ANTI-THOMSEN-FRIEDENREICH (T)-ANTIBODY-BASED ELISA AND ITS APPLICATION TO HUMAN BREAST-CARCINOMA DETECTION

An anti-Thomsen-Friedenreich (T) antibody-based enzyme-linked immunoso rbent assay (ELISA) with high efficacy in human breast carcinoma detec tion is described. Immunoreactive T epitopes occur in similar to 90% o f all carcinomata; all humans have anti-T antibodies, naturally occurr ing anti-carcinoma antibodies, induced by their own intestinal flora. Carcinoma patients, but not control subjects, show alterations of seru m anti-T hemagglutinin levels, Human anti-T antibodies are predominant ly IgM. In the protocol presented here, anti-T IgM antibodies are quan titated by ELISA using Immulon 2 wells coated with human blood group O erythrocyte-derived T antigen as solid phase; in addition, total IgM in each serum is quantitated by ELISA in parallel with the anti-T IgM. Inter-assay coefficient of variation was 2% for both ELISAs. Although anti-T IgM values alone distinguish between carcinoma patients and co ntrol subjects, use of the quotient, Q(Me), which also considers total IgM, increases this distinction. For a given serum, Q(Me) was obtaine d by the formula: Q(Me) = (100 X (anti-T IgM)(2)/total IgM). Sera of 2 42 subjects, 117 breast carcinoma patients, 36 benign breast disease p atients and 89 healthy persons were analyzed. Q(Me) identified 88% of the breast carcinoma patients: it was positive in all six (100%) in si tu, 11/13 (85%) Stage I, 48/58 (83%) Stage II and III and 38/40 (95%) Stage IV patients, Sera from 83% of the 36 benign breast disease patie nts were negative, i.e. within normal range; five of the six positive sera originated from patients with increased long-term risk of breast carcinoma, while sera from 11 other patients with increased carcinoma risk were negative. Overall, 90% of the 125 non-carcinoma control subj ects were negative by both anti-T IgM and Q(Me). In preliminary studie s, the ELISA protocol detected 11/14 (79%) patients with carcinomata o ther than those of the breast. The identification of all six in situ b reast carcinoma patients by Q(Me) points to its usefulness in carcinom a detection, especially early.