A newly generated functional antibody identifies Tn antigen as a novel determinant in the cancer cell-lymphatic endothelium interaction

Carla Danussi, Anna Coslovi, Cristiana Campa, Maria T. Mucignat, Paola Spessotto, Fulvio Uggeri, Sergio Paoletti, Alfonso Colombatti

Research output: Contribution to journalArticle

Abstract

Malignant transformation of epithelial cells is frequently associated with the alteration of glycosylation pathways. Tn is a common tumor-associated carbohydrate antigen present in 90% of human carcinomas and its expression correlates with metastatic potential and poor prognosis. Despite its relevance, the functional role of Tn in tumor biology has not been firmly established probably for the lack of appropriate experimental tools. Our aims were to produce highly reactive monoclonal antibodies against Tn making use of synthetically produced Tn and to test their usefulness for in vivo imaging as well as to define their potential functional activity in tumor cell spread. We immunized mice with Tn clustered on cationized BSA and screened the positive hybridomas with Tn-biotinylated alginate. Enzyme-linked immuno sorbent assay and immunofluorescence assays revealed that the most reactive anti-Tn IgM mAb (2154F12A4) selectively recognized Tn on the MCF7 breast cancer cell line since its binding to the cell membrane was completely abolished by preincubation with purified Tn. Importantly, QDot 800-conjugated mAb injected in MCF7-tumor bearing mice specifically bound to primary tumor lesions as well as to metastases in lymph nodes. In addition, this mAb was able to inhibit cancer cell adhesion to lymphatic endothelium suggesting a novel involvement of Tn in the lymphatic dissemination of cancer cells and hypothesizing future applications in inhibiting lymphatic metastases.

Original languageEnglish
Pages (from-to)1056-1067
Number of pages12
JournalGlycobiology
Volume19
Issue number10
DOIs
Publication statusPublished - 2009

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Lymphatic Endothelium
Tumors
Cells
Antibodies
Assays
Neoplasms
Bearings (structural)
Tumor-Associated Carbohydrate Antigens
Glycosylation
Cell adhesion
Cell membranes
Sorbents
Lymphatic Metastasis
Immunoglobulin M
Monoclonal Antibodies
Hybridomas
Imaging techniques
Cell Adhesion
Fluorescent Antibody Technique
Tn antigen

Keywords

  • In vivo imaging
  • Lymphatic endothelial cells
  • Metastasis
  • Monoclonal antibody
  • Tn antigen

ASJC Scopus subject areas

  • Biochemistry

Cite this

A newly generated functional antibody identifies Tn antigen as a novel determinant in the cancer cell-lymphatic endothelium interaction. / Danussi, Carla; Coslovi, Anna; Campa, Cristiana; Mucignat, Maria T.; Spessotto, Paola; Uggeri, Fulvio; Paoletti, Sergio; Colombatti, Alfonso.

In: Glycobiology, Vol. 19, No. 10, 2009, p. 1056-1067.

Research output: Contribution to journalArticle

Danussi, Carla ; Coslovi, Anna ; Campa, Cristiana ; Mucignat, Maria T. ; Spessotto, Paola ; Uggeri, Fulvio ; Paoletti, Sergio ; Colombatti, Alfonso. / A newly generated functional antibody identifies Tn antigen as a novel determinant in the cancer cell-lymphatic endothelium interaction. In: Glycobiology. 2009 ; Vol. 19, No. 10. pp. 1056-1067.
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