Anne Müller Institute of Molecular Cancer Research

University of Zürich, Switzerland

Host and bacterial determinants of H. pylori persistence and gastric

pathogenesis

Helicobacter pylori persistence and immunomodulation

Anne MüllerInstitute of Molecular Cancer Research

University of Zürich, Switzerland

Helicobacter pylori resides in the human stomach, where it can cause gastritis, peptic ulcer disease and gastric cancer

Carriers with clinically overt disease (10‐20% of infected population)

H. pylori on gastric epithelial cells

Helicobacter pylori resides in the human stomach, where it can cause gastritis, peptic ulcer disease and gastric cancer… or remain asymptomatic

Carriers with clinically overt disease (10‐20% of infected population)

Asymptomatic carriers (>80% of infected population)

Helicobacter pylori resides in the human stomach, where it can cause gastritis and peptic ulcer disease… or remain asymptomatic

Carriers with clinically overt disease (10‐20% of infected population)

modeled by adult infection

Asymptomatic carriers (>80% of infected population)

modeled by neonatal infection of mice 

in humans: Harris et al., Gastroenterology 2008Robinson et al., Gut 2008

in mice: Arnold et al., Gastroenterology 2011

1 month post infection

Oertli et al. PNAS 2013

The H. pylori virulence/persistence factors VacA and GGT are required for DC tolerization, regulatory T-cell differentiation and persistence

VacA promotes the generation of peripherally inducedpTregs in the gastric lamina propria and their enrichment in the lung

0

5

10

15

% o

f CD4

T C

ells

pTregs

**

ns**

Uninf. WT VacAH.p. iN

0

2

4

6

8

10

% o

f CD4

T C

ells

RORgt+ pTregs

*

ns

*

Uninf. WT VacAH.p. iN

pTregs: CD4+ Foxp3+ neuropilin- RORt+ Tbet+/-

tTregs: CD4+ Foxp3+ neuropilin+

stomach

lung

0

20

40

60

80%

of C

D4 T

Cel

ls

Tregs

**

nsns

Uninf. WT VacAH.p. iN

Altobelli et al. MBio 2019

gastric Tregs

pulmonary Tregs

VacA promotes the generation of peripherally inducedpTregs in the gastric lamina propria and their enrichment in the lung

0

5

10

15

% o

f CD4

T C

ells

pTregs

**

ns**

Uninf. WT VacAH.p. iN

0

2

4

6

8

10

% o

f CD4

T C

ells

RORgt+ pTregs

*

ns

*

Uninf. WT VacAH.p. iN

pTregs: CD4+ Foxp3+ neuropilin- RORt+ Tbet+/-

tTregs: CD4+ Foxp3+ neuropilin+

stomach

0

20

40

60

80%

of C

D4 T

Cel

ls

Tregs

**

nsns

Uninf. WT VacAH.p. iN

gastric Tregs

VacA interacts with various myeloid cells in the gastric LP and promotes a tolerogenic transcriptional program

gastric lamina propria Peyer’s patches

0

1

2

3

4

Gen

e ex

pres

sion

(rela

tive

to H

PRT)

IL-10

PBS VacA VacA6-27

** **

ns

0.0

0.1

0.2

0.3

0.4

IL-12A

PBS VacA VacA6-27

ns nsns

0.0

0.2

0.4

0.6

0.8

PTGS2

PBS VacA VacA6-27

* *ns

0.0

0.5

1.0

1.5

2.0

TLR2

PBS VacA VacA6-27

ns **

qRT-PCR of CX3CR1+ macrophagesafter VacA exposure

qRT-PCR of CX3CR1+ macrophagesafter live Hp exposure

Altobelli et al. MBio 2019

VacA interacts with various myeloid cells in the gastric LP and promotes a tolerogenic transcriptional program

gastric lamina propria Peyer’s patches

qRT-PCR of CX3CR1+ macrophagesafter VacA exposure

qRT-PCR of CX3CR1+ macrophagesafter live Hp exposure

0

1

2

3

4

Gen

e ex

pres

sion

(rela

tive

to H

PRT)

IL-10

PBS VacA VacA6-27

** **

ns

0.0

0.1

0.2

0.3

0.4

IL-12A

PBS VacA VacA6-27

ns nsns

0.0

0.2

0.4

0.6

0.8

PTGS2

PBS VacA VacA6-27

* *ns

0.0

0.5

1.0

1.5

2.0

TLR2

PBS VacA VacA6-27

ns **

MacrophagesCD11b+

DCs

CD103+

CD11b-

DCsMHCII+

monocytes

PMSS1 RFP+

PMSS1 RFP-

CD11c

RFP

CD11b+ DCs and monocytes and macrophages, but not CD103+ DCs, encounter H. pylori in the infected gastric mucosa

02468

10 + H. pylori RFP

Uninf

Macro-phages

CD11b+

DCsMHCII+mono

CD103+

CD11b+

DCs

CD103+

CD11b-

DCs

+ H. pylori WT****

*****

*****

*

% R

FP+

cells

Arnold et al. Cell Reports, 2017

H. pylori is associated with several gastric disordersH. pylori is inversely associated with allergies and chronic inflammatory diseases

Hp+

Hp-

allergic asthmahay feveratopic dermatitisIBDceliac disease

Chen & Blaser, 2007; Reibman et al., 2008; Chen & Blaser, 2008; Amberbir et al., 2011, 2014, Genta et al, 2015

Neonatally infected mice are protected against clinical parameters ofovalbumin-induced asthma/allergic airway disease

Arnold et al. J. Clin. Invest. 2011

0 14 28-30Day

OVA/PBS+Alum i.p.

