Critical role of protein kinase C ϵ for lipopolysaccharide‐induced IL‐12 synthesis in monocyte‐derived dendritic cells

In the present study we have investigated the potential involvement of protein kinase C (PKC) in the maturation of human dendritic cells (DC) by bacterial lipopolysaccharide (LPS). LPS stimulation of DC derived from human monocytes resulted in PKC phosphorylation. Inhibition of PKC activation using bisindolylmaleimide (Bis), a pan‐PKC inhibitor, was associated with a dose‐dependent decrease of LPS‐induced IL‐12 production. In contrast, up‐regulation of MHC class II, CD80 and CD86 was not altered. Consistent with the diminished IL‐12 synthesis, DC stimulated with LPS in presence of Bis were deficient in the induction of IFN‐γ production by allogeneic CD4+ T cells. Furthermore, we found that PKC inhibition impaired LPS‐induced IκB‐α degradation and subsequent nuclear factor (NF)‐κB activation in DC. LPS resulted in the phosphorylation of conventional α/β and novel ϵ PKC isoforms in DC. Inhibition of LPS‐induced PKC activity using pseudosubstrate peptides specific for PKC isoforms established that PKC ϵ but not PKC α/β was involved in the production of IL‐12 and TNF‐α. Overall, these data provide evidence thatPKC inhibition impairs LPS signaling in DC and identify PKC ϵ as a potential target for the inhibition of Toll‐like receptor‐4‐mediated, IL‐12‐dependent Th1 type responses.

[1]  D. Harlan,et al.  Evidence for Distinct Intracellular Signaling Pathways in CD34+ Progenitor to Dendritic Cell Differentiation from a Human Cell Line Model , 1999, The Journal of Immunology.

[2]  Ye Zheng,et al.  Dendritic Cell Development and Survival Require Distinct NF-κB Subunits , 2002 .

[3]  Ye Zheng,et al.  Dendritic cell development and survival require distinct NF-kappaB subunits. , 2002, Immunity.

[4]  D. Pennington,et al.  Protein Kinase C (cid:2) Is Required for Macrophage Activation and Defense Against Bacterial Infection , 2001 .

[5]  Meredith O'Keeffe,et al.  C-Rel Regulates Interleukin 12 P70 Expression in Cd8+ Dendritic Cells by Specifically Inducing p35 Gene Transcription , 2001, The Journal of experimental medicine.

[6]  F. Re,et al.  Toll-like Receptor 2 (TLR2) and TLR4 Differentially Activate Human Dendritic Cells* , 2001, The Journal of Biological Chemistry.

[7]  N. Kadowaki,et al.  Subsets of Human Dendritic Cell Precursors Express Different Toll-like Receptors and Respond to Different Microbial Antigens , 2001, The Journal of experimental medicine.

[8]  N. Verdaguer,et al.  Structure of the C2 domain from novel protein kinase Cepsilon. A membrane binding model for Ca(2+)-independent C2 domains. , 2001, Journal of molecular biology.

[9]  V. Soumelis,et al.  Dendritic cell lineage, plasticity and cross-regulation , 2001, Nature Immunology.

[10]  S. Akira,et al.  Endotoxin-Induced Maturation of MyD88-Deficient Dendritic Cells1 , 2001, The Journal of Immunology.

[11]  V. Kindler,et al.  A Critical Role for p38 Mitogen-Activated Protein Kinase in the Maturation of Human Blood-Derived Dendritic Cells Induced by Lipopolysaccharide, TNF-α, and Contact Sensitizers , 2001, The Journal of Immunology.

[12]  J. Vekemans,et al.  Deficient IL-12(p35) Gene Expression by Dendritic Cells Derived from Neonatal Monocytes1 , 2001, The Journal of Immunology.

[13]  D. Charon,et al.  The Bisindolylmaleimide GF 109203X Is a Potent and Selective Inhibitor of Protein Kinase C* , 2001 .

[14]  C. Reis e Sousa,et al.  Dendritic cells as sensors of infection. , 2001, Immunity.

