MUCOSAL IMMUNITY - Columbia University · mucosal immunity alessandra pernis p&s 9-435 x53763 [email protected] challenges faced by the mucosal system specialization of cells involved

MUCOSAL IMMUNITY

Alessandra PernisP&S 9-435

[email protected]

CHALLENGES FACED BY THE MUCOSALSYSTEM

SPECIALIZATION OF CELLS INVOLVEDIN MUCOSAL IMMUNITY

ORGANIZATION OF THE MUCOSALIMMUNE SYSTEM

CLINICAL IMPLICATIONS

DEFINITIONS

MALT= MUCOSA-ASSOCIATEDLYMPHOID TISSUE

MALT is the highly specialized immune systemwhich protects mucosal surfaces. The lymphoidelements associated with different mucosalsites share organizational as well as functionalsimilarities. It is the largest mammalianlymphoid organ system and in an adult itcomprises approximately 80% of alllymphocytes.

COMPONENTS OF THE MUCOSA-ASSOCIATED LYMPHOID TISSUE

Gastrointestinal tract (GALT) Bronchial Tree (BALT) Nasopharyngeal area (NALT) Mammary gland Salivary and lacrimal glands Genitourinary organs Inner ear

THE CHALLENGES

MOST FREQUENT PORTAL OF ENTRY FOR HARMFULSUBSTANCES. THUS THE MALT HAS TO MOUNT ANEFFECTIVE RESPONSE AGAINST A VAST NUMBER OFPOTENTIAL PATHOGENS.

THE MUCOSAL MEMBRANES OF THE DIGESTIVETRACT MUST ALLOW FOR THE ABSORPTION OFNUTRIENTS BY THE HOST. THUS THE MALT MUSTREMAIN HYPORESPONSIVE TO AN ENTIRE ARRAY OFHARMLESS SUBSTANCES.

SPECIALIZEDCOMPONENTS OF MALT

B CELLS

HUMORAL RESPONSES ARE CENTRAL TO ANEFFECTIVE MUCOSAL IMMUNITY.

THE MAIN HUMORAL MEDIATORS OFSPECIFIC MUCOSAL IMMUNITY ARESECRETORY IgA AND, TO A LESSER EXTENT,SECRETORY IgM.

THE NORMAL INTESTINAL MUCOSACONTAINS AT LEAST 20 TIMES MORE IgA+THAN IgG+ LYMPHOCYTES.

CRITICAL FEATURES OFSECRETORY IgA

RESISTANCE AGAINST COMMONINTESTINAL PROTEASES

INABILITY TO INTERACT WITHCOMPLEMENT OR CELLS IN A WAY TOCAUSE INFLAMMATION

MECHANISMS OF PROTECTION BYSIgA AT MUCOSAL SURFACES

INHIBITION OF ADHERENCE VIRUS NEUTRALIZATION NEUTRALIZATION OF ENZYMES AND

TOXINS IMMUNE EXCLUSION AND INHIBITION OF

ANTIGEN ABSORPTION

FACTORS CONTROLLING IgA ISOTYPESWITCHING

APC

CD4

B BIgA-J

B7

CD28 TCRMHC II

ACTIVATIONCYTOKINESECRETION

CD40LCD40

TGF-β

ACTIVATION ISOTYPE SWITCHING

PROLIFERATIONDIFFERENTIATION

IL-2/IL-4

IL-5

IL-6

IL-10

FACTORS CONTROLLING THESECRETION OF IgA: THE J CHAIN

THE J CHAIN IS A 15 KD POLYPEPTIDE THAT ISDISULFIDE-BONDED TO THE TAIL-PIECES OF BOTH IgMAND IgA

IgA SECRETING B CELLS IN THE BONE MARROW DO NOTEXPRESS THE J CHAIN AND THUS SECRETE IgAMONOMERS

THE MAJORITY OF IgA PRODUCING B CELLS IN THEMUCOSA EXPRESS THE J CHAIN AND THUS PRODUCEDIMERIC IgA

THE J CHAIN STABILIZES THE MULTIMERS AND ITAPPEARS TO DETERMINE THE POLYMERIC IgA AND IgMSTRUCTURE WHICH ALLOWS POLYMERIC Igs TO COMPLEXWITH THE SECRETORY COMPONENT

