期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 367, 期 1-2, 页码 56-62出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2011.02.001
关键词
Autoantibodies; Cytokine; Vaccination
资金
- FRIA (Fonds pour la Formation et la Recherche dans l'Industrie et dans l'Agriculture, Brussels, Belgium)
- Belgian State, Prime Minister's Office
- Science Policy Programming
- Actions de Recherche Concertees of the Communaute Francaise de Belgique
Induction of autoantibodies towards immune regulatory proteins, such as cytokines or their receptors, is a powerful strategy for functional studies on the role of these factors in vivo. Here we describe a new procedure to elicit autoantibodies by taking advantage of tumor cells as a vaccine against peptides presented at their surface in fusion with the human CD134L transmembrane protein. P1.FITR, an immunogenic variant of the P815 mastocytoma cell line, was used to generate stably transfected cell clones with expression vectors encoding the human CD134L transmembrane protein fused with either mouse IL-22BP or IL-9. Following repeated injections of living tumor cells expressing the mIL-228P construct, mice developed autoantibodies that bind to mIL-228P and inhibit its interaction with IL-22 in vitro. Mice similarly immunized against mIL-9 produced high titers of autoantibodies that block the activity of this cytokine in the TS1 bioassay. This procedure also inhibits IL-9 activity in vivo as no increase of serum MMCP-1 mast cell protease concentration was observed following IL-9 administration to immunized mice. As an alternative to the injection of living tumor cells expressing the CD134L-antigen fusion protein, intramuscular electrotransfer of the corresponding DNA construct also induced autoantibodies. These results validate this method as a simple and convenient approach to knock down the in vivo activity of soluble regulatory proteins, including cytokines and their receptors. (C) 2011 Elsevier B.V. All rights reserved.
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