[15]  A. Hoffmann,et al.  Nucleosome remodeling at the IL-12 p40 promoter is a TLR-dependent, Rel-independent event , 2001, Nature Immunology.

[16]  A. Hoffmann,et al.  Selective requirement for c-Rel during IL-12 P40 gene induction in macrophages. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[17]  C. Janeway,et al.  The Toll receptor family and microbial recognition. , 2000, Trends in microbiology.

[18]  Asim Khwaja,et al.  The PI3 kinase, p38 SAP kinase, and NF-κB signal transduction pathways are involved in the survival and maturation of lipopolysaccharide-stimulated human monocyte–derived dendritic cells , 2000 .

[19]  E. Schaeffer,et al.  PKC-θ is required for TCR-induced NF-κB activation in mature but not immature T lymphocytes , 2000, Nature.

[20]  E. Kontny,et al.  Rottlerin, a PKC isozyme‐selective inhibitor, affects signaling events and cytokine production in human monocytes , 2000, Journal of leukocyte biology.

[21]  E. Bröcker,et al.  Differential expression of Rel/NF-κB and octamer factors is a hallmark of the generation and maturation of dendritic cells , 2000 .

[22]  E. Schaeffer,et al.  PKC-theta is required for TCR-induced NF-kappaB activation in mature but not immature T lymphocytes. , 2000, Nature.

[23]  C Caux,et al.  Immunobiology of dendritic cells. , 2000, Annual review of immunology.

[24]  K M Ardeshna,et al.  The PI3 kinase, p38 SAP kinase, and NF-kappaB signal transduction pathways are involved in the survival and maturation of lipopolysaccharide-stimulated human monocyte-derived dendritic cells. , 2000, Blood.

[25]  M. Karin,et al.  Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. , 2000, Annual review of immunology.

[26]  E. Bröcker,et al.  Differential expression of Rel/NF-kappaB and octamer factors is a hallmark of the generation and maturation of dendritic cells. , 2000, Blood.

[27]  S. Smale,et al.  Rapid and selective remodeling of a positioned nucleosome during the induction of IL-12 p40 transcription. , 1999, Immunity.

[28]  W. Maśliński,et al.  Protein kinase c-dependent pathway is critical for the production of pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6). , 1999, Cytokine.

[29]  D. Ballard,et al.  IκB Kinase Complex Is an Intracellular Target for Endotoxic Lipopolysaccharide in Human Monocytic Cells , 1999 .

[30]  S. Morony,et al.  TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling. , 1999, Genes & development.

[31]  Katsuaki Sato,et al.  Extracellular signal-regulated kinase, stress-activated protein kinase/c-Jun N-terminal kinase, and p38mapk are involved in IL-10-mediated selective repression of TNF-alpha-induced activation and maturation of human peripheral blood monocyte-derived dendritic cells. , 1999, Journal of immunology.

[32]  D. Ballard,et al.  IkappaB kinase complex is an intracellular target for endotoxic lipopolysaccharide in human monocytic cells. , 1999, Blood.

[33]  Z. Wang,et al.  Bisindolylmaleimide VIII facilitates Fas-mediated apoptosis and inhibits T cell-mediated autoimmune diseases , 1999, Nature Medicine.

[34]  P. Ricciardi-Castagnoli,et al.  Brief Definitive Report Dendritic Cell Survival and Maturation Are Regulated by Different Signaling Pathways , 2022 .

[35]  A. Manning,et al.  Role of IKK1 and IKK2 in Lipopolysaccharide Signaling in Human Monocytic Cells* , 1998, The Journal of Biological Chemistry.

[36]  C. Janeway,et al.  MyD88 is an adaptor protein in the hToll/IL-1 receptor family signaling pathways. , 1998, Molecular cell.

[37]  P. Parker,et al.  The extended protein kinase C superfamily. , 1998, The Biochemical journal.

[38]  D. Harlan,et al.  Phorbol esters induce differentiation of human CD34+ hemopoietic progenitors to dendritic cells: evidence for protein kinase C-mediated signaling. , 1998, Journal of immunology.

