Ludvig M. Sollid Director (CIR), Professor (UiO)

Rationales behind novel therapies for coeliac disease

Coeliac UK Research Conference London, March 11, 2015

CONFLICT OF INTEREST

Ludvig M. Sollid

Ø  Regeneron Pharma – Consulting / Funding Ø  ImmusanT Inc – Membership on advisory committee / Honoraria Ø  Alvine Pharma – Membership on advisory committee / Honoraria Ø  Glenmark Pharma – Consulting

Magliocca et al Arch Histol Cytol 1992

l + gluten - gluten

NORMAL MUCOSA CELIAC DISEASE MUCOSA

+ gluten - gluten

NORMAL MUCOSA CELIAC DISEASE MUCOSA

antibodies to gluten (gliadin)

transglutaminase 2 (TG2)

Photo: Per Brandtzaeg

Koblingsanalyse

GENOME WIDE ASSOCIATION STUDIES

Mathew, Nat Rev Genet 2007

S

GWAS and MHC (HLA)

Slide courtesy: TH Karlsen

HLA association in celiac disease

DQ2.5  

DQ2.2  

DQ8  

60-90%

5-30%

1-5%

1-5%

TCR MHC II

T APC

T-cell Antigen Presenting Cell

Antigen

Ag

B

B-cell

Plasma cell

BCR

Antibodies

APC

IFN-γ IL-21

Wheat

TG2

Deamidation by TG2

The celiac lesion

CD4

CD4

CD4

APC

CD8 CD8

TCR

CD4 HLA-DQ2 (or -DQ8)

CD4 T cell

APC

CD8 T cell

Enterocyte

NKG2D

MIC

TCR cyto-lysis

Hüe et al Meresse et al

Immunity 2004

IL-15

   

Gluten  pep/de  

HLA-­‐DQ2.5  

Stanford  &  Oslo  

A gluten peptide caught in the act

α2-gliadin (AJ133612) 1 MVRVPVPQLQ PQNPSQQQPQ EQVPLVQQQQ FPGQQQPFPP QQPYPQPQPF PSQQPYLQLQ 61 PFPQPQLPYP QPQLPYPQPQ LPYPQPQPFR PQQPYPQSQP QYSQPQQPIS QQQQQQQQQQ 121 QQKQQQQQQQ QILQQILQQQ LIPCRDVVLQ QHSIAYGSSQ VLQQSTYQLV QQLCCQQLWQ 181 IPEQSRCQAI HNVVHAIILH QQQQQQQQQQ QQPLSQVSFQ QPQQQYPSGQ GSFQPSQQNP 241 QAQGSVQPQQ LPQFEEIRNL ALETLPAMCN VYIPPYCTIA PVGIFGTNYR

LQLQ 61 PFPQPQLPYP QPQLPYPQPQ LPYPQPQPF

digestive enzymes Transglutaminase (QXP)

LQLQ

61 PFPQPELPYP QPELPYPQPE LPYPQPQPF

after transglutaminase treatment

peptide (33 amino acids)

Generation of T-cell epitopes in the gut

PFPQPELPY PQPELPYP Q

PYP QPELPY P QPELPYPQ

PYPQPE LPY PQPE LPYPQ

6 copies of T cell epitopes

Shan et al, Science 2002; Arentz-Hansen et al, Gastroenterology 2002

DQ2.5

DQ8

HLA-DQ2.5 and HLA-DQ8 restricted gluten T-cell epitopes

T cell

S-PE B

B

B

B

HLA-DQ2.5-gluten epitope tetramers

Increased frequency of gluten-specific effector-memory CD4+ T cells in celiac disease

Control

TCD

UCD EM

Christophersen et al., UEG J 2014

T cells (CD3) Plasma cells (CD138)

Photo: Ann-Christin Beitnes

Active celiac disease

bio$n-­‐TG2          an$-­‐CD138          merge  

Visualization of TG2-specific plasma cells by immunofluorescence on

intestinal cryosections celiac

Di Niro et al., Nat Med 2012

Detection of TG2-specific plasma cells by flow cytometry on intestinal cell suspensions

biot

in-T

G2

Anti-IgA (gated on CD138+ plasma cells)

3.5 8.3 0.1 0.3 0.4 0.7

Patient with active CD Healthy donor

CD patient on GFD

Di Niro et al., Nat Med 2012

SA PE

PE b TG2

b TG2 b TG2

b TG2

APC

IFN-γ IL-21

Wheat

TG2

Deamidation by TG2

CD4

CD4

APC

CD8 CD8

B CD4

CD4

APC B CD4

APC

The celiac lesion

Mucosa (effector site)

Mesenteric lymph node or Peyer’s patch (priming site)

CD4

via blood via lymph

plasmablast

CD4 Germinal centre

CD8

IL-15

TCR MHCII

Ag

Relationship T-cell epitope and B-cell epitope

T cell help

Accessory molecules

Cytokines

T CD4 B

T-cell epitope must be physically linked with B-cell epitope prior to Ag processing

T-cell help to TG2-specific B cells by gluten-specific T cells

Gluten peptide

TG2 TG2-gluten peptide

complex

‘Hapten-carrier like’ model

TG2-specific B cell

TCR

MHCII

B

T-cell help

APC

Gluten-specific T cell

T CD4

HLA-DQ2

Anti-TG2 antibodies

Activation Proliferation Plasma cell differentiation

Activation Proliferation Clonal expansion

T-cell – B-cell interaction: An amplifying loop for the T-cell reponse

Gluten peptide

TCR

MHCII

B T

CD4

TG2

TG2

Gluten peptide

TCR

MHCII

B T

CD4

TG2

TCR

MHCII

B T

CD4

Deamidated gluten peptide

Transglutaminase (TG2) specific B cell

Deamidated gluten specific B cell

Gluten specific T cells can give help to both TG2-specific and deamidated gluten-specific B cells

Anti-TG2 antibodies

TG2

Anti-deamidated gluten antibodies

Key event in the pathogenesis of coeliac disease

Clonal expansion of:

- Gluten specific CD4+ T cells

- TG2 specific B cells

- Gluten specific B cells

The point of T- and B-cell clonal expansions is when the patient is crossing the bridge.

Non-dietary therapies for coeliac disease I

Prevent gluten from stimulating T cells •  Grain modification

•  Polymers binding gluten (BioLine Rx)

•  Glutenases (Alvine)

•  Epithelial barrier (Alba) •  Transglutaminase (TG2) inhibitors (Dr. Falk, Sitari)

•  Elafin (prevents gluten deamidation)

Non-dietary therapies for coeliac disease II

Modify immune response •  Cytokine therapy (anti-IL15; Celimmune, Calypso)

•  Peptide therapy (ImmusanT)

CONTRIBUTORS Oslo Roberto Di Niro Øyvind Steinsbø Rasmus Iversen

Luka Mesin Shuo-Wang Qiao

Asbjørn Christophersen Michael Bodd Elin Bergseng

Siri Dørum Jorunn Stamnaes Knut E.A. Lundin Ludvig M. Sollid

Chicago Carole Dunand Patric C. Wilson

Stanford Chaitan Khosla

ACKNOWLEDGEMENTS