Signal transduction of interleukin 2 in human natural killer cells: involvement of the p56lck tyrosine kinase.
Despite numerous reports, the role of the protein tyrosine kinase p56lck in IL-2 signal transduction has remained controversial. We show here, using IL-2-dependent human natural killer cell lines, that p56lck is regulated by IL-2 in two different ways: (1) IL-2 induces a rapid increase of p56lck kinase activity as assessed in vitro; and (2) following IL-2 stimulation, p56lck undergoes phosphorylation on serine residues that is reflected by a modification of its electrophoretic mobility in SDS-PAGE. Furthermore, dose response experiments, and blocking studies performed with anti-IL-2R alpha antibodies, indicated that binding of IL-2 to the IL-2R beta chain was sufficient to produce these modifications of p56lck. In contrast, activation of the CD2 pathway stimulated the kinase activity of p56lck, but did not induce a significant shift in NK cells, as opposed to T lymphocytes. Western blot analyses, and immunoprecipitations of cell lysates from 32P-preloaded NK cells demonstrated that seven major proteins are tyrosine phosphorylated in response to IL-2. These phosphoproteins, with apparent molecular weights of 190, 150, 120, 110, 85, 65 and 56, which may not all be p56lck substrates, undergo phosphorylation and dephosphorylation with different kinetics. Furthermore, pp120 was identified as rasGAP, by Western blot and immunoprecipitation experiments. rasGAP and some of its co-precipitating molecules become phosphorylated in response to IL-2, presumably by p56lck, which would thus provide a link between IL-2R and downstream events critical for NK cell proliferation and function.
Vitte-Mony
I
I
Institut National de la Santé et de la Recherche Médicale, Faculté de Pharmacie, Chatenay-Malabry, France.
Delespine-Carmagnat
M
M
Bertoglio
J H
JH
D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D015153:Blotting, Western; D018801:CD2 Antigens; D002460:Cell Line; D002999:Clone Cells; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D007376:Interleukin-2 / Q000502:physiology*; D007694:Killer Cells, Natural / Q000201:enzymology / Q000502:physiology*; D008213:Lymphocyte Activation / Q000502:physiology; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism / Q000502:physiology*; D011971:Receptors, Immunologic / Q000502:physiology; D015375:Receptors, Interleukin-2 / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D013601:T-Lymphocytes / Q000502:physiology
D000945:Antigens, Differentiation, T-Lymphocyte; D018801:CD2 Antigens; D007376:Interleukin-2; D011971:Receptors, Immunologic; D015375:Receptors, Interleukin-2; D011505:Protein-Tyrosine Kinases; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)