Fc Receptor Signaling to the Nucleus CARLOS ROSALES LEDEZMA - PowerPoint PPT Presentation

Fc Receptor Signaling to the Nucleus CARLOS ROSALES LEDEZMA Instituto de Investigaciones Biomédicas Universidad Nacional Autónoma de México

Receptors for Fc part of IgG Ab Macrophage Berken & Benacerraf J. Exp. Med. 1966; 123:119

Fc Receptor Functions IgG Fc γ R Phagocytosis ADCC Macrophage Respiratory Burst Degranulation Inflammatory Mediators IgE Gene Activation Cytokine Production Fc ε R Mast cell

Human IgG Fc Receptors Fc γ RI Fc γ RIIA Fc γ RIIB Fc γ RIIIA Fc γ RIIIB CD64 CD32 CD32 CD16 CD16 α -GPI γ 2 α α α γ 2 α γ 2 α β Macrophage Macrophage Mast Cell Macrophage Macrophage Neutrophil Neutrophil Mast Cell Basophil Neutrophil Neutrophil Mast Cell Basophils Mast Cell Eosinophil Eosinophil NK Cell Eosinophil Dendritic Cell Dendritic Cell Dendritic Cell Platelet B Cell ITAM Dendritic Cell Annu. Rev. Immunol . 1991; 9:457 ITIM Annu. Rev. Immunol . 1997; 15:203

FcR signaling (~1996) Fc receptor Membrane Y-kinase ? IL-1, IL-8 TNF I κ B NF- κ B Nucleus NF- κ B DNA

Ras signaling RTK Membrane GAP Ras Ras GDP GTP Sos Grb2 Raf MEK ERK Nucleus Elk-1 Gene DNA expression SRE

Assay for NF- κ B activation THP-1 Fc receptor Membrane Luciferase NF- κ B mRNA luc Gene I κ B NF- κ B

Dominant negative Ras does not inhibit NF- κ B activation Medium IIC Luciferase Activity 5 (Fold Increase) 4 3 2 1 0 pZIP Ras N17

Dominant negative Raf does not inhibit NF- κ B activation Medium IIC Luciferase Activity 6 (Fold Increase) 5 4 3 2 1 0 pCGN Raf 23-284

MEK is required for immune complex-mediated NF- κ B activation Medium IIC Luciferase Activity IIC + PD98059 4 (Fold Increase) 3 2 1 0

FcR signaling THP-1 Fc receptor Syk ? MEK ERK I κ B NF- κ B Nucleus NF- κ B Gene DNA Expression J. Biol. Chem. 1998; 273 : 27610

ERK activation requires PI 3-K -phospho-ERK

MEK activation depends on PI 3-K

PI 3-K is required for NF- κ B activation 4 3.5 Luciferase Activity 3 Medium 2.5 IIC 2 IIC + Wor 1.5 IIC + LY 1 0.5 0

FcR signaling THP-1 Fc receptor PI3K Syk MEK ERK I κ B NF- κ B Nucleus NF- κ B Gene DNA Expression J. Leukoc. Biol. 2001; 70 : 649

Neutrophil Fc Receptors Fc γ RIIA Fc γ RIIIB CD32 CD16 α -GPI α Phagocytosis Calcium Rise Calcium Rise Respiratory Burst Degranulation Actin assembly Respiratory Burst Integrin activation J. Biol. Chem. 1992; 267: 5265 Biochem. Biophys. Acta 1999; 1452: 46 J. Leukoc. Biol . 2005; 15: 203

Are FcRIIA and FcRIIIB Signaling Different? Fc γ RIIA Fc γ RIIIB ? PMN

Nuclear Factor Activation mAb Fc γ R Fc γ R I κ B NF- κ B NF- κ B PMN J. Immunol. Meth. 2007; 320: 104

Nuclear Factor Activation Fc γ R FITC NF- κ B NF- κ B NF- κ B FACS J. Immunol. Meth. 2007; 320: 104

3G8 NF- κ B Activation IV.3 --- 4 3 2 1 0 3.5 2.5 1.5 0.5 Nuclear Staining (Fold Increase)

Phagocytosis 5 4.5 4 3.5 Fold Increase 3 --- 2.5 IV.3 3G8 2 1.5 1 0.5 0 FcRIIA FcRIIIB

FcRIIIB signal to NF- κ B is independent of Syk and MEK Nuclear Staining (Fold Increase) 4 3.5 3 2.5 DMSO 2 Piceatannol PD98059 1.5 1 0.5 0 IV.3 3G8

Signaling to Nucleus in PMN Fc γ RIIA Fc γ RIIIB PMN PI3K Syk Nucleus MEK ERK I κ B NF- κ B NF- κ B

Signaling to Nucleus in PMN Fc γ RIIA Fc γ RIIIB PMN PI3K Syk ? Nucleus MEK P ERK P ERK I κ B NF- κ B NF- κ B

Nuclear phospho-ERK 3G8 IV.3 4 3 2 1 0 4.5 3.5 2.5 1.5 0.5 Nuclear Staining (Fold Increase)

Nuclear phospho-ERK F(ab’) 2 ---- IV.3 3G8 3G8 p-ERK ERK

Nuclear phopho-ERK is independent Syk, MEK, and PI 3-K Nuclear Staining (Fold Increase) 6 5 4 3 3G8 2 1 0 DMSO Piceatannol PD98059 Wortmannin

Signaling to Nucleus in PMN Fc γ RIIA Fc γ RIIIB PMN PI3K Syk Nucleus MEK ERK P ERK I κ B NF- κ B NF- κ B

Nuclear ERK does not change after Fc γ RIIIB crosslinking Nuclear ERK

MEK is not present in PMN nucleus Nuclear MEK

Nuclear phospho-Elk-1 Nuclear Staining (Fold Increase) 4.5 4 3.5 3 2.5 2 1.5 1 0.5 0 IV.3 3G8

Nuclear phospho-Elk-1 F(ab’) 2 ---- 3G8 3G8 p-Elk-1 ERK

Nuclear phopho-Elk-1 is independent Syk, MEK, and PI 3-K Nuclear Staining (Fold Increase) 6 5 4 3 3G8 2 1 0 DMSO Piceatannol PD98059 Wortmannin

Fc γ R activation of ERK in PMN cytoplasm Fc γ RIIA Fc γ RIIIB PMN PI3K Syk ? Nucleus MEK ERK P ERK I κ B NF- κ B NF- κ B

Fc γ RIIIB activates nuclear factors in PMN Fc γ RIIA Fc γ RIIIB PMN Nucleus ERK P I κ B NF- κ B Elk-1 P NF- κ B

Conclusions • Fc γ RIIA and Fc γ RIIIB signal differently in the PMN • Fc γ RIIA induces efficient phagocytosis • Fc γ RIIIB induces efficient ERK and Elk-1 activation • Fc γ RIIIB signal transduction does not involve Syk, MEK, PI 3-K, Src family kinases, or Tec family kinases

Acknowledgments • Gabriela Sánchez-Mejorada • Elsa Merit Reyes • Erick García • Nancy Mora • Alejandro Ortiz Stern • Georgina Nieto • Melissa Ruiz • Marco Hernández • Selma Rivas • Victor Salas