32

uninf iN / iA

1%OVAaerosol

sacrifice

sensitization challenge+ = allergic asthma

Neonatally infected mice, but not mice infected as adults, have elevated frequencies of pTregs in their lungs

VacA is required for infection-induced asthma protection and induction of allergy-suppressing Tregs

VacA confers asthma protection when administered in purified form, beginning in early life

Helicobacter pylori colonization confers protectionagainst allergic asthma (and IBD) through its immunomodulator VacA

Arnold et al., JCI 2011, Oertli et al., JCI 2012, PNAS 2013 Engler et al., PNAS 2014, Infl. Bowel Diseases 2015Meta‐analyses: Castaño‐Rodríguez et al. Gut 2017, Chen et al. Ann Allergy Asthma Immunol. 2017

Trans-maternal Helicobacter pylori exposure in utero and/or during lactationhas protective effects on offspring

Andreas Kyburz

Kyburz et al. JACI 2018

Helicobacter pylori VacA has protective effects on offspring exposed during lactation

Kyburz et al. JACI 2018

Sensitization Challenge EndpointBirth

Lactation

Weaning

VacA treatment of mother

House dust mite (HDM)-induced asthma model

Antibiotic exposure during the second week of life has the opposite effect on pTreg frequencies (and allergic asthma)

Control Ampicillin0

20

40

60

80

100

% F

oxP3

+ T c

ell

Nrp-

*

Control Ampicillin0

20

40

60

% F

oxP3

+ T c

ell

Nrp- RORt+*

Control Ampicillin0

20

40

60

80

100

% F

oxP3

+ T c

ell

Nrp- Tbet+**

Colonic Tregs:

with T. Borbet and M. Blaser

In the absence of BATF3-dependent CD103+ DCs, there is less Th1 recruitment to infected tissues => infection control is impaired

0

20

40

IFN-+

WT Batf3-/-

+Hp 4mthsWT Batf3-/-

+Hp 1mth

** *

% o

f TCR

/

+

CD

4+T

cells

103

104

105

106

107

Colonization

WT Batf3-/-

+Hp 4mthsWT Batf3-/-

+Hp 1mth

** **C

FU/s

tom

ach

Gate: Live CD45+ CD11c+

MHCII+ F4/80-

WT Batf3-/-

9.2

CD11b

CD

103

21143.6

0.00.20.40.60.8

CD103+ CD11b- DCs** *

WT Batf3-/-

+Hp 4mthsWT Batf3-/-

+Hp 1mthWT Batf3-/-ce

lls x

104

0123

Cxcl9

WT Batf3-/-

+Hp 4mths

*

Gen

e ex

pres

sion

(rel

ativ

e to

Hpr

t)

01234

Cxcl10

WT Batf3-/-

+Hp 4mths

**

010406080

100

% o

f Tre

gs

Nrp-1- pTreg*******

+Hp 1mthWT Batf3-/- WT Batf3-/-

*

0

20

40

60

% o

f Tre

gs

Tbet+ pTreg ***

+Hp 1mthWT Batf3-/- WT Batf3-/-

BATF3-dependent DCs are required for pTreg recruitment to the H. pylori-infected gastric mucosa

0

10

20

30

% o

f Tre

gs

Tbet+Nrp-1- pTreg ***

+Hp iNWT Batf3-/-

+Hp iAWT Batf3-/- WT Batf3-/-

*

lungstomach

WT Batf3-/-

Gate: Live CD4+ FoxP3+Tbet

Neu

ropi

lin-1

12 13

Uninfected

Gate: Live CD4+

FoxP3+Tbet

Neu

ropi

lin-1

26 5

WT Batf3-/-+Hp iN

BATF3-dependent DCs promote effector and regulatory T-cell recruitment to infected tissues through two distinct mechanisms

‐VacA is a persistence determinant that interacts with myeloid cells in the gastric LP and skews T‐cellresponses towards Tregs

‐direct and trans‐maternal exposure to H. pylori (or VacA) has trans‐generational protective effects inmodels of allergen‐induced asthma

‐H. pylori induces pTregs that express Tbet and RORt and follow a chemokine gradient to infected tissues

‐CD103+ DCs have a non‐redundant role in T‐cell recruitment (effector T‐cells and Tregs, to infected andtumor tissues and distant sites)

‐H. pylori interacts with eosinophils in the gastric lamina propria

Conclusions

Chronic Helicobacter-induced inflammation results inpreneoplastic gastric pathology

Epithelial hyperplasia

Intestinal metaplasia

Toller et al., PNAS 2011

H. pylori induces DNA double strand breaks in the nuclear genome of its host cells

H.p.

ctrl.

53BP1 H2AX merge with DAPI

Screening for H. pylori factors involved in DNA double strand break induction reveals a role for the type IV secretion system (T4SS)

Hartung et al., Cell Reports 2015

H. pylori-induced DNA DSBs require transcription, a functional type IV secretion system and NF-kB signaling

Hartung et al., Cell Reports 2015

AcknowledgementsFormer lab members:Ayca SayiIsabella Toller Vanessa CraigIris HitzlerEsther KohlerMathias OertliLivia GrüterManuel NobenDaniela EnglerMara HartungKatrin KochAndreas Kyburz

Current lab members:Michael BauerMariela ArtolaHind HashwahIsabelle Arnold Karelia VelezAngela FalleggerAleksandra AltobelliKatrin BertramAnna StellingXiaozhou ZhangAlessandra Gurtner

Key Collaborators:

Pavel Jansczak, IMCRNina Salama, FHCRCThomas Meyer, MPIIBSina Bartfeld, Univ. WürzburgTimothy Cover, Vanderbilt UnivMartin Blaser, NYU