Specific Red Cell Adherence Test Research Articles

Histo-Blood Group Antigens in Oral Cancer and Potentially Malignant Disorders.

Background:Early detection of oral cancer is of critical importance because survival rates markedly improve when oral lesions are identified at an early stage. Aim of the present study is to investigate the expression of ABO (H) antigens in tissue specimens of oral cancer and potentially malignant disorders and to determine the role of ABO (H) antigens in tumour staging. Materials and Methods:A prospective study was conducted on 60 cases of oral cancer and potentially malignant diseases. Specific red cell adherence test (SRCA-test) was used for studying A, B and O (H) antigens in tissue specimens and iso-antigenicity of epithelium was graded according to degree of adherence of indicator red blood cells. Results:Among OSMF group, grade II adherence was seen in 53.3% cases, grade III in 33.3% cases, grade IV in 13.3% cases. In leukoplakia group, grade II adherence was seen in 26.7% cases, grade III adherence in 53.3% cases, grade IV adherence in 20% cases. Within the leukoplakia group, cases with dysplasia showed decreased adherence, compared with cases without dysplasia. Oral cancer group, negative adherence was seen in 13.3% cases, grade I adherence in 46.7% cases, grade II in 40% cases. In oral cancer group, antigen reactivity was less in poorly and moderately differentiated carcinoma, compared to well differentiated carcinoma. Conclusions:Antigen adherence and degree of loss of ABO (H) antigens in tissue specimens can be used for staging of the tumour.

A, B and H isoantigens in oral lesions

Expression of blood group A, B and H iso-antigens in oral mucosae was demonstrated by specific red cell adherence test in forty seven mucosal biopsies from oral cavity (14 normal, 10 leucoplakia and 23 squamous cell carcinomas). All normal mucosae gave a positive reaction whereas in malignancy only 6 cases of well differentiated squamous cell carcinomas were either weekly positive or showed a patchy reaction. Remaining cases were negative irrespective of their grading. In leucoplakia though the test was positive in all the cases, the intensity of adherence was less as compared to normal mucosa. The intensity of adherence was found to be directly related to the degree of cellular differentiation. Study of A, B and H isoantigens might help in deciding the prognosis in leukoplakia and/or early detection of malignancy.

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.

Immunohistological detection of T antigen and ABH blood group antigens in upper urinary tract tumours.

The status of T antigen was assessed by means of peanut agglutinin (PNA) used in an immunoperoxidase technique, and the status of ABH blood group antigens was determined by the specific red cell adherence (SRCA) test. Thirty-eight patients with upper urinary tract tumours were examined. Our data indicated that T antigen status did not correlate with conventional prognostic indicators such as grade and stage of tumour and survival of patients, while the expression or deletion of ABH blood group antigens did correlate well. It was concluded that ABH blood group antigen determination might provide a useful prognostic probe in upper urinary tract tumours should it find clinical application.

Peroxidase Antiperoxidase Versus Specific Red Cell Adherence in Detection of O (H) Antigen in Bladder Cancer: A Blind Study

We have previously demonstrated that the presence or absence of ABO(H) cell surface antigens correlated with stage, grade, and prognosis of patients with bladder cancer. However, because of the weak antigenecity of 0(H), certain investigators have reported a high rate of false-negative results utilizing specific red cell adherence (SRCA) test of Davidsohn. Since approximately 45% of the population is blood group 0(H), this high rate of false-negative result renders the test unreliable in an ordinary laboratory. The present study is comparing the results of SRCA to that of peroxidase antiperoxidase (PAP) technique in a double-blind fashion in 29 patients with bladder cancer having 0(H) blood group. In terms of grade, stage, frequency of recurrent tumor, and quality of slides, the results of PAP is superior over SRCA. Furthermore, the convenience, permanence, and ready availability of PAP will enhance the utility of this test in a conventional histopathologic laboratory with a high rate of accuracy and reproducibility.

Further studies on specificity of red cell adherence test: Nonmalignant bladder lesions

Specific red cell adherence test for blood group antigens was utilized in 32 nonmalignant bladder lesions, none of which was associated with bladder cancer, to determine the specificity of this test. All of the 14 lesions of cystitis cystica, cystitis glandularis, and chronic cystitis retained their antigens. Of the 18 lesions of squamous metaplasia, 13 (72 %) were antigen positive. Testing for blood group antigens showed an overall 84 per cent specific rate in 27 of the 32 nonmalignant bladder lesions.

Cell surface blood group antigens in prostatic carcinoma.

