Master

course Advanced Immunology and Diseases December 3rd 2019 Lyon

Gut mucosal immunology

from physiology to Inflammatory Bowel Diseases

Pr Gilles BOSCHETTI
Lyon-Sud University Hospital
Gastroenterology Department
INSERM U1111 Lyon-Gerland
 Conflicts of interest
• Abbvie, MSD, Pfizer, Janssen, Tillotts, Sanofi, Takeda,
Ferring
90 % of the antigenes penetrate by
mucosal route… the rest (10%) after
skin break-in.

Surface :
- Skin = 2 m2
- Lung = 140 m2
- Intestine = 200 m2
 Mucosal Immunology = functionnal paradox
Virus Commensal bacteria
Pathogenic bacteria Food antigens
Parasites Environnemental antigens

MUCOSA

Physiological function : Tolerance:

Immunity Active phenomenon
External environment interface
Anti-infectious :
Food absorption
Gas exchange, etc… - innate
Permeability - acquired
 MALT = Mucosal Associated Lymphoid Tissue

GALT = gut-associated lymphoid tissue

BALT = bronchus-associated lymphoid tissue

NALT = nose-associated lymphoid tissue

LALT = larynx-associated lymphoid tissue

SALT = skin-associated lymphoid tissue

CALT = conjunctiva-associated lymphoid tissue

 Global organization of the MALT

Diffuse
lymphoid
tissue

Organized
lymphoid
tissue
 The intestinal mucosal immune system - GALT
Environmental Ags
- Dietary Ag / commensal bacteria
- Pathogens
- Vaccines

Homeostasis Danger signals (TLR-L, adjuvants):

• IL-10, • proinflamm. Cytokines/CC
• TGFß, • activation of Innate Immunity
• PGEs,
• retinoic acid … Breakdown of oral tolerance
Allergy
Oral Tolerance: Chronic inflammatory diseases
Maintains tissue integrity
Suppression of mucosal and systemic DTH responses
 What are Inflammatory Bowel Diseases ?
Chronic diseases affecting the gastro-intestinal tract, characterized
by recurring and chronic inflammation that normally evolves in a
waxing and waning pattern

2 TYPES OF DISEASE

CROHN´S DISEASE ULCERATIVE COLITIS

IBD – major differences between CD and UC
 Epidemiology – age at diagnosis
80
70
60
nb patients

50
40
30
20
10
0
10 20 30 40 50 60 70

Age at diagnosis
Whelan G. Gastroenterol Clin North Am 1990
 The Global Map of IBD in 2019
IBD is a Modern Disease of Modern Times that is Rooted in
the Industrialization of Society
>10 per100,000

<4 per 100,000 5-10 per 100,000

Cosnes et al. Gastroenterology 2010
Ng et al. Gut 2013
 Ulcerative Colitis in China
8000 7512

6000

No of
cases 4000
2506
2000

0
1981-1990 1991-2000
Jiang and Chui. World J Gastro 2001
 IBD as an intermittent disease

Inflammatory activity
(CDAI, CDEIS, CRP)
Disease Diagnosis Early
onset disease

Pariente B, et al. Inflamm Bowel Dis 2011

 Crohn’s disease as a progressive disease
Progression of digestive damage and inflammatory activity in a
theoretical patient with Crohn’s disease
Stricture

Inflammatory activity
Surgery

(CDAI, CDEIS, CRP)

Bowel damage

Fistula/abscess

Stricture

Disease Diagnosis Early

onset disease
Pariente B, et al. Inflamm Bowel Dis 2011
 New goal of treatment:
Blocking disease progression and damage

Stricture

Inflammatory activity
Surgery
Digestive damage

Fistula/abscess

Stricture

Disease Diagnosis Early

onset disease
Pariente B, et al. Inflamm Bowel Dis 2011
 New goal of treatment:
Blocking disease progression and damage

