| Summary: | 碩士 === 國立陽明大學 === 微生物及免疫學研究所 === 99 === TREM-DAP12 protein complex on myeloid-derived innate immune cell surface participates in immune modulation, bone homeostasis, and neural development. Recently, the identification of PlexinA1-TREM2-DAP12 signaling complex in dendritic cells provides an expanding role of TREM family as adaptor proteins for other surface receptors. We speculate that some innate receptors might mediate activating signals through associating with TREM-DAP12 complex in macrophages. Here, we established a screening system for TREM dependent, DAP12–associated receptor complex to identify novel receptors functioning through TREM-DAP12 signaling in mouse macrophages. Expression vectors for DAP12, TREM1, TREM2 and TREM3 were generated. Furthermore, TREM-DAP12 receptor complex pull down system was established and applied in DAP12-overexpressed wild type and various TREM deficient macrophages. The DAP12 pull-down pattern changes found in TREM deficient cells will provide insight to identify novel TREM-associated receptors that through DAP12 for signaling. Taken together, we identified DAP12 overexpression could lead to macrophage immortalization. We generate WT and TREM knockout transformed macrophages cell lines for biochemistry studies. The TREM-DAP12 associated receptors were purified and identified with mass spectrometry and will then goes for further functional analysis.
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