title: Essential role of the adaptor protein Nck1 in Jurkat T cell activation and function.
abstract: The non-catalytic region of tyrosine kinase (Nck) is proposed to play an essential role in T cell activation. However, evidence based on functional and biochemical studies has brought into question the critical function of Nck. Therefore, the aim of the present work was to investigate the role of Nck in T cell activation. To study this, the human Jurkat T cell line was used as a model for human T lymphocytes. The short interfering (si) RNA targeting Nck1 gene was used with electroporation to knock-down Nck1 protein expression in Jurkat T cells. Primary human CD4 T cells were also transfected with the siRNA of Nck1. The results showed that decreased Nck1 protein expression did not affect the apoptosis of the transfected Jurkat T cells compared with control siRNA-transfected cells and non-transfected cells. Upon CD3ε/CD28 stimulation, knock-down of Nck1 in Jurkat T cells caused a decrease in CD69 expression and in interleukin (IL)-2 secretion. Similarly, knock-down of Nck1 in primary CD4 T cells also caused decreased CD69 expression. However, no significant alterations of CD69 and IL-2 expression were found upon phytohaemagglutinin (PHA)/phorbol myristate acetate (PMA) stimulation. Knock-down of Nck1 had no effect on the proliferation of Jurkat T cells stimulated with either PHA or anti-T cell receptor (TCR) monoclonal antibody (C305). The reduced Nck1 expression in Jurkat cells was also associated with a reduced phosphorylation of extracellular regulated kinase (Erk)1 and Erk2 proteins upon CD3ε/CD28 stimulation. In conclusion, the decreased Nck1 protein in Jurkat T cells resulted in an impairment of TCR-CD3-mediated activation involving a defective Erk phosphorylation pathway.
authors: Department of Preventive Dentistry, Faculty of Dentistry, Naresuan University, Phitsanulok, Thailand.
authors: I
authors: I
authors: Yiemwattana
authors: J
authors: J
authors: Ngoenkam
authors: P
authors: P
authors: Paensuwan
authors: R
authors: R
authors: Kriangkrai
authors: B
authors: B
authors: Chuenjitkuntaworn
authors: S
authors: S
authors: Pongcharoen
mesh_terms: D056704:Adaptive Immunity / Q000276:immunology; D048868:Adaptor Proteins, Signal Transducing / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis / Q000235:genetics; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D018106:CD28 Antigens / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D018274:Electroporation; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis / Q000235:genetics; D019169:Jurkat Cells / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D037181:Lectins, C-Type / Q000096:biosynthesis / Q000235:genetics; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D015513:Oncogene Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D010766:Phosphorylation; D010835:Phytohemagglutinins / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000276:immunology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D000099192:CD69 Antigens
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D018106:CD28 Antigens; D007375:Interleukin-1; D037181:Lectins, C-Type; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D015513:Oncogene Proteins; D010835:Phytohemagglutinins; D034741:RNA, Small Interfering; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D013755:Tetradecanoylphorbol Acetate; D000099192:CD69 Antigens; C078332:Nck protein; C535150:MAPK1 protein, human