FACTORS CONTROLLING THESECRETION OF IgA: THE SECRETORY

PIECE (POLYMERIC Ig RECEPTOR)

IgA-JIgA-J

LAMINA PROPRIA MUCOSALEPITHELIAL CELL

LUMEN

SECRETED IgA

PROTEOLYTIC CLEAVAGE

DIMERIC IgA

SECRETORY COMPONENTWITH BOUND IgA

ENDOCYTOSED COMPLEXOF IgA AND SECRETORY COMPONENT

T CELLS

TH1 OR TH2?

MUCOSAL TISSUES % CD3+ TCR %CD4+ TH1:TH2 CD4:CD8αβ γδ

INDUCTIVE SITES PEYER’S PATCHES

EFFECTOR SITES LAMINA PROPRIA

INTRAEPITHELIUM

25-35

40-60 >95 1-5 60 1:2-3 2:1

80-90 35-45 45-65 5-10 1:1 1:7-8

>90 1-5 60 1:1 2:1

LAMINA PROPRIA LYMPHOCYTES

LYMPHOCYTES WHICH ARE SCATTEREDDIFFUSELY THROUGHOUT THE LAMINAPROPRIA OF THE INTESTINE. (LAMINAPROPRIA=LAYER OF CONNECTIVE TISSUEBETWEEN THE EPITHELIUM AND THEMUSCULARIS MUCOSA)

LARGEST SINGLE T-CELL SITE IN HUMANS.MOST OF THE T CELLS WITHIN THELAMINA PROPRIA ARE CD4+.

INTRAEPITHELIAL LYMPHOCYTES (IELs)

IELs ARE LYMPHOCYTES WHICH AREINTERSPERSED BETWEEN THE COLUMNAREPITHELIAL CELLS OF THE VILLI IN THESMALL AND LARGE INTESTINE

IN HUMANS, MOST OF THE IELs ARE CD8+T CELLS. APPROXIMATELY 10% OF IELs AREγδ CELLS

BOTH THE γδ AND THE αβ TCR+ IELs SHOWLIMITED DIVERSITY OF T CELL

IELs EXPRESS A NOVEL INTEGRIN TERMEDHML-1 (human mucosal antigen 1).

FUNCTIONAL PROPERTIES OF IELs.

FIRST IMMUNE CELL LINE OF DEFENSE INTHE INTESTINE

DISPLAY CYTOTOXIC ACTIVITY SECRETE LARGE AMOUNTS OF CYTOKINES

ESPECIALLY IFN-γ AND TNF-α MODULATE THE KINETICS OF EPITHELIAL

CELL RENEWAL PLAY A REGULATORY ROLE IN TOLERANCE

TO DIETARY ANTIGENS

ORAL TOLERANCE

ORAL TOLERANCE

ORAL ADMINISTRATION OF A PROTEINANTIGEN MAY LEAD TO SUPPRESSION OFSYSTEMIC HUMORAL AND CELL-MEDIATEDIMMUNE RESPONSES TO IMMUNIZATIONWITH THE SAME ANTIGEN.

POSSIBLE MECHANISMS:– INDUCTION OF ANERGY OF ANTIGEN-

SPECIFIC T CELLS– CLONAL DELETION OF ANTIGEN-SPECIFIC

T CELLS– SELECTIVE EXPANSION OF CELLS

PRODUCING IMMUNOSUPPRESSIVECYTOKINES (IL-4, IL-10, TGF-β)

REGULATORY T CELLS(CD4+)

TH3 CELLS: A POPULATION OF CD4+T CELLSTHAT PRODUCE TGF-β. ISOLATED FROM MICEFED LOW DOSE OF ANTIGEN FOR TOLERANCEINDUCTION

TR1 CELLS: A POPULATION OF CD4+T CELLSTHAT PRODUCE IL-10. CAN PRODUCESUPPRESSION OF EXPERIMENTAL COLITIS INMICE

CD4+CD25+ REGULATORY T CELLS: APOPULATION OF CD4+T CELLS THAT CANPREVENT AUTOREACTIVITY IN VIVO.

