Correlative Fine Specificity of Several Thomsen-Friedenreich Disaccharide-Binding Proteins with an Effect on Tumor Cell Proliferation

Abstract

Epithelial cancer cells show increased cell surface expression of mucin antigens with aberrant O-glycosylation, notably type I core (Galbeta1-3GalNAcalpha), termed Thomsen-Friedenreich disaccharide (TFD), a chemically well-defined carbohydrate antigen with a proven link to malignancy. Several TFD-binding proteins influence the proliferation of cells to which they bind. We studied the fine specificity of TFD-binding proteins and its relationship with epithelial tumor cell proliferation. Competitive binding assays against asialoglycophorin showed that Agaricus bisporus lectin (ABL) and human anti-TFD monoclonal antibody (mAb) TF1 were inhibited only by TFD and its alpha-derivatives. Peanut agglutinin (PNA), mAb TF2, and mAb TF5 were also inhibited by other carbohydrates such as lacto-N-biose (Galbeta1-3GlcNAc), lactose, and (Mealpha or beta) Gal, indicating lower recognition of the axial C-4 hydroxyl group position of GalNAc from TFD, and the major relevance of the terminal Gal on interaction of these three TFD-binding proteins. In the direct glycolipid-binding assay, ABL bound mostly to alpha-anomeric TFD-bearing glycolipids, whereas PNA interacted mainly with beta-linked TFD. Of the three anti-TFD mAbs analyzed, all bound N5b (terminal beta-TFD), but only TF2 interacted with N6 (terminal alpha-TFD). These findings indicate that TFD-binding proteins that stimulate the proliferation of epithelial tumor cell lines recognize mainly a terminal beta-Gal region of beta-linked TFD, whereas ABL, which inhibits the proliferation of these tumor cells, binds mainly to subterminal GalNAc of alpha-anomeric TFD.

Identification of Galectin-3 and Mucin-Type O-Glycans in Breast Cancer and Its Metastasis to Brain

“…ALL is specific for a special GalNAc-containing motif found in O-glycoproteins (26). The T and Tn antigens have been suggested to be used in early detection screening for breast cancer (27)(28)(29) and for hepatocellular cancer (30,31), and they have been considered to be associated with tumors with high risk to metastasize (32). In healthy individuals, these antigens have been also associated with immature cells; moreover, it has been identified that ALL + cell populations possess higher resistance to apoptosis.…”

Preclinical Studies with JAA-F11 Anti-Thomsen-Friedenreich Monoclonal Antibody for Human Breast Cancer

“…In support of the EIA data for detecting TF-Ag-positive cells, multiple other studies have shown that approximately 80% of human breast carcinomas are positive for TF-Ag [17,[31][32][33][34][35][36], usually by immunohistochemistry. Not all anti-TF-Ag antibodies are useful in immunohistochemistry, and JAA-F11 currently fits in this group, although efforts are being made in our laboratory to develop appropriate fixation methods for immunohistochemistry with JAA-F11.…”

“…The ER + , PR + and HER2 -cell line T47D also showed a decrease in proliferation at all concentrations of JAA-F11, which reached significance at 2 and 4 μg/ml. PNA (Arachishypogaea) is a lectin that has long been used for the detection of TF-Ag on the surface of carcinoma cells [35,36]. PNA has been shown to increase proliferation of TF-Ag-bearing tumor cells [36].…”

“…PNA (Arachishypogaea) is a lectin that has long been used for the detection of TF-Ag on the surface of carcinoma cells [35,36]. PNA has been shown to increase proliferation of TF-Ag-bearing tumor cells [36]. Owing to this proliferation-stimulating characteristic, it was chosen as a treatment to compare the effect of JAA-F11 on tumor cells.…”

Binding of galectin-1 (gal-1) to the Thomsen–Friedenreich (TF) antigen on trophoblast cells and inhibition of proliferation of trophoblast tumor cells in vitro by gal-1 or an anti-TF antibody

“…Several TF-binding proteins influence epithelial tumor cell proliferation (Yu et al 1997). TF-disaccharide binding proteins, that stimulate the proliferation of epithelial tumor cell lines, recognize mainly the terminal b-Gal region of b-linked TF antigen, whereas proliferation inhibition is mainly associated by binding to subterminal GalNAc of a-anomeric TF antigen (Irazoqui et al 2001). On tumor cells MUC1 is post translationally modified resulting in incomplete O-glycosylation and presentation of the TF-disaccharide.…”