CARLOS M. MINUTTI

Tissue-specific signals that modulate IL-4Rα–mediated macrophage activation in lung repair

XIIè CONGRÉS Societat Catalana d’Immunologia (SCI)

16.11.2018

Les macrophages font tout!!!

1884 2018

Élie Metchnikoff

Ruslan Medzhitov

Judi AllenMiriam Merad

Paradigm of macrophage activation

Sources of macrophage diversity: Origin and Niche

from Guilliams & Scott. Nature Reviews Immunology 2017

The alveoli provide an unique niche for macrophages

SP-A is an abundant component of the humoral immune defense in the lungs

SP-A is a member of the family of collectins

adapted from Sanchez-Barbero et al. Biochem. J. 2007

SP-A prevents viral and bacterial infections

adapted from Wright. Nature Immunology 2005

SP-A is also immuno-modulatory

adapted from Wright. Nature Immunology 2005

Does SP-A modulates activation of alveolar macrophages?

?

Mφ(IFN-γ + LPS) Mφ(IL-4)

?

SP-A attenuates classical activation of alveolar macrophages

Minutti et al. The Journal of Immunology 2016

Does SP-A modulates M(IL-4)?

?

Mφ(IFN-γ + LPS) Mφ(IL-4)

Is SP-A a tissue-specific factor?

Alveolar Peritoneal

CD11c+SiglecF+

CD11b+F480+

+ IL-4

?

Does another defense collagen, C1q, modulates IL-4 effects on alveolar and peritoneal Mf?

Alveolar Peritoneal

CD11c+SiglecF+

CD11b+F480+

+ IL-4

??

?

Experimental approach

Platemacrophages

Collagens and/or IL-4

Harvest

1.5 hours 48 hours

Proliferation Alternative activation

∑ BrdU∑ Ki67

∑ RELMα∑ Ym1

BrdU

SP-A and C1q are local tissue-specific amplifiers of IL-4Rα–mediated macrophage activation

Alveolar Mϕ Peritoneal Mϕ

Prol

ifera

tion

Activ

atio

n

Minutti et al. Science 2017

The effects of SP-A and C1q are similar in vivo

A-Mϕ P-Mϕ

Prol

ifera

tion

Activ

atio

n

A-Mϕ P-Mϕ

What is the receptor for SP-A and C1q?

Alveolar Mϕ Peritoneal MϕPr

olife

ratio

nAc

tivat

ion

Where is Myo18A expressed?

What is the effect of C1q in the liver?

Proliferation Activationin

viv

oin

vitr

o

IL-4 also drives local production of SP-A and C1q

LiverLung Peritoneum

What is the physiological relevance of SP-A during a model of type 2 immunity?

Day 6

Chen et al.; Nat Medicine 18,2012

Alveolar macrophages stimulatedwith IL-4/IL-13 are essential tomediate lung repair .

ProliferationAlternative activation

SP-A

?

? ? ?

?

Does SP-A influencethe ability of aMfs to

respond to IL-4?

D0: Subcutaneous inoculation of L3 larvae

D1 D4 D6

Migration

Type 2 macrophage activation decreases and lung damage increases in the absence of SP-A

What is the physiological relevance of C1q in a model of liver damage

from Blériot, et al. Immunity. 2015

WT

C1qa(-/-)

C57BL/6 mice

C1q-deficient mice showed increased liver damage after infection with Listeria monocytogenes

What is the physiological relevance of C1q during a model of peritoneal fibrosis?

from Wang, et al. Int. J. Mol. Sci. 2013

WT

C1qa(-/-)

C57BL/6 mice

Day 0 Day 29Sample

collection

2 4 6 8 10 12 14 16 18 20 22 24 26 28

Dianeal PD-4 administration (i. p.)

