Institute of Cell Genetics, Medical University Innsbruck, Innsbruck, Austria.
Apoptosis, Cancer, and Development Laboratory, Equipe labellisée 'La Ligue', LabEx DEVweCAN, Centre de Recherche en Cancérologie de Lyon, INSERM U1052-CNRS UMR5286, Centre Léon Bérard, Université de Lyon, Université Claude Bernard Lyon1, 69008, Lyon, France.
Cell Commun Signal. 2022 Apr 19;20(1):54. doi: 10.1186/s12964-022-00864-w.
The Protein kinase D3 (PKD3) has been implicated in signal transduction downstream of the T cell receptor (TCR). However, its role for the activation of primary T lymphocytes has not been elucidated so far.
Expression of PKD isoforms in primary murine T cells was determined by RT-PCR and SDS-Page. A germline PKD3-knockout mouse line was analyzed for its immune response to OVA/alum intraperitoneal immunization. Phenotyping of the T cell compartment ex vivo as well as upon stimulation in vitro was performed by flow cytometry. Additionally, cytokine expression was assessed by flow cytometry, RT-PCR and Luminex technology.
PKD expression in T cells is modulated by TCR stimulation, leading to a rapid down-regulation on mRNA and on protein level. PKD3-deficient mice respond to immunization with enhanced T follicular helper cell generation. Furthermore, peripheral PKD3-deficient CD4 T cells express more interleukin-2 than wild type CD4 T cells upon TCR stimulation ex vivo. However, purified naïve CD4 T cells do not differ in their phenotype upon differentiation in vitro from wild type T cells. Moreover, we observed a shift towards an effector/memory phenotype of splenic T cells at steady state, which might explain the contradictory results obtained with pan-T cells ex vivo and naïve-sorted T cells.
While PKD3-deficiency in vivo in mice leads to a skewing of the T cell compartment towards a more activated phenotype, this kinase seems to be dispensable for naïve CD4 T cell differentiation in vitro. Video Abstract.
蛋白激酶 D3（PKD3）已被牵涉到 T 细胞受体（TCR）下游的信号转导中。然而，其对于原代 T 淋巴细胞的激活作用尚未阐明。
通过 RT-PCR 和 SDS-PAGE 测定原代鼠 T 细胞中 PKD 同工型的表达。分析了一个胚系 PKD3 敲除小鼠系对 OVA/明矾腹腔免疫的免疫反应。通过流式细胞术对 T 细胞区室进行体外表型分析以及体外刺激。此外，通过流式细胞术、RT-PCR 和 Luminex 技术评估细胞因子表达。
T 细胞中 PKD 的表达受 TCR 刺激调节，导致 mRNA 和蛋白水平的快速下调。PKD3 缺陷小鼠对免疫接种的反应增强了滤泡辅助性 T 细胞的生成。此外，外周 PKD3 缺陷的 CD4 T 细胞在 TCR 刺激下表达比野生型 CD4 T 细胞更多的白细胞介素 2。然而，在体外分化时，纯化的幼稚 CD4 T 细胞在表型上与野生型 T 细胞没有差异。此外，我们观察到在稳定状态下脾脏 T 细胞向效应/记忆表型的转变，这可能解释了在体外使用 pan-T 细胞和幼稚分选 T 细胞获得的矛盾结果。
虽然体内 PKD3 缺陷导致 T 细胞区室向更活跃的表型倾斜，但这种激酶对于体外幼稚 CD4 T 细胞分化似乎是可有可无的。
Zotero 插件
