Digestive, nutrition and metabolism
Immune System
DR.PETER K.CHEBOSS.
Department of immunology
Moi university school of medicine
Mucosal Immunology
- Lecture Outline -
I. Introduction.
II. Mucosa-associated lymphoid tissue (MALT)
III. Induction of mucosal immune responses.
IV. Lymphocyte trafficking and common mucosal
immunity.
V. Unique features of IgA immunity
VI. Mucosal T cells.
VII. Oral Tolerance.
VIII. Conclusion
Mucosal surfaces such as the gut are heavily
challenged by pathogens. The challenge to host
defense: protect against and clear infection; do not
respond to harmless antigens (food); effect host
defense without damaging the mucosal surface.
Non-antigen specific mechanisms are important but
sometimes insufficient for mucosal host defense.
Mucosal Immunology - Introduction
• Mucosal immunity protects internal
epithelial surfaces.
• Components of the mucosal immune
system include lymphoid elements
associated with internal surfaces of the
body (GI, respiratory, urogenital) and
exocrine secretory glands linked to these
organs, such as the salivary, lachrymal,
pancreas, and mammary glands.
Mucosa-associated lymphoid tissue
(MALT)
Examples:
- Nasal-associated lymphoid tissue (NALT).
- tonsils, adenoids.
- Gut-associated lymphoid tissue (GALT).
- Peyer’s patches.
- Bronchus-associated lymphoid tissue (BALT)
Mucosal Surfaces
The GI mucosal surfaces cover 400 m²
Thin – facilitate nutrient absorption.
The Gut Associated Lymphoid Tissue (GALT)
- Organized T and B cell areas
- where antigen is collected and adaptive immune
response is generated.
- Tonsils, Peyer’s patches, appendix, solitary
lymphoid follicles in the large intestine and rectum.
GALT Architecture
Lamina
Propria
** Dome structures extend into the lumen of the intestine.
Lamina Propria – effector site
Inductive Site Effector Site
Enormous Antigen Load
Systemic Immune System –
largely sterile environment. Vigorous
response to microbial invasion.
Mucosal Immune System –
Constant exposure to foreign matter
Human gut is exposed to an
enormous amount commensal
microorganisms (1 x 10 14)
Constant exposure to food matter
Innate Defense –
I. Barrier Fxn
1. Glycocalyx – Goblet cells produce mucous
to create a thick barrier that covers the GI
epithelium and prevents easy access.
- Pathogens become trapped in the mucous
and are expelled via peristalsis.
- Mucous also acts as a reservoir for
secretory IgA.
I. Barrier Fxn
Epithelial Cell Tight Junctions - prevent
the passages of macromolecules.
** Zonulin – Homology to Vibrio cholera
toxin.
upregulated during the acute phase of
celiac disease.
- Induces tight junction disassembly and
increased intestinal permeability.
Drago et al. Scand J Gastroenterol. 2006
Fasano et al. Lancet 2000
II. Proteolytic Enzymes
Enzymes in the stomach (pepsin) and small
bowel (trypsin, chymotrypsin, pancreatic
proteases).
Break down large polypeptides into
di-peptides and tri-peptides.
Peptides < 8-10 aa are poor immunogens.
Enzymes cytotoxic to pathogens.
III. Antimicrobial Molecules
1. Lactoferrin – binds iron and inhibits
bacterial growth.
2. Lysozyme – cleaves cell wall of
gram positive bacteria.
3. Defensins – 30-40 aa peptides
that disrupts the cell memebranes of
bacteria and fungi causing lysis.
IV. Commensal Organisms
>400 species of commensal bacteria
Provide enzymatic breakdown of food
Competes with pathogenic bacteria for space
and nutrients
Prevents colonization of the gut
Antibiotics disrupt homeostasis
IV. Commensal Organisms
Germ free mice – no commensal
microflora.
- Pups delivered by C-section and raised in
sterile conditions.
Hypoplastic peyer’s patches with scant
germinal centers.
- decreased IgA plasma cells
- decreased lamina propria CD4+ cells
- Abnormalities reversed by placing non-germ
free mice in same cage.
Mucosal Immune
System:
Adaptive Response
“Common Mucosal Immune
System”
*Antigen Presentation*
Peyer’s Patch
Mesenteric Lymph
Node Thoracic Duct
Blood Stream
Resp Breast Intestinal Mucosa GU Salivary/Lacrimal
Tract Tract Gland
Common Mucosal System
IgA response for different routes of vaccination
Holmgren et al. Nature Medicine. 2005
GALT – Unique Epithelium
The epithelium
overlying the peyer’s
patches is composed
of cells that differ
from the surrounding
enterocytes.
M-Cells (microfold cells)
M-cells lack microvilli
No glycocalyx coating
Designed to to interact directly
with antigens in the gut –
portal of entry into GALT.
– some pathogens gain entry
via M-cells
(salmonella, shigella)
M-Cells (microfold cells)
Basolateral aspects are
invaginated.
They contain T-cells, B-cells,
Dendritic cells, and
Macrophages.
Antigens from the lumen are
taken up by endocytosis and
presented directly to APCs
APCs migrate to germinal
center
Germinal Center
Intraepithelial Lymphocytes
Strategically located to respond to
antigenic stimulation
Most T-cells are CD8+
Mainly αβ TCR (In mice, γδ TCR
predominates).
IEL: CD8 + T-Cells
Limited Repertoire of TCR
- marked difference compared to peripheral T-
cells.