Inflammatory activity
Digestive damage

Disease Diagnosis Early

onset disease
Pariente B, et al. Inflamm Bowel Dis 2011
IBD = complex pathophysiology!!
IBD = complex pathophysiology!!
 Genetics
• Arguments in favor of genetic factors:
– Family forms
– Different prevalences by ethnic group
– Concordance in monozygotic twins (40%)
IBD = complex polygenic diseases
(~240 susceptibility loci)

from Lees, C.W., Gut 2011

Jostins et. al. Nature, 2012
IBD = complex polygenic diseases
IBD = complex pathophysiology!!
 Gut bacterial flora
Stomach 0-102
Lactobacillus Jejunum 102
Candida
Lactobacillus
Streptococcus Streptococcus
Helicobacter pylori

Duodenum 102 Proximal Ileum 103

Lactobacillus Lactobacillus
Streptococcus Streptococcus

Colon 1011-1012
Bacteroides
Distal Ileum 107-108 Clostridium groups IV&XIV
Streptococcus Bifidobacterium
Clostridium Enterobacteriaceae
Bacteroides
Actinomycinae
Corynebacteria
1014 bacteria (gut)
1013 cells (human body) modified from Sartor, 2008
 Birth : sterile digestive tract
Successive implantations of Environment
bacterial species
&
Host

Acquisition of an "adult" microflora (2-6 years)

Biodiversity ++
Stability ++
Fonctions = organe
 Gut bacterial flora = microbiota

Functions :
- Facilitates the digestion of non-assimilable foods for epithelial cells such as dietary
fiber

- Synthesizes many metabolites like vitamins (vitamins B and K)

- Degrades carcinogens such as nitrosamines

- Protects against pathogens invasion by occupying the intestinal lumen

- Important role in the development of infant immune responses

 Stability of the digestive microflora

In the same individual the flora is

stable over time ++ (" ID card ")

Destabilizing factors:
- Antibiotherapy
- Intestinal infection
- IBD flare-up

Resilience = return to basal state

Bacterial species abundance differentiates Distribution of non redundant bacterial genes
IBD patients and healthy individuals in IBD patients and healthy controls
Qin J et al., Nature 2010
Faecalibacterium prausnitzii : a major bacteria of the
normal gut microbiota belonging to the Firmicutes
Phylum
A lower proportion of F. prausnitzii on resected ileal
Crohn’s mucosa associated with higher risk of
endoscopic postoperative recurrence.
Sokol et al. PNAS 2008
Infectious pathways in Crohn's disease

Adherent Invasive E coli (AIEC)

Barnich et al. WJ Gastro 2007

Intestinal flora and IBD

Microbiota = therapeutic target

 Fecal transplantation
• 1st description in Chinese medicine 1700 years ago ...
• Principle: restoration of an "healthy" microbiota in the colon
• Administration of a stool suspension by naso-jejunal tube /
Rectal enema / Colonoscopy after colonic preparation
 In IBD encouraging results…
50

endoscopic remission (%)

Patients in clinical and
40
p = 0.03
30
24 %
20

10 5%
n = 9/38 n = 2/37
0

Fecal Placebo
Transplantation

o
T

eb
FM

ac
Pl
Significant increase in the rate of clinical and endoscopic remission at week 7 in the
fecal transplantation vs placebo group
Moayyedi P et al. Gastroenterology 2015
 In IBD encouraging results… but
Heterogeneous techniques and protocols
- High / low
- Endoscopy / enema / pills
- Duration

Long-term safety
- Microflora associated with diabetes, obesity, cancer…
- Infections
Simone et al. Science 2016
 In IBD encouraging results… but
Heterogeneous techniques and protocols
- High / low
- Endoscopy / enema / pills
- Duration

Long-term safety
- Microflora associated with diabetes, obesity, cancer…
- Infections

Selection of the donor (s)

- Costs
- "Good" donors
- Stool bank
 IBD = complex pathophysiology!!
Genetics Environment

Intestinal Intestinal flora

barrier
Epithelial
cells

Innate
Immunity Activation of
effector cells

Failure of immune
responses regulation
 IBD = complex pathophysiology!!
Genetics Environment

Intestinal Intestinal flora

barrier
Epithelial
cells

Innate
Immunity Activation of
effector cells

Failure of immune
responses regulation
The epithelial barrier
- First level of defense =
physicochemical barrier

- Rapid renewal of epithelial cells

(stem cells)

- Inter-cellular tight junctions

(claudine, occludin)

