blood group O(H) patients presenting with superficial transitional cell carcinomas (71 compared to 29 per cent) but is no more specific for tumors containing A or B antigens. However, immunoperoxidase staining does improve discernment of underlying histologic detail and, thereby, facilitates recognition of false positive antigen testing associated with squamous and adenomatous metaplasia. Areas of squamous and adenomatous metaplasia in specimens we tested were frequently antigen positive in invasive tumors. Therefore, we believe that these areas must be disregarded in determining antigen deletion in transitional cell carcinomas.     

Detection of blood group surface antigens of urinary bladder tumours using monoclonal antibodies with the Avidin-Biotin complex technique

We examined 8 normal bladder transitional epithelia and 65 transitional cell carcinomas of the urinary bladder of various stages and grades for the presence of ABO(H) blood group surface isoantigens using the immunoperoxidase staining via the Avidin-Biotin Complex (ABC) methods. It was found that 27% of patients with grade I tumours, 50% with grade II tumours and 82% with grade III tumours had loss of cell surface isoantigens. When correlated with the clinical stage the tumours showed no surface isoantigens in stage D, while 65% of patients with stage A tumours were positive for surface antigens. From among 37 patients (57%), 28 (43%) survived for more than five years. Our results suggest that surface antigens of transitional cell carcinoma of the urinary bladder tended to disappear as the histologic changes of the tumour progressed. It also was noted that a loss of ABH(O) surface isoantigens was a bad prognostic sign.     

Detection of cell surface antigen in cancer of renal pelvis and ureter

Thirty-five patients with transitional cell carcinoma of the renal pelvis or ureter of all stages and grades were studied for presence or absence of ABO(H) antigens utilizing an improved technique for staining and preserving the slides. Seventy per cent of the grade I tumors retained their antigens. Patients with antigen present had a longer duration of disease-free interval. Specific red cell adherence (SRCA) may predict the clinical course of patients with low-stage, low-grade transitional cell carcinomas and may be helpful in selecting patients for optimal therapy.     

ABO(H) cell surface antigens in parathyroid adenoma and hyperplasia

Forty-three cases of primary hyperparathyroidism were studied with the specific red cell adherence test (SRCA) to determine the presence or absence of ABO(H) cell surface antigens on abnormal parathyroid tissue. Of the 27 patients with the clinicopathologic diagnosis (CPD) of adenoma, 24 (89%) had lost the ABO(H) cell surface antigen of the abnormal gland. Among the 15 patients with the CPD of hyperplasia, the parathyroid tissue from three (20%) had lost its red cell antigen. In one patient, a metastasis from a parathyroid carcinoma had lost the ABO surface antigen. Several patients in whom conflicting SRCA and CPD were obtained had factors that raised doubts as to the validity of their CPD. The SRCA is a simple test that may aid in the difficult differentiation between parathyroid adenoma and hyperplasia.     

Specific red cell adherence test applied to tumors of ureter and renal pelvis

The specific red cell adherence (SRCA) test used previously as a prognostic indicator of bladder tumors was used in a retrospective review of 14 patients with transitional cell carcinoma of the ureter and renal pelvis. SRCA-positive individuals appear to have better survival and progress less frequently to metastatic disease. Of five patients with positive reactions, none had distant metastases. Conversely, in SRCA-negative individuals metastatic disease developed more frequently (4 of 9). The SRCA test can be as useful a test of the prognosis of ureteral and renal pelvis tumors as it is of urinary bladder tumors.     

Cell surface antigen in bilharzial bladder tumours

Detection of the ABO (H) blood group in the urothelium in cases of transitional cell carcinoma of the bladder is a good indicator of the future behaviour of these tumours. Since conventional histopathological and clinical examination in bilharzial bladder carcinoma shows a clinical staging error rate of 37.2%, we used the specific red cell adherence (SRCA) technique to study the cell surface antigens in 28 patients over a 5-year period. A prediction rate of 63.6 to 84.6% was obtained when correlated with clinical behaviour.     

Antigenic deletion and prognosis of patients with stage A transitional cell bladder carcinoma.

The mixed cell agglutination reaction was studied in 22 patients with stage A transitional cell bladder tumors. The absence of antigens on the original tumor was found to correlate with development of invasive cancer on followup. The presence of antigens on the original tumor correlated with failure to develop stage B or greater disease within 5 to 14 years of followup.     

A, B, O (H) antigenic determinants in superficial transitional cell tumors of bladder

The initial tumors of 21 patients who earlier had presented with superficial noninvasive transitional cell carcinoma of the bladder and who subsequently underwent cystectomy for invasive disease were examined by the specific red cell adherence test. Eighteen of 21 initial tumors (85.7%) were antigen negative. The time from initial tumor resection ranged from twelve to 168 months (mean thirty-eight months). Loss of blood group antigens indicated enhanced biologic aggressiveness of tumors. However the interval to invasion is sufficiently variable as to preclude the use of this assay for timing of radical cystectomy.     

Segmental resection in the management of bladder carcinoma

Segmental resection for bladder carcinoma was performed on 144 patients at our medical center between 1945 and 1971. Of these 144 cases 101 had sufficient documentation to provide meaningful data for a retrospective study. The 5-year survival rates for patients with invasive bladder carcinoma, expressed as a function of the stage of disease, were stage A--79 per cent, stage B1--80 per cent, stage B2--45 per cent and stage C--6 per cent. The combined survival rates for patients with stages O, A and B1 tumors and for those with B2 and C disease were 83 and 27 per cent, respectively. A significant observation was that of the patients with stages O, A and B1 disease only 7 per cent had a history of antecedent tumor. In contrast, 59 per cent of the patients with stage B2 and C disease had antecedent tumor.     

Prognostic validity of the specific red cell adherence test in upper urothelial tumours

The specific red cell adherence (SRCA) test for blood group antigens was used in 32 patients with upper urinary tract tumours to determine whether their survival could be predicted by this test. The SRCA results correlated well with survival in each grade and stage of the tumours. Overall, patients with positive tests showed a significantly higher 5-year survival rate than those with negative tests (87.2% vs. 24.6%). These findings add support to the prognostic importance of this test in upper urinary tract tumours.