REGULATORY T CELLS

CD8+SUPPRESSOR T CELLS: THE FIRSTIDENTIFIED POPULATION OF REGULATORY TCELLS THOUGHT TO BE INVOLVED IN ORALTOLERANCE. THEIR FUNCTIONS ANDCHRACTERISTIC HAVE NOT BEEN CLEARLYDEFINED

γδ T CELLS: STUDIES IN MICE INDICATETHAT THEY HAVE AN IMPORTANT ROLE INSOME MODELS OF ORAL TOLERANCE.

EPITHELIAL CELLS

Scanning electron microscopy of a single microdissected dome (a) of a murinePeyer's patch. The M cells are identified by their relatively short, dark brushborder; they are restricted to the dome epithelium (upper half in b). Crypts(arrows) are opening to the cleft between the dome and the neighboring villi.

From: Gebert et al., Am. J. Pathol. 154:1573, 1999

M CELLS

CHARACTERISTICS OF M CELLS

M ("membrane-like") CELLS ARE SPECIALIZEDEPITHELIAL CELLS WHICH OVERLIE LYMPHOIDFOLLICLES DOMES ALONG THE LENGTH OF THE SMALLAND LARGE INTESTINE.

STRUCTURAL FEATURES INCLUDE:– FEW SHORT IRREGULAR MICROVILLI– ABUNDANT ENDOCYTIC VESICLES– LOW LYSOSOMAL CONTENT– DISTINCTIVE GLYCOCALIX– BINDING SITES FOR SECRETORY IgA BUT NO SC– POCKETS IN THE BASOLATERAL SURFACE

FUNCTIONS OF M CELLS

ANTIGEN SAMPLING

PORTAL OF ENTRY FOR SELECTEDPATHOGENS

Dendritic cell-mediated transport of commensalbacteria in the gut

From: Kraehenbuhl and Corbett, Science 303:1624, 2004

Dendritic cells can sampleAntigen indirectly via M-celltranscytosis (right) ordirectly via processes thatextend across the epithelialbarrier (left). DCs presentantigen to B- and T-cells,either directly within thelamina propia or followingtrafficking to the regionallymph nodes

Role of CX3CR1 in Luminal Sampling by Gut DCs

From: Niess et al. Science 307:254, 2005

CX3CR1 is a chemokine receptor whose ligand is an unusual chemokine; rather than secreted, it is membrane-bound

ORGANIZATION OFMALT

PEYER’S PATCHES

ORGANIZED MUCOSAL LYMPHOID FOLLICLESWHICH LACK AFFERENT LYMPHATICS.

PEYER’S PATCHES ARE FOUND IN THE SMALLINTESTINE.

FOLLICLES SIMILAR TO PEYER’S PATCHESARE FOUND IN THE APPENDIX, IN THEREST OF THE GASTROINTESTINAL TRACTAND IN THE RESPIRATORY TRACT.

From: Iwatsukit et al., Histochem. Cell Biol. 117:1363, 2001

Anatomy of a Peyer’s Patch

INDUCTIVE LYMPHOEPITHELIALTISSUES:

PEYER’S PATCHES

M CELLS

B

B

T

MESENTERIC LYMPH NODES

APC

ACTIVATEDLYMPHOID FOLLICLE

THORACIC DUCT PERIPHERALBLOOD

T

T

T

B

B

B

B

EFFECTOR SITES:LAMINA PROPRIA AND

INTRAEPITHELIUM

DISTANT GUTMUCOSA

PERIPHERAL BLOOD OTHER EXOCRINE TISSUES

T4

T4

T8T8

APC

IgA-JBSC SIgA

CLINICAL IMPLICATIONS

IgA DEFICIENCY

IT IS THE MOST COMMON PRIMARYIMMUNODEFICIENCY

IT IS USUALLY DEFINED BY A SERUM IgACONCENTRATION OF LESS THAN 50 µg/ml

IgA DEFICIENT INDIVIDUALS OFTEN APPEARPERFECTLY HEALTHY AND ARE IDENTIFIED UPON SERVING AS BLOOD DONORS UPON UNDERGOING ANAPHYLACTIC SHOCK WHEN

