2014 Fiscal Year Research-status Report
魚類免疫機構における重要なサイトカインIL-2/15/15Lファミリーの解明
| Project/Area Number |
25450310
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| Research Institution | Fujita Health University |
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Principal Investigator |
J・M Dijkstra 藤田保健衛生大学, 総合医科学研究所, 講師 (10387681)
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | IL-15-like / IL-15 / IL-2 / Fish / Immunology / Evolution |
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| Outline of Annual Research Achievements |
The major experimental achievement this year is that we found that the rainbow trout cytokines IL-2, IL-15, and IL-15L have functional effects on trout lymphocytes. We are currently writing those results into an article. The function of IL-15L we like to keep secret here, because it concerns a newly discovered cytokine. Important for vaccine adjuvants purposes is that we found that trout IL-2 and IL-15 can stimulate cytotoxic cells if presented together with IL-15Ra, which agrees with expectations based on studies in mammals. Although for bovine IL-2, IL-15 and IL-21 we could find the expected functions, we have not been able so far to find a function for bovine IL-15L.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
We are very glad that all the trout and bovine cytokines that we expressed recombinantly, with the exception of bovine IL-15L, are functional and able to affect lymphocytes. The writing of those data into a publication is in good progress. However, the expression of soluble proteins in other systems than mammalian cells takes more time than we expected.
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| Strategy for Future Research Activity |
The accumulating trout data are most promising and important to us, so we will continue analyzing the trout IL-2, IL-15 and IL-15L molecules. So far we have worked with the recombinant trout proteins from the supernatants of transfected mammalian cells, which is good in the sense that these proteins should be properly folded. But we need higher amounts for efficient analysis of various functions, including vaccine adjuvant properties. Therefore this year will be especially important for the recombinant production of the trout cytokines in other expression systems such as E.coli and insect cells. This will include free cytokines, as well as cytokines genetically fused to IL-15Ra. The assays in which the recombinant cytokines will be tested will be biased towards finding the best conditions for stimulation of cytotoxic T cells. We hope to achieve similar results as those that we obtained with the trout cytokines expressed by transfected mammalian cells.
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