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Fc Receptor Signaling to the Nucleus
Instituto de Investigaciones BiomédicasUniversidad Nacional Autónoma de México
CARLOS ROSALES LEDEZMA
Receptors for Fc part of IgG
Berken & BenacerrafJ. Exp. Med.1966; 123:119
Macrophage
Ab
Fc Receptor Functions
Mast cell
IgE
FcεR
Macrophage
IgG
FcγR PhagocytosisADCCRespiratory BurstDegranulationInflammatory MediatorsGene ActivationCytokine Production
Human IgG Fc Receptors
FcγRI FcγRIIA FcγRIIB FcγRIIIA FcγRIIIB
CD64 CD32 CD32 CD16 CD16
γ2 α α α α α α-GPIγ2 γ2 β
MacrophageNeutrophilEosinophilDendritic Cell
MacrophageNeutrophilMast CellEosinophilPlateletDendritic Cell
MacrophageNeutrophilMast CellEosinophilDendritic CellB Cell
MacrophageMast CellBasophilsNK CellDendritic Cell
Mast CellBasophil
Neutrophil
ITAM
ITIMAnnu. Rev. Immunol. 1991; 9:457Annu. Rev. Immunol. 1997; 15:203
FcR signaling (~1996)
Nucleus
DNA
IL-1, IL-8TNF
?
Y-kinase
Fc receptor Membrane
NF-κB
NF-κBIκB
Ras signaling
RTK MembraneGAP
Grb2Sos
RasGDP
RasGTP
Raf
SRE
Nucleus
DNAGeneexpression
MEK
ERK
Elk-1
Assay for NF-κB activation
Luciferase
mRNAlucGene
Fc receptorTHP-1Membrane
NF-κBIκB
NF-κB
Dominant negative Ras does not inhibit NF-κB activation
0
1
2
3
4
5
pZIP Ras N17
Luci
fera
se A
ctiv
ity(F
old
Incr
ease
)
MediumIIC
Dominant negative Raf does not inhibit NF-κB activation
0
1
2
3
4
5
6
pCGN Raf 23-284
MediumIIC
Luci
fera
se A
ctiv
ity(F
old
Incr
ease
)
MEK is required for immune complex-mediated NF-κB activation
0
1
2
3
4
MediumIICIIC + PD98059
Luci
fera
se A
ctiv
ity(F
old
Incr
ease
)
FcR signaling
Nucleus
DNA
?
Fc receptor THP-1
NF-κB
NF-κBIκB
Syk
MEK
ERK
J. Biol. Chem. 1998; 273: 27610
GeneExpression
ERK activation requires PI 3-K
-phospho-ERK
MEK activation depends on PI 3-K
PI 3-K is required for NF-κB activation
0
0.5
1
1.5
2
2.5
3
3.5
4
Luci
fera
se A
ctiv
ity
MediumIICIIC + WorIIC + LY
FcR signaling
Nucleus
DNA
Fc receptor THP-1
NF-κB
NF-κBIκB
Syk
MEK
ERK
J. Leukoc. Biol. 2001; 70: 649
GeneExpression
PI3K
Neutrophil Fc ReceptorsFcγRIIA FcγRIIIB
CD32 CD16
α α-GPI
J. Leukoc. Biol. 2005; 15: 203
PhagocytosisCalcium RiseDegranulationRespiratory Burst
Calcium RiseRespiratory BurstActin assemblyIntegrin activation
J. Biol. Chem. 1992; 267: 5265Biochem. Biophys. Acta 1999; 1452: 46
Are FcRIIA and FcRIIIB Signaling Different?
FcγRIIIB
?
PMN
FcγRIIA
Nuclear Factor Activation
PMN
FcγR
mAb
NF-κB
IκB
FcγR
NF-κB
J. Immunol. Meth. 2007; 320: 104
Nuclear Factor Activation
FcγR
NF-κB
NF-κB NF-κB
FITC
FACSJ. Immunol. Meth. 2007; 320: 104
NF-κB Activation
00.5
11.5
22.5
33.5
4
--- IV.3 3G8
Nuc
lear
Sta
inin
g (F
old
Incr
ease
)
Phagocytosis
00.5
11.5
22.5
33.5
44.5
5
FcRIIA FcRIIIB
Fold
Incr
ease
---IV.33G8
FcRIIIB signal to NF-κB is independent of Syk and MEK
00.5
11.5
22.5
33.5
4
IV.3 3G8
Nuc
lear
Sta
inin
g (F
old
Incr
ease
)
DMSOPiceatannolPD98059
Signaling to Nucleus in PMNFcγRIIA FcγRIIIB
SykPI3K
MEK
ERK
NF-κB
NF-κBIκB
PMN
Nucleus
Signaling to Nucleus in PMNFcγRIIA FcγRIIIB
SykPI3K
MEK
ERK
NF-κB
NF-κBIκB
PMN
Nucleus
ERKP
?
P
Nuclear phospho-ERK
00.5
11.5
22.5
33.5
44.5
IV.3 3G8
Nuc
lear
Sta
inin
g (F
old
Incr
ease
)
Nuclear phospho-ERK
ERK
---- 3G8 F(ab’)2 3G8IV.3
p-ERK
Nuclear phopho-ERK is independent Syk, MEK, and PI 3-K
0
1
2
3
4
5
6
DMSO Piceatannol PD98059 Wortmannin
Nuc
lear
Sta
inin
g (F
old
Incr
ease
)
3G8
Signaling to Nucleus in PMNFcγRIIA FcγRIIIB
SykPI3K
MEK
ERK
NF-κB
NF-κBIκB
PMN
Nucleus
ERKP
Nuclear ERK does not change after FcγRIIIB crosslinking
Nuclear ERK
MEK is not present in PMN nucleus
Nuclear MEK
Nuclear phospho-Elk-1
00.5
11.5
22.5
33.5
44.5
IV.3 3G8
Nuc
lear
Sta
inin
g (F
old
Incr
ease
)
Nuclear phospho-Elk-1
p-Elk-1
ERK
---- 3G8 F(ab’)2 3G8
Nuclear phopho-Elk-1 is independent Syk, MEK, and PI 3-K
0
1
2
3
4
5
6
DMSO Piceatannol PD98059 Wortmannin
Nuc
lear
Sta
inin
g (F
old
Incr
ease
)
3G8
FcγR activation of ERK in PMN cytoplasm
FcγRIIA FcγRIIIB
SykPI3K
MEK
ERK
NF-κBNF-κBIκB
PMN
Nucleus
ERK P
?
FcγRIIIB activates nuclear factors in PMN
FcγRIIA FcγRIIIB
NF-κBNF-κBIκB
PMN
Nucleus
ERK P
Elk-1 P
Conclusions
• FcγRIIA and FcγRIIIB signal differently in the PMN
• FcγRIIA induces efficient phagocytosis
• FcγRIIIB induces efficient ERK and Elk-1 activation
• FcγRIIIB 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