- Secretion of mucus and

formation of glycocalyx (goblet-
cells)

- Synthesis of anti-microbial
peptides (defensins produced by
Paneth cells)
 Abnormalities of the intestinal barrier

Toxic (DSS)
Colitis
N-cadherine -/-
Murine models

Alteration of the intestinal barrier =

massive entry of microorganisms then activation of GALT
(Gut Associated Lymphoid Tissue)
 Abnormalities of the intestinal barrier

Mucus Intestinal
Tight Apoptotic permeability
jonctions epithelial cells
n Role +++ of TNFα in alterations of the epithelial barrier Michielan et al. 2015
 Abnormalities of epithelial cells
Abnormality of
Endoplasmic Colitis
Reticulum stress
Murine models
Muc2

Inflammatory
mediators
Stress Activation NFkB
Accumulation UPR pathway
of proteins
Apoptosis
Endoplasmic reticulum
 Abnormalities of epithelial cells
• Paneth cells:
– Cells localized in crypts
– Secretory cells (lysozyme, cryptidines, defensins)
– Morphological and functional abnormalities during IBD
– Abnormalities of autophagy
 Abnormalities of epithelial cells
• Autophagy

Bel S et al. Sciences 2017

Immune response
- Second level of defense

- Small intestine = diffuse

lymphocytes, Peyer's patches,
draining lymph nodes
(mesenteric lymph nodes) and
isolated lymphoid follicles

- Colon = diffuse lymphocytes and

isolated lymphoid follicles

- Objective = to generate an
adaptive response against
pathogenic antigens
Cells of intestinal immunity

Neurath et al. Nat Rev Immunol 2014

Cells of intestinal immunity
Innate immunity cells

- Phagocytes = mainly
macrophages (derived from blood
monocytes). Some of them are
tolerogenic (production of IL-10)

- Innate Lymphoid Cells (ILC)

- ILC1 = NK
- ILC2 = production of IL-13, IL-
4 et IL-5
- ILC3 = RORgT, IL-23R and
production of IL-17 and IL-22

Neurath et al. Nat Rev Immunol 2014

Cells of intestinal immunity
Innate immunity cells

- Intra-Epithelial Lymphocytes = IEL

- TCR gd
- 1 IEL/10 epithelial cells
- Cytotoxic, anti-viral et anti-tumor
properties

- Dendritic cells
- Essential for adaptive immunity
- Several profiles : tolerogenic or
effector

Neurath et al. Nat Rev Immunol 2014

Cells of intestinal immunity

Neurath et al. Nat Rev Immunol 2014

Peyer’s patch = GALT inducing site
Peyer’s patch = GALT inducing site

M cells:
Without microvilli and mucus

Many microvesicles

Particular form: close contact with dendritic

cells, macrophages and lymphocytes at their
basement membrane.

Selectively capture microparticles, often

antigenic.

Ag cross their cytoplasm as vesicles

Peyer’s patch = GALT inducing site
Routes of mucosal antigen transport
 Oral tolerance
Goubier et al. Immunity 2008; Dubois et al. Gastroenterol. 2009
Ag Flora Essential role of :
- dendritic cells
MUCOSA - Foxp3+ regulatory T cells
- liver and MLN

Prevents priming of Ag specific

Free Ag
Ag loaded DC
Afferent Effector T cells and DTH responses
Blood lymphatics
drainage

pDC
CD4 +
Fox p3 -
MLN
CD8 DC
Gut
CD8 T cell CD4 +
Foxp3+ Treg
wall Fox p3 +
Deletion Conversion
pDC
Thoracic duct
CD8
CD4 +
LIVER CD8
Fox p3 +
CD4 +
Fox p3 +

Treg
SYSTEMIC LN suppression
 normal Crohn
Homeostasis Inflammatory cascade
Immune tolerance Breakdown of Immune tolerance
 Immune response and IBD
Mucosal inflammation

Innate immunity Adaptative immunity

abnormalities abnormalities
NOD and TLR receptors
Activation of signaling Excess effector T cells
pathways Failure of regulatory T cells
Adhesion molecules Oral tolerance abnormalities
Apoptosis
 NOD and TLR receptors
§ No spontaneous inflammation
§ Hyperplasia of GALT
§ Increased intestinal permeability
§ Excess of CD4 T cells producing TNFα and
NOD2 -/- mice IFNγ

