1

Director, Lydia Becker Institute ofImmunology and Inflammation

University of Manchester, UK.Tracy.hussell@manchester.ac.uk

Decision making in lung immunity

Professor Tracy Husselltracy.hussell@manchester.ac.uk

• Tissue complexity• Homeostasis• Normal processes that do not stop

Multiple factors affecting macrophage inflammatory tone

9.4

1.4

0.7

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ApoeF13a1H2-EaCd63Prkcb1H2-Ab1CkbMfge8AA467197H2-DMb1Fcgr2bEcm1LOC641240Serpina3gSlc6a8Ppap2bCcr5Lat2Lyz1S100a6AnpepNav1Cd74Iap

Nusap1Igf2bp2Ppap2aTk1AsnsLrig32310046K01RikP2ry13Wfdc10Acot1Prc1Trem1RhofCidecMyo6LipfWtipTnfaip8Nrn1Vstm2aAcaa1bKrt79Nt5eCar4

0 400 3000

No difference in 6526 probes

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No difference in 43 miRNAs

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9324324-5p484328125a-5p200b450-5p322503

0 1 3

11

Fol

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A B

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Ch 15:

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50401332 50407161 50407432

5740bp 201bp

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Fluorescence intensity

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99bp 94bp

let-7c294bp 84bp

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Nusap1Igf2bp2Ppap2aTk1AsnsLrig32310046K01RikP2ry13Wfdc10Acot1Prc1Trem1RhofCidecMyo6LipfWtipTnfaip8Nrn1Vstm2aAcaa1bKrt79Nt5eCar4

0 400 3000

No difference in 6526 probes

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Low

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No difference in 43 miRNAs

let-7b1851521629c42522129b27a378

9324324-5p484328125a-5p200b450-5p322503

0 1 3

11

Fol

d ch

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0.2

0.8

1

A B

Fol

d ch

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Ch 15:

Ch X:70bp

50401332 50407161 50407432

5740bp 201bp

C

Fold change

Fluorescence intensity

D

E

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85537749

99bp 94bp

let-7c294bp 84bp

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Res

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Blind folding airway macrophages: miRNA modifications

Dr Alexander Godlee

The impact of the extracellular matrix on inflammation. Lydia SorokinNature Reviews Immunology volume 10, pages 712–723 (2010)000

collagen type XVIII alpha 1 chain Nephronectin: binds

integrin a8b1

The instructive extracellular matrix of the lung: basic composition and alterations in chronic lung diseaseGerald Burgstaller, Bettina Oehrle, Michael Gerckens, Eric S. White, Herbert B. Schiller, Oliver Eickelberg. European Respiratory Journal 2017, 50: 1601805; DOI: 10.1183/13993003.01805-2016

Fibrillar collagen Fibronectin

X-linked fibrillar

collagen

Basal membrane Hyaluronan

Myofibroblast Tenascin C IntegrinsFibroblast

Hyaluronan is a major constituent of the inflamed lung

Why does matrix persist? Production versus degradation

Digestion of HA restores lung function

Hyaluronidase at d15 after influenza

Mechanosensing podosomes exert a protrusive force that senses environment stiffness

F-actin-rich cone-shaped structures ~600 nm height and submicron diameter,

Located at the basal face of cells, perpendicular to, and in contact with, the substratum via integrins at the podosome ring

Typical lifespan of 10 min, with an actin turnover of ~30 s

Fujimori T, Grabiec AM, Kaur M, Bell TJ, Fujino N, Cook PC, Svedberg FR, MacDonald AS, Maciewicz RA, Singh D, Hussell T. Mucosal Immunol. 2015;8(5):1021-30.

Cell death: A double edged sword in lung repair

monoMDMAMΦ

CD36BAI1

TIM1TIM3

TIM4 AxlMerTK

Tyro3

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50

250

500

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Differential expression of TAM receptors on human macrophage subsets

Grabiec AM, Denny N, Doherty JA, ……Fowler SJ, Simpson A, Hussell T.Diminished airway macrophage expression of the Axl receptor tyrosine kinase is associated with defective efferocytosis in asthma.J Allergy Clin Immunol. 2017 Apr 12. (17)30585-7.

Cell death: A double edged sword in lung repair

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Apoptotic sensing by lung basal cells

Multitasking basal cells: combining stem cell and innate immune duties.Shaykhiev R. Eur Respir J. 2015 Oct;46(4):894-7

P63 Survivin DAPI

Sym

met

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mm

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P63 Survivin

8 days post influenza infection

Basal cells respond to local cues:

apoptotic cells

AXL drives basal cell proliferation

Rock JR, Randell SH, Hogan BL. Dis Model Mech. 2010 Sep-Oct;3(9-10):545-56. Airway basal stem cells: a perspective on their roles in epithelial homeostasis and remodelling.

Basal cell hyperplasia

Epithelial macrophage cross-talk via the Glial cell line-derived neurotrophic factor family

Key roles in the control of neuron survival and differentiation.

Distant members of the TGF-β superfamily

Neurturin influences inflammatory responses and airway remodeling in different mouse asthma models.Mauffray M, Domingues O, Hentges F, Zimmer J, Hanau D, Michel T. J Immunol. 2015;194(4):1423-33.

Neuturin and its receptor in the lung

Emma Connolly

Human Bronchial epithelial cells (Beas-2b)

Not all cells are affected equally

Dr Sylvia Liu

Most variable genes that define these clusters

Bind CXCR2Recruits Mono and neuts

IFN-response geneInduces apoptosisIn same group as IFITM3/M2

Where are the opportunities?

Health at different life stagesWhat drives change?

Infants

(n=20)

Adults

(n=20)

Age, median(range) 11 months (6-23) 59 years (36-78)

BAL fluid instilled (mL), median 15 (5-20) 50 (40-100)

BAL fluid retrieval (%), median 20 (5-40) 40 (20-66)

AM yield (cells x 105), median 2.9 (1.8-18) 2.35 (1.2-13)

Do the same pathways explain immune deficits in infants?

Matched for sex and ethnicityAll healthy diagnosis

Dr Anu GoenkaProf Peter Arkwright

Restoring tissue physiology to limit “inflammatory” disease

Inflamed tissues or trapped inflammatory cells?

Do our lungs become more stromal as we age

Is there a good and bad placental environment – training pre-birth.

Thanks to:Lab members: Dr Sylvia LiuDr Dave MorganDr Emma ConnollyDr Anu GoenkaDr Ben MulhearnDr Chris JaggerJakub ChudziakDr Christine ChewDr Oliver BlandMirium FranklinSarah Warbach

Past members:

Dr Alek GrabiecDr Naoya FujinoDr Toshifumi FujimoriDr Tom BellDr Manminder KaurDr Samira Salek-ArdakaniDr Mark FifeDr Lauren Cholewa

Dr Maryam HabibzayDr Alexandra Godlee Dr John Goulding Dr Robert SandersDr Robert Snelgrove

Clinical colleagues:Professor Angela SimpsonProfessor Stephen FowlerProfessor Rob NivenProfessor Jurgen Vestbo