Kadoshima-Yamaoka Kumiko, Murakawa Masao, Goto Megumi, Tanaka Yoshitaka, Inoue Hidekazu, Murafuji Hidenobu, Hayashi Yasuhiro, Nagahira Kazuhiro, Miura Kenju, Nakatsuka Takashi, Chamoto Kenji, Fukuda Yoshiaki, Nishimura Takashi
Biomedical Research Laboratories, Asubio Pharma Co., Limited, Osaka 618-8503, Japan.
Int Immunopharmacol. 2009 Jan;9(1):97-102. doi: 10.1016/j.intimp.2008.10.005. Epub 2008 Nov 6.
In the present study, possible role of phosphodiesterase 7 (PDE7) in development and function of cytotoxic T lymphocyte (CTL) was examined using ASB16165, a specific inhibitor for PDE7. ASB16165 inhibited generation of CTL activity in mixed lymphocyte reaction (MLR), in which splenocytes from C57BL/6N mice were stimulated with those from BALB/c mice. Flow cytometric analysis revealed that ASB16165 suppressed induction of activated CD4+ as well as CD8+ T cells in MLR. In cell division analyses using 5-carboxyfluorescein diacetate succinimide ester (CFSE), ASB16165 was shown to markedly inhibit proliferation of CD4+ and CD8+ T cells. In addition, ASB16165 reduced effector function of CTL, while the effect was less than that observed in CTL induction in MLR. Forskolin and dibutyryl cAMP also inhibited both the induction and effector function of CTL. PDE4 inhibitor rolipram showed similar but weaker inhibition for the development and proliferation of CD8+ T cells compared with ASB16165, and failed to impair effector function of CTL. These findings suggest that PDE7 but not PDE4 has the major role in induction and function of CTL in mice, and that the effect might be mediated by elevation of intracellular cAMP level. ASB16165 may be useful for treatment of the diseases in which CTL has a pathogenic role (e.g. autoimmune diseases).
在本研究中，使用磷酸二酯酶7（PDE7）的特异性抑制剂ASB16165研究了PDE7在细胞毒性T淋巴细胞（CTL）发育和功能中的可能作用。ASB16165抑制了混合淋巴细胞反应（MLR）中CTL活性的产生，在该反应中，用BALB/c小鼠的脾细胞刺激C57BL/6N小鼠的脾细胞。流式细胞术分析显示，ASB16165抑制了MLR中活化的CD4+以及CD8+T细胞的诱导。在使用5-羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）的细胞分裂分析中，ASB16165被证明可显著抑制CD4+和CD8+T细胞的增殖。此外，ASB16165降低了CTL的效应功能，但其作用小于在MLR中CTL诱导时观察到的作用。福斯可林和二丁酰环磷腺苷也抑制了CTL的诱导和效应功能。与ASB16165相比，PDE4抑制剂咯利普兰对CD8+T细胞的发育和增殖表现出相似但较弱的抑制作用，并且未能损害CTL的效应功能。这些发现表明，PDE7而非PDE4在小鼠CTL的诱导和功能中起主要作用，并且该作用可能由细胞内cAMP水平升高介导。ASB16165可能对治疗CTL起致病作用的疾病（例如自身免疫性疾病）有用。