§ NOD2 mutations
§ Decrease of TLR3
§ TLR4 increase
§ TLR9 inducing secretion of IL-8
IBD
Abnormal activation of signaling pathways

n Activation of NFkB and MAPK during IBD

¨ Production of pro-inflammatory cytokines
¨ T cells activation
n Efficacy in animals of signaling pathways inhibitors
n Deceptive effect in humans (functional duality ++)
Nenci A et al. Nature 2007
Increase of adhesion molecules

Lobaton et al. Aliment Pharmacol Ther 2014

Abnormalities of apoptosis during IBD

• Decreased apoptosis of T cells in lamina

propria

• CD = mitochondrial pathway
• UC = extrinsic pathway (Fas-FasL)

• Anti-TNF = T cells apoptosis induction

• Azathioprine = mitochondrial apoptosis
 Cytokines during IBD

Pro-inflammatory Anti-inflammatory
IL-1
IL-6 IL-1RA
IL-8 IL-10
TNFa TGFb
 Cytokines during IBD

TNFα, sources, target cells and mechanisms of action

Neurath et al. Nat Rev Immunol 2014
 Cytokines during IBD

« Th1 » murine models « Th2 » murine models

CD UC
Models Model
IL-10-/-, Scid, TNBS oxazolone
IFNg IL-4
IL-2 IL-13
 Cytokines during IBD

Acute lesions Chronic lesions

Th1 Th2

Th2 Th1
Different therapeutic strategies between
"recent" disease vs "old" disease?
 A little bit of « history… »
Th1
Tbet IFNγ
STAT4 IL-2
IL-12 _
+ IFNγ 1986
IFNγ
IL-4
Th2
_
IL-4
+ IL-4 GATA3 IL-5
STAT6
IL-13

Naive
CD4+ Tcells
 A little bit of « history… »
Th1
Tbet IFNγ
STAT4 IL-2
IL-12 _
+ IFNγ 1986
IFNγ
IL-4
Th2
_
IL-4
+ IL-4 GATA3 IL-5
STAT6
IL-13

Naive
CD4+ Tcells Th17
IL-6 IL-17
+ RORγt IL-21
TGFβ STAT3
IL-22
2005
+
IL-23
 Cytokines during IBD
Th17
IL-17
RORγt IL-21
IL-22
+
IL-23

IBD : Functions of Th17 cytokines:

ì Neutrophiles
ì LT Th17 ì Metalloproteases
ì Anti-microbial peptides
ì IL-17
ì Repair Molecules
ì IL-23 ì Defensins
ì RORgt
IL-23 and Th17 lineage

Boschetti G, et al. HGOD 2012

 IL-23 drives development of inflammatory pathogenic TH17 cells

Local TGF-β concentration

IL-23R
IL-23
iTreg
T cell activation
Antigen

APC Inducible TH17

+ TGF-β ↑ RORγt
TH 0
TH17 ↑ IL-23R
+ IL-6
↑ IL-17

IL-23 exposure needed for

(–) IL-23
development of inflammatory
TGF-β3 In absence of IL-23,
TH17 cells producing high levels of TH17 cells differentiate
IL-17, IL-22, IFN, and TNF into non-pathogenic
TH17 IL-17+ and IL-10+ cells
(+) IL-23

Pathogenic TH17 APC

Non-pathogenic TH17
↑↑ RORγt ↑↑ IL-17 RORγt+
↑↑ IL-23R ↑↑ IL-22 IL-17+
↑↑ GM-CSF ↑↑ IFNγ IL-10+
↑↑ TNF
Zuniga LA, et al. Immunol Rev. 2013;252:78–88; Gaffen SL, et al. Nature Rev Immunol 2014;14:585–600.
Adaptive immunity abnormalities

