SUMMARYThe precursors of the blood group N and M-immunodominant structures, Tn and T (Thomsen-Friedenreich) epitopes (EPs) occur in ف 90% of carcinomas (CAs) but are masked in benign-diseased and healthy tissues. We determined quantitatively on 55 primary invasive ductal breast CAs, stages I to IV, the prognostic value of extent of Tn and T EP expression over an observation period exceeding 5 years postoperatively. Classical, established pathological and histological prognostic characteristic indicators associated with survival were subdivided by standard criteria into favorable and unfavorable categories. Tissue sections were reacted with monoclonal anti-Tn and -T antibodies, followed by the streptavidin-biotin-peroxidase-DAB procedure; counterstain was methyl green. Tn and T EPs were then quantitated by computerized image analysis. Of the 55 CAs, 51 clearly expressed Tn and T, and four had traces. Strong Tn EP expression was statistically significantly associated with shortened 5-year disease-free interval, increasing pTNM stages, positive lymph node status, and increasing combined histological grades. T EPs were usually well expressed but showed no significant association with prognostic factors. Our results suggest that quantitative immunohistochemistry-image analysis of Tn EPs of primary breast CAs may add new parameters to prognostication. (J Histochem Cytochem 45:1393-1400, 1997)
The composition of tumor-infiltrating lymphocytes (TIL) often reflects the host's immune response to the tumor. To study the relationship of TIL and carcinoma-associated T/Tn antigens in breast carcinoma, a straightforward concurrent immunoenzyme staining procedure was developed. Fresh tissue was directly fixed in a zinc-based fixative to preserve lymphocyte markers and then routinely embedded in paraffin. The TIL subtypes in the sections were identified in the first immunostaining cycle by reaction with a monoclonal antibody (MAb) to lymphocyte markers CD3, CD4, CD8, CD19, or CD56, followed by a modified avidin-biotin procedure and diaminobenzidine tetrahydrochloride-H2O2 for color development. This was followed by paraformaldehyde fixation to block antibody crossreactivity. The T and Tn antigens on carcinoma cells were then demonstrated in a second staining cycle by reaction with an MAb against T or Tn antigen, followed by an indirect immunoalkaline phosphatase procedure and corresponding substrate systems for color development. The distinguishable brown color for TIL and blue or red color for T or Tn antigen enabled us to identify the TIL subsets and to describe their relations with T/Tn antigen expression in situ. This approach may contribute to better understanding of the patients' immune defenses against their tumor and aid in prognostication.
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