IL-2 signaling in human monocytes involves the phosphorylation and activation of p59hck.
The activating properties of IL-2 and the structure of the IL-2R on human monocytes are well characterized. However, relatively little is known about the biochemical mechanisms involved in IL-2 signal transduction in these cells. We investigated the role of protein tyrosine kinases (PTKs) in the activation of monocytes by IL-2. Incubation of monocytes with the PTK inhibitor herbimycin A (HA) resulted in the dose-dependent suppression of IL-2-induced monocyte tumoricidal activity. This inhibition was rather potent, as a concentration of HA as low as 0.5 microM caused a complete abrogation of cytolytic activity. Furthermore, HA markedly suppressed the ability of IL-2 to induce IL-1 beta, TNF-alpha, IL-6, and IL-8 mRNA expression and protein secretion by monocytes. Anti-phosphotyrosine immunoblotting demonstrated that IL-2 induced a rapid and time-dependent increase in tyrosine phosphorylation of several cellular proteins of molecular masses ranging from 35 to 180 kDa. Interestingly, IL-2 caused a significant up-regulation of the constitutive levels of hck PTK mRNA and protein relative to medium-treated cells as well as an increase in p59hck tyrosine phosphorylation. Finally, we demonstrated by in vitro kinase assay that the specific activity of p59hck PTK was also induced by IL-2 in monocytes. Thus, these data show that the activation of PTKs is required for the triggering of monocyte effector and secretory functions by IL-2 and strongly suggest that p59hck is a key participant in IL-2 signaling in human monocytes.
Bosco
M C
MC
Laboratory of Molecular Biology, Giannina Gaslini Institute, Genova Quarto, Italy.
Curiel
R E
RE
Zea
A H
AH
Malabarba
M G
MG
Ortaldo
J R
JR
Espinoza-Delgado
I
I
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D016227:Benzoquinones; D002478:Cells, Cultured; D016207:Cytokines / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D019073:HT29 Cells; D006801:Humans; D007249:Inflammation / Q000276:immunology / Q000517:prevention & control; D007376:Interleukin-2 / Q000494:pharmacology / Q000502:physiology*; D007399:Interphase / Q000276:immunology; D053613:Janus Kinase 1; D053616:Janus Kinase 3; D047029:Lactams, Macrocyclic; D008213:Lymphocyte Activation / Q000276:immunology*; D008262:Macrophage Activation / Q000187:drug effects / Q000276:immunology; D009000:Monocytes / Q000187:drug effects / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D051572:Proto-Oncogene Proteins c-hck; D011809:Quinones / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction / Q000276:immunology*; D015854:Up-Regulation / Q000235:genetics / Q000276:immunology; D019061:src-Family Kinases / Q000096:biosynthesis / Q000235:genetics
D000903:Antibiotics, Antineoplastic; D016227:Benzoquinones; D016207:Cytokines; D007376:Interleukin-2; D047029:Lactams, Macrocyclic; D011518:Proto-Oncogene Proteins; D011809:Quinones; D012333:RNA, Messenger; D017828:Rifabutin; D019000:Phosphotyrosine; C020754:herbimycin; D011505:Protein-Tyrosine Kinases; C496631:HCK protein, human; C507904:JAK1 protein, human; C507933:JAK3 protein, human; D053613:Janus Kinase 1; D053616:Janus Kinase 3; D051572:Proto-Oncogene Proteins c-hck; D019061:src-Family Kinases