Neurath et al. Nat Rev Immunol 2014

Adaptive immunity abnormalities

Th1 Treg
Th2 Tr1
Th17 Effector cells Regulatory cells

Intestinal
Homeostasis

Inflammation Inflammation

Excess of Defect in
Effectors Regulators
 Adaptive immunity abnormalities
Th1
Tbet IFNγ
STAT4 IL-2
IL-12
+
IFNγ Th2
IL-4
+ IL-4 GATA3
STAT6 IL-5
IL-13
Naive T cells Th17
IL-17
+ IL-6 RORγt IL-21
STAT3
TGFβ IL-22
IL-26
Cytotoxic granules
IL-9
Effector T
Cells
CD4 or CD8 Cell death
Adaptive immunity abnormalities
1. DNBS-specific colitis
DNBS DNBS
sensitisation challenge 48hr
d0 d5

2. LCMV class I- peptide-specific colitis

P14 Tg mice GP-33
TcR Tg Va2 pulsed-BM-DC
LCMV GP33 class-I

48hr
d-1 d0 d7
Transfer GP33 GP33
TcRTg Va2 CD8T cells sensitisation challenge

3. ECOVA / OT-I x RAG°/°

OT-I X Rag2 -/- mice
TcR Tg Va2/Vb5 OVA/CFA
OVA peptid class-I

48hr Nancey S et al.

d-1 d0 d7 Gastroenterology 2006
Transfer OVA ECOVA* Nancey S et al.
TcR Tg Va2/Vb5 CD8+T cells sensitisation challenge
Inflamm Bowel Dis 2011
 Adaptive immunity abnormalities

Regulatory T cells

IL-2 -/-
TGF-β -/- Treg defect= Foxp3 mutation
IL-10 -/-
Murine models Scurfy gut inflammation
IPEX syndrom
Immune dysregulation, Polyendocrinopathy,
Enteropathy, X-linked
Colitis
Severe
enterocolitis

Fontenot et al. Nat Immunol 2003

Brunkow et al. Nat Genet 2001 Bennet et al. Curr Opin Pediatr 2001
 Adaptive immunity abnormalities
Regulatory T cells
Dendritic
Thymus cells Ag
Macrophage CD8+ T cells
Acide Rétinoic acid
IL-10
Naive CD4+ T cells
Thymocytes IL-2
TGFβ
CD4+ Foxp3-

Thymocytes
CD4+ Foxp3+

iTreg iTreg Tr1 iTreg Th3

Foxp3+ (IL-10 and TGFβ) (TGFβ)
« natural » Treg(nTreg)
« Induced » CD4+ Treg (iTreg) Treg CD8+ CD28low
CD4+ Foxp3+ CD25+
Shevach et al. Immunity 2006 Allez et al. Inflamm Bowel Dis 2004
 Adaptive immunity abnormalities
Induction of Treg (iTreg) in the gut
ü DC CD103+ in the gut and MLN
Dendritic cell
+++
MLN
DC CD103+ ü Conversion in presence of
iTreg TGFb and retinoic acid

avb8 ü TGFb needs an activation via

TGF-b
(avb8) integrin to be
Retinoic acid functionnal
Naive T cell
ü Defect in avb8 intégrin =
Coombes et al. J Exp Med 2007
colitis
Païdassi et al. Gastroenterology 2011
Sun et al., J Exp Med 2007 Worthington et al. Gastroenterology 2011
 Adaptive immunity abnormalities
CD4 Th1
Treg CD4 IFNγ
Foxp3+
CD4 Th17
IL-17
Regulatory cells CD8 cytotoxiques
Granzyme, Perforine
Effector cells

Dichotomy between Teff – Treg is more complex !

Detection of IL-17+ producing Foxp3+ Treg
Neoconversion of Treg in the gut
Voo et al. PNAS 2009
Sun et al. JEM 2007
 Adaptive immunity abnormalities

Lymphocyte plasticity
ü T cells can move from one
subpopulation to another

ü Description also of mixed

populations co-expressing
Foxp3 + IL-17 or Foxp3 + IFNg

ü Influence +++ of cytokines

present in the environment
(TGFb, IL-6…)
! Horwitz et al. Trends Immunol 2008
Mayne et al. Inf Bow Dis 2013 Zheng et al. J Immunol 2008
 Adaptive immunity abnormalities
Abnormality in number/proportion
Treg CD4
Foxp3+ Defect in suppressive function ?
Abnormalities of conversion in the gut
Neo-conversion in T effector cells
Treg
Therapeutic target ?
Cellular therapy
Medicines promoting the number /
function of Treg
 Conclusion
Environment

