We studied the blood specimens from 117 women with malignant and benign breast diseases that obtained diagnostics and treatment in the breast disease department of the Ukrainian Institute of the Oncology and Radiology, Kiev. Besides this the blood of 21 healthy donors was studied. The age of patient and donors was 27 – 87 years. Cancer diagnosis was approved by hystological method, diagnose of benign tumor and mastopathy was approved by hystological, cytological and clinical methods. In patient group there were 872 women with primary diagnosis of breast cancer of stage I – IY, including 14 patients after presurgery radiation.; 21 patient with benign breast diseases (fibroadenoma, fibroadenomatosis), 26 patients after complex treatment (surgery, radio- and chemotherapy) and 16 patients with non-breast cancer.

EM-test was performed according to Broen et. al. /3/ with own modifications /3/. Nearaminidase-treated antigenic membrane preparation were obtained from normal and malignant human breast tissues. Test results were supposed to be positive if the decrease of the electrophoretic mobility of the indicator cells (EM-indices) was equal and/or more than 10% from the control.

The initial step of the clinical investigations was the determination of the operational characteristics of EM-indices, distinguishing positive and negative results. In the study with breast cancer patients and healthy donors (men) it was shown that the test specificity was 83% and sensitivity – 90%: positive prognostic value of 98% and negative prognostic value of 65,5%.

Sensitization to normal desialilated antigens (NDSAg) and cancer desialilated antigens (CaDSAg) were compared in the patients with different tumor stage. The results are shown in the table.

Ag               Breast cancer stages 
                    I          II          III         IY       All                                      
NDSAg
Positive/total, %   2/2      20/29        18/18      10/11     48/58  
Sensitivity        100%       69%         100%        91%       83%
EM index, %         85     29.9±6.0      56.3±7.5    37.7±6.3  
CaDSAg
Positive/total, %   -        13/18        9/11        4/4      26/33  
Sensitivity         -         72%          82%        100%      78%
EM index, %         -     18,9±3,8      31,2±8,2     52,3±14,3   
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Next study was to compare the results of EM-testing in patient with- and without pre-surgery irradiation. We detected the correlation between cytocydic effect of radiation in some patients and decline in EM-indices. EM-indices also correlated with the mass volume (for T₂ and T₃ (P <: ,05). No correlation has been found between EM-indices and the hystolohical grade of malignancy, lymph nodes involvement, methastatic process.

During the study of the lymphocyte sensitization in patients with benign breast diseases it was demonstrated the positive EM-test reaction in 7/9 patients with fibroadenoma (77.8%) and in 8/13 (61,55) patients with fibroadenomatosis (in North America – non-dominant lesion) with reliable difference between these two groups (p <: .05). We suppose this difference is much more like due to the mass volume, than to the different cancer potential of these forms.

The study of the prognostic capacity of NDSAg in recurrence prediction in breast cancer patients after the combined treatment has been held in 26 patients. 50% showed the positive reaction in the range of 13 - 82%. We succeeded to follow up 16 patients during four years: in 13 of them the EM-test indices correlated with the current of the disease. In several cases the positive reaction preceded the clinical manifestation of the metastasis for 1 - 3 years.

The results, shown above, summarize total of 18 years of study of the normal desialilated antigens. Initial purpose of the work was the augmentation of tumor cells’ antigenicity by neuraminidase treatment. As it revealed, neuraminidase treatment in special conditions augments not only antigens in the tumor tissue, but also unmasks hidden antigenic determinants on the normal cell surface, that are intimately involved in cancerogenesis in its early stages. Desialilated membrane antigens can be used in the heterogenous systems, that provides the possibility for their application for human cancer immunodetection.

The application of normal desialilated antigens is the most promising for preclinical screening of real, evidence-based high risk groups, were the regular follow-up and possible prescription of such prophylaxis drugs like Tamoxifen and Raloxifen can bring the immediate results by increasing health care standards. Rather time and labor-consuming EM-test procedure may be avoided by the replacement of electrophoretic mobility measurements by lymphokine determination in ELISA test.

1. Springer G.F.(1983) Triple role of T and Tn antigens in cancer: as universal clonal carcinoma markers, in cancer cell adhesion and as autoimmunogens. // Surv. Synth. Path. Res. - 2. - P.141-164.
2. Novichenko N. (1985) Histospecific tumor-associated antigens from normal and malignant tissues treated with viral neuraminidase: their application in the electrophoretic mobility test (EM-test) . //Cell Electrophoresis, Walter de Grueter, - Berlin – New-York. – P.861-869.
3. Broen B.von, Pasternak G., Le than Thuy et. al. (1976) Untersuchungen uber des zell-elektrophorese-mobilitats-methode // DDR Med. Rep. - 5. - N10. - P.910-914.