alexa Anti-Thomsen-Friedenreich (T) antibody-based ELISA and its application to human breast carcinoma detection.


Journal of Clinical & Cellular Immunology

Author(s): Desai PR, Ujjainwala LH, Carlstedt SC, Springer GF

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Abstract An anti-Thomsen-Friedenreich (T) antibody-based enzyme-linked immunosorbent assay (ELISA) with high efficacy in human breast carcinoma detection is described. Immunoreactive T epitopes occur in approximately 90\% of all carcinomata; all humans have anti-T antibodies, naturally occurring anti-carcinoma antibodies, induced by their own intestinal flora. Carcinoma patients, but not control subjects, show alterations of serum anti-T hemagglutinin levels. Human anti-T antibodies are predominantly IgM. In the protocol presented here, anti-T IgM antibodies are quantitated 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 control subjects, use of the quotient, QMe, which also considers total IgM, increases this distinction. For a given serum, QMe was obtained by the formula: QMe = (100 x (anti-T IgM)2/total IgM). Sera of 242 subjects, 117 breast carcinoma patients, 36 benign breast disease patients and 89 healthy persons were analyzed. QMe identified 88\% of the breast carcinoma patients: it all six (100\%) in situ, 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 patients 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 subjects were negative by both anti-T IgM and QMe. In preliminary studies, the ELISA protocol detected 11/14 (79\%) patients with carcinomata other than those of the breast. The identification of all six in situ breast carcinoma patients by QMe points to its usefulness in carcinoma detection, especially early.
This article was published in J Immunol Methods and referenced in Journal of Clinical & Cellular Immunology

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