期刊
JOURNAL OF LEUKOCYTE BIOLOGY
卷 89, 期 6, 页码 1001-1007出版社
WILEY
DOI: 10.1189/jlb.1210699
关键词
vaccine; interleukin; chemokine; cross-linking; antigen
资金
- Fonds National de la Recherche Scientifique Medicale (FRSM, Belgique)
- Concerted Research Actions (G.O.A.) of the Regional Government of Flanders
- Belgian Federal Science Policy
- Swiss National Foundation [310030-129852/1]
- Swiss National Science Foundation (SNF) [310030_129852] Funding Source: Swiss National Science Foundation (SNF)
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-beta 1, 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. J. Leukoc. Biol. 89: 1001-1007; 2011.
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