Peritoneal dialysis

Peritoneal fibrosis induced by a lactate dialysate is reduced in C1q-deficient mice

Summary

Minutti et al. Science, 2017

Bosurgi et al. Science, 2017

Recognition of apoptotic cells also enhances tissue repair function of macrophages

SP-A increases the phagocytosis of dying cells

adapted from Ravichandran, Journal of Experimental Medicine, 2010

What about the molecular mechanism?

SP-A amplifies IL-4 signaling on alveolar macrophages

P-STAT6 P-Akt

Myosin18A mediates SP-A reinforcement of IL-4-dependent effects

P-STAT6 P-Akt

ModeradorNotas de la presentaciónSTAT6 - Tyr641 Akt - Ser47

Known SP-A’s signaling pathways

PI3KBeharka, et al, JI 2002

P-AktP-PKCζMoulakakis, et al, JI 2007

PI3K inhibitor blocks SP-A effects on p-Akt and p-PKCz

PI3K

P-AktP-PKCζ P-Akt P-PKCζ

ModeradorNotas de la presentaciónPKC - Thr410/403 Akt - Ser47

PI3K inhibitor blocks SP-A actions on IL-4 effects

PI3K

P-AktP-PKCζ

Activation Proliferation

Akt activation controls SP-A and IL-4-mediated macrophage proliferation

PI3K

P-AktP-PKCζ

Activation Proliferation

PKCζ activation mediates SP-A enhancement of IL-4-induced activation of alveolar macrophages

PI3K

P-AktP-PKCζ

Activation Proliferation

Conclusions

IL-4 SP-A

Acknowledgements

Cristina Casals’s lab and CIBERES

Belén García-Fojeda (Mad)Raquel Guillamat-Prats (BCN)Antonio Artigas (BCN)

Judi Allen’s lab

Lucy Jackson-Jones (Ed)Tara Sutherland (Ed)Nicola Logan (Ed)Emma Rinqvist (Ed)

CollaboratorsJohanna Knipper (UK)David Ferenbach (UK) Cordula Stamme (DE)Zissis Chroneos (US)

Dietmar Zaiss’s lab

ES UK

UNIVERSIDAD COMPLUTENSE

Número de diapositiva 1Les macrophages font tout!!!Paradigm of macrophage activationSources of macrophage diversity: Origin and NicheThe alveoli provide an unique niche for macrophagesSP-A is an abundant component of the humoral immune defense in the lungsSP-A is a member of the family of collectinsSP-A prevents viral and bacterial infectionsSP-A is also immuno-modulatoryDoes SP-A modulates activation of alveolar macrophages?SP-A attenuates classical activation of alveolar macrophagesDoes SP-A modulates M(IL-4)?Is SP-A a tissue-specific factor? Número de diapositiva 14SP-A and C1q are local tissue-specific amplifiers of IL-4Rα–mediated macrophage activation The effects of SP-A and C1q are similar in vivoWhat is the receptor for SP-A and C1q?Where is Myo18A expressed?What is the effect of C1q in the liver?IL-4 also drives local production of SP-A and C1qWhat is the physiological relevance of SP-A during a model of type 2 immunity?Type 2 macrophage activation decreases and lung damage increases in the absence of SP-AWhat is the physiological relevance of C1q in a model of liver damageC1q-deficient mice showed increased liver damage after infection with Listeria monocytogenesWhat is the physiological relevance of C1q during a model of peritoneal fibrosis?Peritoneal fibrosis induced by a lactate dialysate is reduced in C1q-deficient miceSummaryNúmero de diapositiva 28SP-A increases the phagocytosis of dying cells What about the molecular mechanism?SP-A amplifies IL-4 signaling on alveolar macrophages Myosin18A mediates SP-A reinforcement of IL-4-dependent effects Known SP-A’s signaling pathwaysPI3K inhibitor blocks SP-A effects on p-Akt and p-PKCPI3K inhibitor blocks SP-A actions on IL-4 effectsAkt activation controls SP-A and IL-4-mediated macrophage proliferationPKCζ activation mediates SP-A enhancement of IL-4-induced activation of alveolar macrophagesConclusionsAcknowledgements