Intestinal Intestinal flora

barrier
Epithelial
cells

Innate
Immunity Activation of
effector cells

Failure of immune
responses regulation
 Anti-TNF Antibodies in IBD

Chimeric Human Humanized Human

monoclonal recombinant Fab’ recombinant
antibody antibody fragment antibody

VL VH

CH1
No Fc

PEG

IgG1 IgG1 IgG1

PEG

infliximab adalimumab certolizumab golimumab

pegol (UC only)
(CD only)
Mouse
Human
The problem with anti-TNFS: Infliximab
discontinuation due to treatment failure

Turner D, Lev-Tzion R. Dig Dis Sci 2013;58:604–607

 Immunotherapy of IBD : many years of frustration !

Danese S. Gut. 2012;61(6):918-932.

 Reason 1: rely too much on
animal data

Animal models of immunopathogenesis

continue to have
significant limitations in terms
of human translation
C3H/HeJBir mice
 Primary endpoint analysis at week 6 shows no
treatment benefit of Secukinumab (anti-IL17A)

secukinumab
placebo

Hueber W al. Gut 2013

Reason 2: extrapolate from
other diseases
 Abatacept in methotrexate-resistant active RA
35
Abatacept (N = 424)
30 **
Placebo (N = 214)

25 ** **
Responders (%)
**
20
**
15
** *
10

0
15 1 29 57 85 113 141 169 197 225 253 281 309 337 365
Visit (days)

Kremer JM et al. Ann Intern Med 2006

 Abatacept for active Crohn’s disease

30
Patients in remission
25

20
(%)

15

10 6.3%
5
4.7% 3.9%
0
Placebo 3mg/kg 10mg/kg
abatacept abatacept

Sandborn WJ et al. NEJM 2012

Since 2015 : years of success for IBD!

Tofacitinib
Ustekinumab
Vedolizumab
Leukocyte Trafficking as a Target in Inflammatory
Bowel Disease

Rutgeerts P. Gastroenterology 2009;136:1182–1197

Therapeutic Targets for
Lymphocyte Trafficking

Lobaton T et al AP&T 2014; 39: 579-594

93
 S1P1 Receptor Modulator (Ozanimod) Mechanism
of Action
Agonists of S1P1 receptor:
•Binding induces receptor
internalization – leads to “functional
antagonism”
Downstream effects:
•Transiently reduces lymphocyte
trafficking
•Suppresses inflammatory cytokines
•Stabilizes epithelial barrier function
Ozanimod: Investigational S1P1
receptor modulator
•Binds to the S1P1 receptors on naïve
Adapted from Horga A and Montalban X. Expert Rev Neurother. 8(5): 699-714;
and central memory T and B
008. Degagne E, Saba JD. Clinical and experimental gastroenterology.
2014;7:205-214. Gonzalez-Cabrera PJ, et al. F1000 prime reports. 2014;6:109.
Since 2015 : years of success for IBD!

Tofacitinib
Ustekinumab
Vedolizumab
 Ustekinumab Background
• Ustekinumab is a fully human
IL-12 IL-23
IgG1k monoclonal antibody
p40
p35 p19 p40
that binds the p40 subunit of
ustekinumab
Interleukins-12 & 23
• Prevents IL-12 and IL-23 from
binding IL-12Rb1
NK or T cell
membrane
• Normalizes IL-12 and IL-23
No IL-12 or IL-23 mediated signaling, cellular
Intracellular signal activation, and cytokine
production
• Approved for psoriasis and
Since 2015 : years of success for IBD!

