Brigham & Women’s Hospital/Harvard Medical SchoolEdward NieuwenhuisArthur KaserStephanie DouganTakashi NagaishiMasaru YoshidaYvonne van de WalJonathan GlickmanSuzana BrozovicSean Colgan

CD1d: at the cusp of innate and adaptive immunity in regulating mucosal inflammationInflammatory Bowel Disease—Diagnostic and Therapeutic Strategies

Falk Symposium 154June 9, 2006

Beth Israel-Deaconess Mark Exley

Channing LaboratoryAndrew OnderdonkTetsuya Matsumoto

Gladstone InstituteStephen Young

NIHWarren StroberIvan Fuss

LIAIMitch Kronenberg

Kirin Breweries PharmaceuticalResearch Laboratories

SUNY DownstateMahmood Hussain

Induction of an Immune Response

Innate --

Immediate, hard-wired response between structural components of microbes and Pattern Recognition Receptorsof the immune system(e.g. peptidoglycan of bacteria⇔ NOD2 in monocytes andmacrophages → NFκB)

Adaptive (specific) --

Slower response obtained from interaction between an antigen presenting cell (DC,macrophage, B cell, IEC) and T cell that is associated with immunologic memory (e.g. nominal peptide of protein in association with MHC ⇔ T cell receptor on a T cell → Th1 or Th2 response)

Innate and AdaptiveImmune Responses Polarized Cytokine

Responses (Th1,Th2, Treg)

LeukocyteHoming/integrins(α4β7)

Tissue Destruction/Repair

CD1d, a MHC class I-related molecule on antigen presenting cells (APC) that presents glycolipids to natural killer T (NKT) cells for the regulation of

immune responses

αGalCer

αGalCer (α-galactosylceramide): a model glycolipid antigen

NKT cell

IFN-γIL-2

IL-4IL-5IL-13

Chemokines

Invariant TCR-α(Vα14-Jα18) chain

APCDC, Mφ, B cell,Hepatocyte,

IEC

CD1d

β2-microglobulin (β2m)

Th1

Th2

adapted from Nature Rev Immunology 2002;2:557

Intestinal Inflammation

T

Th2Th1

Microbiota/Dietary

Epithelium

Dendritic cells

Effector memoryT cell

Differentiated T cells

CytokinesIFN-γ ,TNF-α IL4,5,10,13

Microbial Colonization

IL-12 IL-4Differentiating factors—

Innate ImmuneSystem

Foreign(or host)glycolipids ?

Antigenpresenting

cell

CD1d

αGalCer

EBI3(IL-12-like)

NKT

IFN-γ, IL4,10,13

“Frontiers of the Immune System”

Heller et al Immunity, 2002; Nieuwenhuis et al PNAS, 2004

CD1d

UC

CD1d

ImmuneMediatedDiseases

AlveolarMΦ

MIP-2

CXCR2PMN

PseudomonasAirway Lumen NKT

IFN-γ

Immediate

Delayed

CD1d

DC

Nieuwenhuis E et al, Nature Med 2002

NKT

NKT

CD1 and CD1-restricted T cells mediate anendogenous pathway of antimicrobial defense

at the mucosal surface of the lung due to ability torecruit PMNs and activate macrophages

3 6 12 24 48 72

104

105

106

107

108

109

1010

CFU/g

<102

103

CD1d KO mice

Hours after innoculation

, P<0.05*

*

* * *

CD1d KO

Wild-type

Germ-free CD1d-deficient miceexhibit a major defect in early

colonization with E. coli §

§ E.coli strain ATCC 25922

105

106

107

108

1010

Jejunum Ileum Cecum Colon

PBSαGalCerCF

U/g

*: P<0.05109

104

103

102 ** **

**: P<0.01

**

αGalCer Treatment Prevents Mucosalcolonization of the intestine with E. coli

in germ-free wild-type mice

104

105

106

107

108

109

1010

Jejunum Ileum Cecum Colon

CD1 KO miceWild type mice

Viable E. coli in intestines of germ-freeCD1d-deficient mice and wild type micefour days after colonization suggests

a small intestinal defectCF

U/g

: P<0.05*

*

Altered ultrastructure of Paneth-granules in germ-free, CD1d-deficient mice

Wild Type CD1d-KO

Diminished degranulation of Paneth Cellsupon colonization with E. Coli in

germ free CD1d-/- mice

YXXZP

CD1d

IFN-γ IL-10X

YXXZ

IEL

YXXZ

NKT

Lumen

Serosa

Retrograde Signalling of CD1d on intestinalEpithelial Cells induces IL-10 secretion

P

Proc Natl Acad Sci USA 1999;96:13938; Gastroenterology 2003;124:1420;J Clin Invest 2004;113:1490; Nature Med 2004;10:535; J Exp Med 2005;202:529

MTP*

*MTP (microsomal triglyceridetransfer protein)

phospholipid

*24.7 NKT cell hybridoma; ND = not detectable

0102030405060708090

100

MODE-KMODE-KNKT

MODE-KNKT

αGalCer

IL-1

0 (p

g/m

l)

MODE-KNKT

αGalCeranti-CD1d

MODE-KNKT

αGalCerIgG2b

NKT cells* induce CD1d-restricted IL-10 secretion by MODE-K absorptive epithelial

cell line

020406080

100120140

MODE-K MODE-KNKT

MODE-KNKT

αGalCer

IL-1

0 (p

g/m

l) IL-10 siRNA

Control siRNA

ND

αGalCer

NKT cellMODE-K(IEC)

CD1d

anti-CD1d

SilenceIL-10

production

iNKT

Difference in retrograde CD1d signaling in dendritic cells and epithelial cells –

CD1d induces IL-12 from DC

0102030405060708090

100

Dendritic cell Dendritic cellNKT

Dendritic cellNKTαGC

IL-1

0 (p

g/m

l)

0100200300400500600700800

IL-1

2 (p

g/m

l)

Dendritic cell Dendritic cellNKT

Dendritic cellNKTαGC

ND ND ND

ND = not detectable

APC CD1d

TCR

barrier-protective/anti-microbial/anti-inflammatory

IL-10 (IEC)Defensins (Paneth Cells)

pro-inflammatoryIL-12 (from DC)

MTP

NKT cells regulate multiple different biologicalpathways in mucosal tissues by interactionswith discrete antigen presenting cell types

MTP

CD1dIL-10

Defensins

IL-12EBI3?

IL-13n = 45P<0.001

MTP KO

Wild-type

0

20

40

60

80

100

0 1 2 3 4days

Surv

ival

(%)

MTP

Therapeutic implications:•Target end-products• Differences in cell types

(ligands, theircellular quantities/qualities)