Direct cytocidal effect of galectin-9 localized on collagen matrices on human immune cell lines

• Published in: Biochimica et biophysica acta Vol. 1840; no. 6; pp. 1892 - 1901
• Main Authors: Fukata, Youko, Itoh, Aiko, Nonaka, Yasuhiro, Ogawa, Takashi, Nakamura, Takanori, Matsushita, Osamu, Nishi, Nozomu
• Format: Journal Article
• Language: English
• Published: Netherlands 01.06.2014
• Subjects: allografting; autoimmune diseases; Binding Sites; cell death; Cell Proliferation - drug effects; collagen; Collagen - metabolism; collagenase; culture media; disease models; Galectins - chemistry; Galectins - pharmacology; human cell lines; Humans; immunosuppressive agents; Immunosuppressive Agents - pharmacology; Jurkat Cells; lectins; organ transplantation; Protein Structure, Tertiary; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - pharmacology; T-lymphocytes; Galectin; Immune suppression; Collagen-binding domain; Collagen
• ISSN: 0304-4165; 0006-3002
• DOI: 10.1016/j.bbagen.2014.01.019

There is a continuous demand for new immunosuppressive agents for organ transplantation. Galectin-9, a member of the galactoside-binding animal lectin family, has been shown to suppress pathogenic T-cell responses in autoimmune disease models and experimental allograft transplantation. In this study, an attempt has been made to develop new collagen matrices, which can cause local, contact-dependent immune suppression, using galectin-9 and collagen-binding galectin-9 fusion proteins as active ingredients. Galectin-9 and galectin-9 fusion proteins having collagen-binding domains (CBDs) derived from bacterial collagenases and a collagen-binding peptide (CBP) were tested for their ability to bind to collagen matrices, and to induce Jurkat cell death in solution and in the collagen-bound state. Galectin-9-CBD fusion proteins exhibited collagen-binding activity comparable to or lower than that of the respective CBDs, while their cytocidal activity toward Jurkat cells in solution was 80~10% that of galectin-9. Galectin-9 itself exhibited oligosaccharide-dependent collagen-binding activity. The growth of Jurkat cells cultured on collagen membranes treated with galectin-9 was inhibited by~90%. The effect was dependent on direct cell-to-membrane contact. Galectin-9-CBD/CBP fusion proteins bound to collagen membranes via CBD/CBP moieties showed a low or negligible effect on Jurkat cell growth. Among the proteins tested, galectin-9 exhibited the highest cytocidal effect on Jurkat cells in the collagen-bound state. The effect was not due to galectin-9 released into the culture medium but was dependent on direct cell-to-membrane contact. The study demonstrates the possible use of galectin-9-modified collagen matrices for local, contact-dependent immune suppression in transplantation.