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