Surface blood group antigens are present to some degree in most epithelia. These antigens frequently are lost during neoplastic transformation. The authors looked for the presence or absence of surface blood group antigens in 52 cases of prostatic carcinoma of various histologic grades using the specific red blood cell adherence test. The normal prostatic tissue showed a 2+ reaction in patients with type A or B blood and 0-1+ in type O. The hyperplastic areas were 4+ for red blood cell adherence in patients with type A or B blood and 0-4+ in type O. In contrast, all malignant foci were negative for blood group antigens, no matter what the histologic grade or blood type.

ABO(H) Isoantigens in Bladder Tumors: A New Technique of Quantitative Analysis

The specific red cell adherence test appears promising to predict which superficial bladder tumor(s) ultimately will become invasive. However, presently the test cannot be quantitated and, thus, cannot be standardized. We describe a technique for the quantitative analysis of ABO(H) isoantigens in bladder tumors, which should help to standardize the test for prospective clinical trials.

The Value of Histologic Grading and Staging, Random Biopsies, Tumor and Bladder Mucosa Blood Group Antigens, in Predicting Progression of Superficial Bladder Cancer

We followed 67 patients with superficial bladder cancer (pTA, pT1) for 12-48 months (mean, 26.8 months) after surgical removal of their tumors. In addition to tumor grade and stage, we recorded morphologic characteristics of randomized mucosal biopsies and blood group antigens of the and 'normal' bladder mucosa by means of the specific red cell adherence test. Results were correlated with tumor recurrence and tumor progression of recurrent tumors. It is recommended that such measurements be used to decide on more aggressive surgical therapy or extensive (possibly lifelong) prophylactic instillation therapy, e.g. with mitomycin. Our own experience with mitomycin has shown that it can control tumor recurrence, tumor progression, and survival in such high-risk patients.

A, B and O(H) isoantigens in tumor of the urinary bladder. Preliminary results, relating the specific red cell adherence test to pathological grade and stage.

In a prospective study we have included the specific red cell adherence (SRCA) test for the identification of blood group isoantigens in the investigation of patients with bladder tumor. The preliminary results, relating the SRCA test to grade and stage in 50 patients are presented. None of the SRCA-positive tumors were of high-grade malignancy (WHO III), and only SRCA-negative tumors had invasive properties at the time of investigation. 8 of 9 patients with carcinoma in situ were SRCA-negative.

Altered cell surface antigen expression in bladder carcinoma detected by a new hemagglutinating monoclonal antibody.

A hemagglutinating monoclonal IgM antibody (MoAb145) was produced against a high incidence red blood cell membrane antigen. By the specific red cell adherence test, the antibody also reacted with human bladder epithelium; in addition, expression of the MoAb145 antigen was lost in some cases of transitional cell carcinoma of the bladder, in a manner similar to the ABH blood group. Hemagglutination studies with a panel of erythrocytes lacking specific high incidence red blood cell membrane antigens indicated that MoAb145 did not recognize ABH specificity but rather a determinant absent from rare MN variant erythrocytes, including En(a-) erythrocytes, which lack glycophorin-alpha. Failure of MoAb145 to stain, by indirect immunofluorescence, the erythroleukemia cell line K562, which expresses glycophorin-alpha and the MN blood group, and failure to inhibit MoAb145 hemagglutination with an erythrocyte sialoglycoprotein fraction that contained MN blood group activity suggests that MoAb145 does not recognize either glycophorin-alpha or the MN blood group, but rather another membrane determinant, which is altered in En(a-) erythrocytes. This study demonstrates a new epitope detected by MoAb145 that is shared between human erythrocyte membranes and bladder epithelia, and is affected by neoplastic transformation in transitional cell carcinoma of the bladder.

A Comparison of Clinical Course with Blood Group Antigen Testing by Specific Red Cell Adherence and Immunoperoxidase in Ureteral and Renal Pelvic Tumors

Specimens of transitional cell carcinoma of the ureter and renal pelvis from 20 patients were tested for blood group antigenicity using immunoperoxidase and specific reel cell adherence methods. The results of antigen testing were correlated with tumor stage and grade as well as the subsequent clinical course of the patients. The specific red cell adherence test was negative in 80 per cent and the immunoperoxidase test was negative in 40 per cent of all tumors. Of the 4 patients with positive specific red cell adherence tests 3 had high grade (II to III), invasive tumors as did 7 of 12 with tumors that were positive by immunoperoxidase testing. Blood group antigen testing did not prove helpful in predicting the clinical course of our patients. In addition, a careful review of previously published data does not support the conclusion that blood group antigen testing is a valuable predictor of upper tract tumor aggressiveness.

