| 研究概要 |
This year included a lot of paper writing, which resulted in three publications.
Publication 1, in which we described the evolution of cytokine IL-15L and its binding to receptor chain IL-15Ra: Dijkstra JM, et al., 2014, Identification of a gene for an ancient cytokine, interleukin 15-like, in mammals; interleukins 2 and 15 co-evolved with this third family member, all sharing binding motifs for IL-15Rα. Immunogenetics 66:93-103. Publication 2, which comprehensively described the MHC class II situation in fish, which is necessary for intelligent manipulation of T helper cells: Dijkstra JM,et al., 2013, Comprehensive analysis of MHC class II genes in teleost fish genomes reveals dispensability of the peptide-loading DM system in a large part of vertebrates. BMC Evol Biol 13:260. Publication 3 includes discussion of the evolution of the interleukins 2, 15 and 15L : Dijkstra JM. Title may be changed by the editors. Nature, brief communications arising, in press.
The major experimental achievement this year is that we found that the cytokines IL-2, IL-15, and IL-15L cross-react with IL-15Ra across fish-mammal borders, except for mammalian IL-2 which only binds to mammalian IL-2Ra. Furthermore, we made expression constructs for the rainbow trout and bovine receptor chains IL-2Rb, IL-2Rg and IL-21Ra, and will soon analyze their stimulation by the several cytokines.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
With three papers as first author a scientist should never complain. The binding studies of cytokines IL-2, IL-15 and
IL-15L to receptor chain IL-15Ra, as measured by FACS analysis, also went very fine. For rainbow trout now both
the cytokine part and the receptor part are fully prepared for functional analysis of the cytokines. Unfortunately the carp and ginbuna work did not show important progress, and neither did the bacterial expression studies as we still face solubility problems. We did however, manage to copy a soluble human IL-15Ra-IL-15 fusion published by others, and now hope to make a similar fusion for IL-15L. Also because writing the papers took a lot of time, we are quite satisfied that at least for the trout system we made important progress.
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| 今後の研究の推進方策 |
Plan for fiscal year 2014 (01-04-2014 to 01-04-2015)
The plan for trout is to study the function of the cytokines IL-2, IL-15 and IL-15L, alone and in combination with soluble IL-15Ra. With which receptor chains (other than IL-15Ra) do they interact, and which cells do they stimulate? Depending on the results of those experiments, for which we initially will use a recombinant eukaryotic expression system, we will choose the most promising cytokine (+ soluble IL-15Ra) for cheap production in bacteria, yeast and/or insect cells. Those will be used as vaccine adjuvants. We will then also check cross-reactivity of the recombinant trout cytokines with the immune system of other fish species. Meanwhile we will try to improve the current data on carp and ginbuna IL-2, IL-15 and IL-15L. Whether we can perform surface plasmon resonance (SPR) for cytokine to IL-15Ra binding analysis depends on if we can tackle the solubility problem of IL-15L (which seems highly unstable without IL-15Ra).
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| 次年度の研究費の使用計画 |
Some money was shifted across the fiscal year border. The main reason for that is that the color charges for one of our publications, that we intended to pay with the 2013 budget, were charged from our account only months after the actual publication.
The extra money of the 2013 budget left after paying the above mentioned color charges will be used in 2014 for ongoing laboratory costs (kits etc.)
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