Graduate School of Medicine, Kobe University, Kobe, 650-0017, Japan.
Center for Innovation in Immunoregulative Technology and Therapeutics, Kyoto University Graduate School of Medicine, Kyoto, Japan.
Sci Rep. 2017 Aug 9;7(1):7663. doi: 10.1038/s41598-017-07936-9.
Knock-in mice lacking PKN1 kinase activity were generated by introducing a T778A point mutation in the catalytic domain. PKN1[T778A] mutant mice developed to adulthood without apparent external abnormalities, but exhibited lower T and B lymphocyte counts in the peripheral blood than those of wild-type (WT) mice. T and B cell development proceeded in an apparently normal fashion in bone marrow and thymus of PKN1[T778A] mice, however, the number of T and B cell counts were significantly higher in the lymph nodes and spleen of mutant mice in those of WT mice. After transfusion into WT recipients, EGFP-labelled PKN1[T778A] donor lymphocytes were significantly less abundant in the peripheral circulation and more abundant in the spleen and lymph nodes of recipient mice compared with EGFP-labelled WT donor lymphocytes, likely reflecting lymphocyte sequestration in the spleen and lymph nodes in a cell-autonomous fashion. PKN1[T778A] lymphocytes showed significantly lower chemotaxis towards chemokines and sphingosine 1-phosphate (S1P) than WT cells in vitro. The biggest migration defect was observed in response to S1P, which is essential for lymphocyte egress from secondary lymphoid organs. These results reveal a novel role of PKN1 in lymphocyte migration and localization.
敲入小鼠缺乏 PKN1 激酶活性,通过在催化结构域引入 T778A 点突变产生。PKN1[T778A] 突变小鼠成年后没有明显的外部异常,但外周血中的 T 和 B 淋巴细胞计数低于野生型 (WT) 小鼠。然而,在 PKN1[T778A] 小鼠的骨髓和胸腺中,T 和 B 细胞的发育显然是正常的,但是在淋巴结和脾脏中,突变小鼠的 T 和 B 细胞计数明显高于 WT 小鼠。在输注到 WT 受体后,与 EGFP 标记的 WT 供体淋巴细胞相比,EGFP 标记的 PKN1[T778A] 供体淋巴细胞在外周循环中的丰度明显较低,在受体小鼠的脾脏和淋巴结中的丰度明显较高,可能反映了以细胞自主方式在脾脏和淋巴结中隔离淋巴细胞。与 WT 细胞相比,PKN1[T778A] 淋巴细胞对趋化因子和鞘氨醇 1-磷酸 (S1P) 的体外趋化性明显降低。在对 S1P 的反应中观察到最大的迁移缺陷,S1P 是淋巴细胞从次级淋巴器官中迁出所必需的。这些结果揭示了 PKN1 在淋巴细胞迁移和定位中的新作用。