Mechanisms of action of intravesical treatment

Blood group isoantigens can be demonstrated immune-histologically on normal urothelium of the urinary bladder by means of the specific red cell adherence test. In preneoplastic and reactive changes of the urothelium and in dysplasia and carcinoma in situ, there is a loss of these antigens. Ultrastructural investigations of carcinoma in situ demonstrate a loss of normal membrane structures such as tight junctions, desmosomes, and glycocalyx. Of 18 patients with carcinoma in situ of the urinary bladder, 12 achieved tumor remission after topical chemotherapy with doxorubicin hydrochloride. A recurrence of blood group isoantigens was demonstrated in patients who achieved tumor remission. The antigenicity reappears before complete normalization of the urothelium. Ultrastructural investigations demonstrated normalization of cellular membrane structures in the respective biopsies. According to these findings and the current literature, the mechanisms of action of doxorubicin hydrochloride in carcinoma in situ may be due to retardation of cell cycle, cell loss by desquamation, cell death, and decrease of growth fraction.

ABO (H) cell surface antigens in benign and malignant parotid neoplasms.

Twenty-two inflammatory, benign, or malignant parotid lesions were studied by means of the specific red cell adherence test (SRCA), a modification of the Coombs' mixed cell agglutination reaction. In 22 normal parotid tissues the acinar structures were devoid of red cell agglutination, but it was present in ductal epithelium. Findings were similar in 2 cases of parotitis, 11 benign mixed tumors, and 1 malignant mixed tumor. All lacked red cell agglutination in areas of neoplastic change. Benign Warthin's tumors (4 cases) demonstrated antigenicity in the columnar epithelial component of the tumor, but lacked red cell agglutination in areas of the lymphoid component. One malignant Warthin's tumor showed agglutination in areas of normal columnar epithelium but not in areas of malignant dedifferentiation. Undifferentiated carcinoma (1 case) and adenoid cystic carcinoma (2 cases) did not possess detectable ABO (H) antigens in neoplastic areas of the gland. The absence of ABO antigens in normal acinar glands supports their suggested myoepithelial or mesenchymal derivation, as the absence of antigen in benign and malignant mixed tumors supports their proposed mesenchymal derivation. Ductular epithelium and the epithelial components of benign Warthin's tumors have ABO (H) antigens, while the loss of antigen in the epithelial portion of the malignant Warthin's tumor is characteristic of epithelial neoplastic dedifferentiation. Loss of antigen in adenoid cystic and undifferentiated carcinomas of the parotid supports the concept that antigen is absent in epithelially derived malignant neoplasms.

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.

Direct immunofluorescence for ABH blood group isoantigens: Use of FITC-conjugated lectins

A direct immunofluorescence method was used to detect A, B, or H blood group isoantigens in deparaffinized tissue sections of 30 bladder tumors of various grades and stages. The results were compared with those of the specific red cell adherence test performed on the replicate sections. The two methods showed identical results for isoantigens A, B, and H. Direct immunofluoresence method appeared to be less time-consuming, more simple to perform, and reproducible with less subjectivity in the interpretation of the results.

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.

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.

The Specific Red Cell Adherence Test in Transitional Cell Carcinoma of the Bladder Before and After Radiotherapy in Patients With Blood Group A

Serial formalin-fixed specimens of bladder tumors before and after irradiation from 19 patients with blood group A were tested for the presence or absence of antigen by the the specific red cell adherence test. All patients had at least grade B transitional cell cancer of the bladder. The specific red cell adherence test was negative before radiotherapy in 9 patients, 4 of whom (44 per cent) had positive tests after treatment. However, the test was positive before radiotherapy in 10 patients (56 per cent) and became negative after radiotherapy in only 1. Of the patients 47 per cent had tumors that were positive for specific red cell adherence before and after radiotherapy, and 26 per cent had tumors that were consistently negative for antigen.

Urothelium and the Specific Red Cell Adherence Test.

The Specific Red Cell Adherence Test (SRCAT) has been modified to produce clearer, more consistent, permanent preparations. Normal urothelium, inflammatory lesions, squamous metaplasia, papillomata, flat in situ and papillary tumours and invasive neoplasms of the bladder have been studied. Inflammatory and squamous changes did not corrupt the test results. The SRCAT was found to be a highly significant test for identifying those patients at risk from invasive bladder tumour. ABH antigens in carcinoma in situ appeared to be non-uniformly distributed throughout the bladder urothelium; it is suggested that all subsequent biopsies from these patients should be tested.