Amine-reactive OVA multimers for auto-vaccination against cytokines and other mediators: perspectives illustrated for GCP-2 in L. major infection

• Published in: Journal of Leukocyte Biology Vol. 89; no. 6; pp. 1001 - 7
• Main Authors: Uyttenhove, Catherine, Marillier, Reece G, Tacchini-Cottier, Fabienne, Charmoy, Mélanie, Caspi, Rachel R, Damsker, Jesse M, Goriely, Stanislas, Su, Dan, Van Damme, Jozef, Struyf, Sofie, Opdenakker, Ghislain, Van Snick, Jacques
• Format: Journal Article
• Language: English
• Published: Liss 01.06.2011
• ISSN: 0741-5400

Anticytokine auto-vaccination is a powerful tool for the study of cytokine functions in vivo but has remained rather esoteric as a result of numerous technical difficulties. We here describe a two-step procedure based on the use of OVA multimers purified by size exclusion chromatography after incubation with glutaraldehyde at pH 6. When such polymers are incubated with a target protein at pH 8.5 to deprotonate reactive amines, complexes are formed that confer immunogenicity to self-antigens. The chemokine GCP-2/CXCL6, the cytokines GM-CSF, IL-17F, IL-17E/IL-25, IL-27, and TGF-β1, and the MMP-9/gelatinase B are discussed as examples. mAb, derived from such immunized mice, have obvious advantages for in vivo studies of the target proteins. Using a mAb against GCP-2, obtained by the method described here, we provide the first demonstration of the major role played by this chemokine in rapid neutrophil mobilization after Leishmania major infection. Pre-activated OVA multimers reactive with amine residues thus provide an efficient carrier for auto-vaccination against 9-90 kDa autologous proteins.