RECEIVING BLOOD TRANSFUSIONS

CLINICAL MANIFESTATIONS OFIgA DEFICIENCY

INCREASED INCIDENCE OF INFECTIONS UPPER AND LOWER RESPIRATORY TRACT GASTROINTESTINAL

HIGHER INCIDENCE OF AUTOIMMUNEDISEASES

HIGHER INCIDENCE OF ALLERGIC DISEASES

HIGHER INCIDENCE OF CELIAC DISEASE

INFLAMMATORY BOWEL DISEASE(IBD)

IBD IS A CHRONIC, RELAPSING AND REMITTINGINFLAMMATORY CONDITION

TWO OVERLAPPING PHENOTYPES: CROHN’S DISEASE (CD), WHICH AFFECTS THE DISTAL

SMALL INTESTINE AS WELL AS THE COLON IN ATRANSMURAL MANNER

ULCERATIVE COLITIS (UC), WHICH PREDOMINANTLYAFFECTS THE COLON IN A SUPERFICIAL MANNER

THE ETIOLOGY IS UNKNOWN: ? DUE TO A DYSREGULATEDMUCOSAL IMMUNE RESPONSE TO UNKNOWN ANTIGENSPRESENT IN THE NORMAL, INDIGENOUS BACTERIAL FLORA– MUTATIONS IN NOD2 (A CYTOSOLIC RECEPTOR FOR

PATHOGENIC BACTERIAL SIGNALS) INCREASE THE RISK OF CDBY A FACTOR OF 20-40.

TRAF6ECSIT

DD

TIR domain MyD88

IRAK

NF-κB

MEKK1

IKK-γ

p65

extracellular space

cytosol

TAK

IKK-α IKK-β

MD2

CD14

Adaptor proteins

Kinases

IKK complex

p50I-κB

Receptor Complex

Similarity Between TLR-4 and NOD2

TIR = Toll/IL-1 receptorDD = Death domainIKK = I-κB kinase

RICK

CARD

LigandRecognition

NOD2

TLR-4

IBD: IMMUNOLOGIC FEATURES

CELL-MEDIATED IMMUNITY (ACTIVE CD): INCREASED NUMBER OF ACTIVATED MUCOSAL T

CELLS SECRETING IFN-γ (TH1) INCREASED MUCOSAL PRODUCTION OF CYTOKINES

THAT ACTIVATE TH1 CELLS (IL-12 AND IL-18) DEFECTS IN REGULATORY (IL-10 PRODUCING) T

CELLS

IBD: IMMUNOLOGIC FEATURES

HUMORAL IMMUNITY: MASSIVE INCREASE INTHE NUMBER OF PLASMA CELLS AND IN IgGPRODUCTION (IgG2 IN CD AND IgG1 IN UC)

IMBALANCE OF PRO-INFLAMMATORY (TNF-α,IL-1,IL-8, IL-12) AND ANTI-INFLAMMATORYCYTOKINES (IL-10, IL-4, IL-13)

IBD:EMERGING BIOLOGICTHERAPIES

INHIBITORS OF PROINFLAMMATORY CYTOKINES– Anti-TNF therapies: infliximab

ANTIINFLAMMATORY CYTOKINES– IL-10– IL-11

ANTI-LEUKOCYTE ADHESION THERAPIES– Anti-α4 integrin: Natalizumab ?