[39]  M J May,et al.  NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. , 1998, Annual review of immunology.

[40]  D. Mochly‐Rosen,et al.  Anchoring proteins for protein kinase C: a means for isozyme selectivity. , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[41]  A. Sher,et al.  In Vivo Microbial Stimulation Induces Rapid CD40 Ligand–independent Production of Interleukin 12 by Dendritic Cells and their Redistribution to T Cell Areas , 1997, The Journal of experimental medicine.

[42]  B. Boyan,et al.  Bacterial lipopolysaccharide induces early and late activation of protein kinase C in inflammatory macrophages by selective activation of PKC-epsilon. , 1997, Biochemical and Biophysical Research Communications - BBRC.

[43]  S. Smale,et al.  Multiple control elements mediate activation of the murine and human interleukin 12 p40 promoters: evidence of functional synergy between C/EBP and Rel proteins , 1997, Molecular and cellular biology.

[44]  Johann Hofmann,et al.  The potential for isoenzyme‐selective modulation of protein kinase C , 1997, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[45]  V. Verhasselt,et al.  Interleukin‐10 prevents the generation of dendritic cells from human peripheral blood mononuclear cells cultured with interleukin‐4 and granulocyte/ macrophage‐colony‐stimulating factor , 1997, European journal of immunology.

[46]  F. Sallusto,et al.  Origin, maturation and antigen presenting function of dendritic cells. , 1997, Current opinion in immunology.

[47]  P. Parker,et al.  Up-Regulation of Protein Kinase C-ε Promotes the Expression of Cytokine-inducible Nitric Oxide Synthase in RAW 264.7 Cells* , 1996, The Journal of Biological Chemistry.

[48]  D. Mochly‐Rosen,et al.  A Protein Kinase C Translocation Inhibitor as an Isozyme-selective Antagonist of Cardiac Function* , 1996, The Journal of Biological Chemistry.

[49]  A. Tarakhovsky,et al.  Immunodeficiency in Protein Kinase Cβ-Deficient Mice , 1996, Science.

[50]  P. Bergman,et al.  Partial Reversal of Multidrug Resistance in Human Breast Cancer Cells by an N-Myristoylated Protein Kinase C- Pseudosubstrate Peptide (*) , 1996, The Journal of Biological Chemistry.

[51]  G. Schuler,et al.  Generation of mature dendritic cells from human blood. An improved method with special regard to clinical applicability. , 1997, Advances in experimental medicine and biology.

[52]  A. Tarakhovsky,et al.  Immunodeficiency in protein kinase cbeta-deficient mice. , 1996, Science.

[53]  K. Murphy,et al.  Regulation of interleukin 12 p40 expression through an NF-kappa B half-site , 1995, Molecular and cellular biology.

[54]  D. Lo,et al.  Expression of relB is required for the development of thymic medulla and dendritic cells , 1995, Nature.

[55]  G. Trinchieri.  Interleukin-12: a proinflammatory cytokine with immunoregulatory functions that bridge innate resistance and antigen-specific adaptive immunity. , 1995, Annual review of immunology.

[56]  T. Yoshida,et al.  LPS induces selective translocation of protein kinase C-beta in LPS-responsive mouse macrophages, but not in LPS-nonresponsive mouse macrophages. , 1994, Journal of immunology.

[57]  N. Reiner,et al.  CD14-dependent activation of protein kinase C and mitogen-activated protein kinases (p42 and p44) in human monocytes treated with bacterial lipopolysaccharide. , 1994, Journal of immunology.

[58]  C. Janeway,et al.  The immune system evolved to discriminate infectious nonself from noninfectious self. , 1992, Immunology today.

[59]  H. Coste,et al.  The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. , 1991, The Journal of biological chemistry.

[60]  G. Nabel,et al.  Tumor necrosis factor alpha and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor kappa B. , 1989, Proceedings of the National Academy of Sciences of the United States of America.