The chemokine CXCL10 is released by a plethora of cells, including immune and metastatic cancer cells, following stimulation with interferon-gamma. It acts via its GPC receptor on T-cells attracting them to various target tissues. Glycosaminoglycans (GAGs) are regarded as co-receptors of chemokines, which enable the establishment of a chemotactic gradient for target cell migration. We have engineered human CXCL10 towards improved T-cell mobilisation by implementing a single site-directed mutation N20K into the protein, which leads to a higher GAG binding affinity compared to the wild type. Interestingly, this mutation not only increased T-cell migration in a transendothelial migration assay, the mutant intensified T-cell chemotaxis also in a Boyden chamber set-up thereby indicating a strong role of T-cell-localised GAGs on leukocyte migration. A CXCL10 mutant with increased GAG-binding affinity could therefore potentially serve as a T-cell mobiliser in pathological conditions where the immune surveillance of the target tissue is impaired, as is the case for most solid tumors.
Protein eng des sel
367 - 373
CXCL10, T-cell immobiliser, chemokines, glycosaminoglycan, surface plasmon resonance, Amino Acid Sequence, Binding, Competitive, Cell Movement, Cells, Cultured, Chemokine CXCL10, Chemotaxis, Leukocyte, Glycosaminoglycans, Humans, Male, Molecular Dynamics Simulation, Mutation, Missense, Protein Binding, Protein Conformation, Protein Engineering, T-Lymphocytes