INHIBITORS OF TH1 POLARIZATION– Anti-IL-12– Anti-IL-18– Anti-IFN-γ

CELIAC DISEASE

CELIAC DISEASE IS A T CELL MEDIATEDIMMUNE DISEASE OF THE SMALL INTESTINETRIGGERED BY GLUTEN

MAJOR FEATURES: VILLOUS ATROPHY WITH A LYMPHOCYTIC

INFILTRATE INCREASED EPITHELIAL PROLIFERATION

WITH CRYPT HYPERPLASIA MALABSORPTION

Normal Celiac pt

CELIAC DISEASE: IMMUNOLOGICFEATURES

ANTIGEN: GLUTEN (gliadin and glutenins)

IT IS ASSOCIATED WITH HLA-DQ2 OR HLA-DQ8RESTRICTED LAMINA PROPRIA CD4+ T CELLS THATRECOGNIZE GLUTEN AND SECRETE INTERFERON γ (98% OFPEOPLE WILL CARRY THESE HAPLOTYPES)

GLIADIN IS A SUBSTRATE OF TISSUE TRANSGLUTAMINASE(TRANSFORMS POSITIVELY CHARGED GLUTAMINES TONEGATIVELY CHARGED GLUTAMIC ACID)

INCREASED B CELL ACTIVITY– ANTIBODIES AGAINST GLIADIN (IgA-AGA, IgG-AGA)– ENDOMYSIAL ANTIBODY (IgA-EMA)– TISSUE TRASGLUTAMINASE (IgA-tTG)

The CD4 side of the story

CELIAC DISEASE: IMMUNOLOGICFEATURES

IMPORTANTLY THE HALLMARK OF CELIAC DISEASE ISINTRAEPITHELIAL INFILTRATION BY CD8+ T CELLS– DIFFERENT FROM IBD– IELs ARE BELIEVED TO PARTICIPATE IN THE

PATHOGENESIS OF CELIAC DISEASE BY MEDIATING THEDESTRUCTION OF THE EPITHELIUM

– RECENT EVIDENCE POINTS TO THE FOLLOWINGSCENARIO: GLUTEN ALTERS EPITHELIAL CELLS OF PATIENTS WITH

CELIAC DISEASE LEADING TO PRODUCTION OF IL-15 ANDTO THE EXPRESSION OF NON-CLASSICAL MHC CLASS IMOLECULES. IL-15 IN TURN LEADS TO THE EXPRESSION OFRECEPTORS ON IELS FOR THESE NON-CLASSICAL MHCMOLECULES AND TO THE ACTIVATION OF THE IELS, WHICHTHEN KILL THE EPITHELIAL CELL

The CD8 side of the story

MUCOSALIMMUNIZATION

MUCOSAL VACCINES

VACCINES AGAINST MUCOSAL INFECTIONS MUSTSTIMULATE THE MALT IN ORDER TO BEEFFICACIOUS

BECAUSE OF SUBCOMPARTMENTALIZATION WITHINTHE MALT, VACCINES MUST BE ADMINISTERED BYTHE APPROPRIATE ROUTE

NONREPLICATING ANTIGENS ARE OFTEN RELATIVELYINEFFICIENT IN YIELDING STRONG AND LONG-LASTING MUCOSAL ANTIBODY RESPONSES

MUCOSAL VACCINES

NEW STRATEGIES FOR ANTIGEN DELIVERY: LIVE ATTENUATED RECOMBINANT BACTERIA AND

VIRUSES WITH KNOWN MUCOSAL TROPISM PROTECTIVE VEHICLES, E.G. LIPOSOMES AND

BIODEGRADABLE MICROSPHERES MUCOSAL LECTIN-LIKE MOLECULES ENDOWED WITH

IMMUNOSTIMULATORY PROPERTIES, E.G. CHOLERATOXIN

MUCOSAL IMMUNOTHERAPY

STRATEGY TO ATTEMPT TO TREATILLNESSES RESULTING FROM IMMUNEREACTIONS AGAINST AUTOANTIGENSENCOUNTERED IN NONMUCOSAL TISSUES

HUMAN TRIALS HAVE BEEN CONDUCTED INMULTIPLE SCLEROSIS, RHEUMATOIDARTHRITIS, UVEORETINITIS, AND TYPE IDIABETES

MUCOSAL IMMUNOTHERAPY

POTENTIAL PROBLEMS: LIMITED SUCCESS IN SUPPRESSING THE

EXPRESSION OF AN ALREADY ESTABLISHEDIMMUNE RESPONSE

MASSIVE AMOUNTS OF TOLEROGENS AREREQUIRED

IMMUNOSUPPRESSIVE EFFECT IS OF SHORTDURATION