Tofacitinib
Ustekinumab
Vedolizumab
 Binding of cytokine receptors by cytokines
activates JAK pathways signaling
Cytokine binding to its cell surface
receptor leads to receptor
polymerization and activation of
associated JAKs

Activated JAKs phosphorylate the

receptors that dock STATs

Activated JAKs phosphorylate

STATs which dimerize and move to
the nucleus to activate new gene
transcription
JAK=Janus Kinase, P=Phosphate,
From Shuai K, Liu B. Nat Rev Immunol. 2003;3:900-11.
 Phase 2 Study of Tofacitinib (CP-690,550), an
Oral JAK 1/3 Inhibitor, in Active Ulcerative Colitis

99
Sandborn WJ, et al. N Engl J Med 2012;367:616-24
2019...: what’s next ?
 Can one determine a prognosis to help identify the best
treatment strategy for the individual patient?
Assessing prognosis at an early stage is essential for the development of an
appropriate management plan

Indolent Aggressive
Avoid intensive therapy, Assure early intensive therapy
immunosuppression, to avoid complications
adverse events
102
Martin JC . et al. Cell 2019 : 178;1473-1508
 Context and study objectives

• Response to different biotherapies in Crohn's

disease despite identical phenotypes

→ Likely different inflammation pathways

• Need for predictive response biomarker

• Single Cell RNA Seq +++

Hwang B. et al. Experimental & Molecular Medicine 2018

Patients & Methods
• 1st Step:
– 11 pieces of ileocecal resection of Crohn's
disease: ileal (93%), stenosing (86%), under
anti TNF at the time of surgery (71%)
– Cellular tissue analysis (lamina propria) in
adjacent inflammatory and non-inflammatory
zone and blood by scRNA seq and CyTOF
– Identification of% of cell types and cytokine
pathways
• 2nd step :
– Validation on 4 independent cohorts of
patients with Crohn's disease (441 patients)

Martin JC . et al. Cell 2019 : 178;1473-1508

 Results
Correlation between presence of GIMATS and response
Proportions of the different cells (tissue, blood)
to TNF antagonists

1. Discovery of GIMATS:

• cell organization with specific cytokine network

• Plasma cells, T cells, stromal cells, blood monocytes

• not correlated with blood biomarkers (CRP, VS)

2. Association of this GIMATS cell group with anti TNF alpha resistance
GIMATS profile Martin JC . et al. Cell 2019 : 178;1473-1508
Conclusion

• Identification of a specifically functioning cell

network called GIMATS in patients with
Crohn's disease resistant to anti-TNF

• Considerable therapeutic potential:

→ new biomarker with response to TNF

antagonists

→ Identification of new therapeutic pathways

Martin JC . et al. Cell 2019 : 178;1473-1508

 Anomalies de la tolérance orale
Ag

Ag

normal Crohn
Flore hétérologue
Flore autologue
LT
Luminal Flore autologue Flore hétérologue
LT LT LT
flora
réactif tolerance
aréactif réactif réactif
breakdown
Duchmann et al. Clin exp Immunol. 1995
L’immunité humorale
- Les lymphocytes B se
différentient en plasmocytes
sécrétant des IgA sous l'influence
de différentes cytokines,
notamment IL-6.

- Les IgA dimériques sont ensuite

transportées à travers l'épithélium
par le récepteur polymérique des
immunoglobulines (pIgR) et
libérées sous forme d'IgA
sécrétoires (S-IgA) dans la lumière

- Possibilité de trancytose inverse

ILC3 = Innate Lymphoid Cells 3

Zeng et al. Cell Death and Disease . 2019.

 ILC3 = Innate Lymphoid Cells 3

• Group 3 innate lymphoid cells (ILC3) are responsible for gastrointestinal mucosal homeostasis through
moderate generation of IL-22, IL-17 and GM-CSF in the physiological state.
• ILC3 contribute to the progression and aggravation of IBD by the dysregulation of NCR− ILC3 or NCR+ ILC3
function and the bias of NCR+ ILC3 towards ILC1 under the stimulation of IL-12 generated by CD14+ dendritic
cells as well as regulatory ILC dysfunction in the pathological state.
• The dysregulation of ILC3 results in overexpressions of inflammatory cytokines IL-22, IL-17 and IFN-γ, in which
IL-17 can recruit neutrophil cells to disrupt E- cadherin and junctional adhesion molecule-like molecule (JAML),
leading to the enhancement of epithelial permeability. Zeng et al. Cell Death and Disease . 2019.
ILC = Innate Lymphoid Cells

Zeng et al. Cell Death and Disease . 2019.