Endothelial nitric oxide synthase regulates T cell receptor signaling at the immunological synapse.
The role of nitric oxide (NO) in T cells remains controversial, and the origin and localization of endogenous NO and whether it regulates lymphocyte activation are unclear. We show here that, within minutes of binding to antigen, T cells produce NO via endothelial nitric oxide synthase (eNOS). This process required increased intracellular Ca2+ and phosphoinositide3-kinase activity. By using an eNOS-green fluorescent fusion protein and fluorescent probes to detect NO, we show that eNOS translocates with the Golgi apparatus to the immune synapse of T helper cells engaged with antigen-presenting cells (APC), where it was fully activated. Overexpression of eNOS prevented the central coalescence of CD3 at the T cell-APC contact site, which was accompanied by increased phosphorylation of CD3zeta chain, ZAP-70, and extracellular signal-regulated kinases and increased IFN-gamma synthesis, but reduced production of IL-2. Therefore, eNOS-derived NO selectively potentiates T cell receptor signaling to antigen at the immunological synapse.
Ibiza
Sales
S
Unidad Mixta: Centro Nacional de Investigaciones Cardiovasculares (CNIC), Universitat de Valencia, Avenida Blasco Ibáñez 15-17, E-46010 Valencia, Spain.
Víctor
Víctor M
VM
Unidad Mixta: Centro Nacional de Investigaciones Cardiovasculares (CNIC), Universitat de Valencia, Avenida Blasco Ibáñez 15-17, E-46010 Valencia, Spain.
Boscá
Irene
I
Unidad Mixta: Centro Nacional de Investigaciones Cardiovasculares (CNIC), Universitat de Valencia, Avenida Blasco Ibáñez 15-17, E-46010 Valencia, Spain.
Ortega
Angel
A
Unidad Central de Investigación, Universitat de Valencia, E-46010 Valencia, Spain.
Urzainqui
Ana
A
Servicio de Inmunología, Hospital de la Princesa, Universidad Autónoma de Madrid, E-28006 Madrid, Spain.
O'Connor
José E
JE
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universitat de Valencia, E-46010 Valencia, Spain.
Sánchez-Madrid
Francisco
F
Servicio de Inmunología, Hospital de la Princesa, Universidad Autónoma de Madrid, E-28006 Madrid, Spain.
Esplugues
Juan V
JV
Departamento de Farmacología, Facultad de Medicina, Universitat de Valencia, E-46010 Valencia, Spain.
Serrador
Juan M
JM
Unidad Mixta: Centro Nacional de Investigaciones Cardiovasculares (CNIC), Universitat de Valencia, Avenida Blasco Ibáñez 15-17, E-46010 Valencia, Spain. Electronic address: jmserrador@cnic.es.
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D000941:Antigens / Q000494:pharmacology; D017252:CD3 Complex / Q000032:analysis / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D008817:Mice, Mutant Strains; D009569:Nitric Oxide / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000819:agonists / Q000378:metabolism*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000201:enzymology* / Q000276:immunology
D000941:Antigens; D017252:CD3 Complex; D007376:Interleukin-2; D011948:Receptors, Antigen, T-Cell; D009569:Nitric Oxide; D007371:Interferon-gamma; D052250:Nitric Oxide Synthase Type III; D019869:Phosphatidylinositol 3-Kinases; D002118:Calcium