Amine-reactive OVA multimers for auto-vaccination against cytokines and other mediators: perspectives illustrated for GCP-2 inL. majorinfection

Abstract

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.