Recognize a limited # of antigens
Prevents indiscriminate inflammation
Recognition of self-stress antigens (MIC-A,
MIC-B)
- T-cells induce apoptosis of injured epithelial
cells.
Lamina Propria Lymphocytes
T-cells are predominantely CD4 +
(95% CD45RO+)
Limited capacity to proliferate
Weak proliferative responses to mitogens or
specific antigens.
Still act as helpers for B-cells
B-Cell Response: S-IgA
Secretory IgA is the predominant Ig isotype in
the gut.
Blood IgA exists mainly as a monomer
In the mucosa, IgA is exclusively dimeric
J-Chain
FUCTIONS OF sIgA
1. Protection in the sub-epithelial mucosal areas
such as upper respiratory, urogenital and
gastrointestinal tracts.
2. plays a major role in preventing antigen
uptake derived from both infectious and non-
infectious agents.
3. sIgA provides protection against infections,
allergic reactions (immediate type) and
autoimmune diseases.
4 Exclusion of bacterial and viral pathogens,
bacterial toxins and other noxious agents.
5 Clears infections through transport of pathogen
immune-IgA complexes into the bile.
6 potentiates mucous viscosity in the blocking
and removal of infectious agents.
7 mediates antibody-dependent T cell –
mediated cytotoxicity (ADCC) .
8 Interferes with utilization of growth factors like
iron by bacterial pathogens.
9 increases antigen uptake by M cells into
Peyer’s patches.
10 lymphocyte activation through anti-idiotypic
antibody stimulation.
Secretory IgA Transport
S-IgA is produced by
plasma cells in the
lamina propria.
S-IgA binds to
polymeric Ig receptor
on the basolateral
surface of intestinal
epithelial cells
Lamina Lumen
Propria
It is transported to the
intestinal lumen by
transcytosis.
Secretory IgA transport
**Secretory Component (SC) of the
receptor remains associated with IgA
SC protects IgA from proteolytic
cleavage.
SC also acts as a “glue” to bind IgA to
the glycocalyx.
IgA Subtypes
IgA 1 and IgA 2 mainly differ in their hinge
regions
IgA 1 ab contain 13 additional aa in the hinge
region.
- More flexible
- More susceptible to IgA1 specific proteases
made by bacteria.
IgA 2 is resistant to proteases
- Serum ratio 4:1
- Mucosal ratio 3:2 (even higher in colon)
Gut Anti-Inflammatory
Mechanisms: Secretory IgA
IgA is unable to activate complement by
classical or alternative pathways.
S-IgA can inhibit phagocytosis and
chemotaxis of neutrophils, macrophages
Can down regulate synthesis of
TNF-α and IL-6
T-Regulatory Cells
IPEX – severe enteropathy results from
lack of CD4+CD25+ Foxp3+ T Regs.
Naïve T-cells can differentiate into T
regs in the presence of TGF-β¹
Transfer of Tregs into mice with IBD
can lead to resolution of colitis²
1. Chen et al. Journal of Experimental Medicine. 2003.
2. Mottet et al. Journal of Immunology. 2003.
Regulatory Cytokines
IL-10 – Increased IgA
Decreased cytokine production by DC, T-
cells, macrophages
Promotes TH2 response
IL-10 knockout mice: severe enterocolitis
TGF-beta – Increased IgA
Maintain functional CD4+CD25+ cells in the
periphery.
Oral Tolerance
- Oral tolerance is the generation of systemic
immune unresponsiveness by feeding of antigen.
The antigen is usually soluble and without
adjuvant or proinflammatory activity.
- Oral tolerance is likely a mechanism for
prevention of harmful immune responses to
harmless antigens such as foods.
- A number of mechanisms may underlie oral
tolerance, including clonal deletion, clonal anergy,
or active suppression by T cells (cytotoxic, TH2, or
TGF- producing)
Oral Tolerance
- Generation of systemic immune unresponsiveness by
feeding antigens.
-Oral tolerance prevents harmful immune responses to
harmless antigens such as foods.
Features of oral tolerance
Normal immune function
Tolerance can be local or systemic
It requires a functional immune system
Symbiosis - in the absence of
commensals, a poor immune response
develops and oral tolerance cannot be
induced
Antigen specific.
Often partial (e.g. antibodies inhibited, but
T cell responses may remain).
Not complete (e.g. may be a quantitative
reduction in antibody levels).
Wanes with time.
Easier to abrogate a response than reduce
and established response.
Good immunogens are better at inducing
tolerance
Dose and route dependent.
Mechanisms
Central tolerance deletion of self-
reactive T cells in the thymus
Peripheral tolerance
1. deletion
2. immune deviation
3. anergy
4. suppression / regulation
Deletion
Central tolerance through negative selection in
the thymus by apoptosis of specific T-
lymphocytes (e.g. fas-fasL)
Shown to play a role in peripheral tolerance in
sites of immune privilege (eg stromal cells in the
testes express fasL)
orally administered antigen can induce
tolerance not only by active suppression and
clonal anergy but by extrathymic deletion of
antigen-reactive Th1 and Th2 cells
Anergy/Deletion
Immune Deviation
CD4+ T lymphocytes are activated by antigen
presenting cells (APC)
Th1 cells - important in inflammatory
responses (e.g. delayed type hypersensitivity)
Th2 cells - important in helping antibody
responses. Suppress Th1 cells (IL-4, IL-10).
Th1 immune responses may be inhibited if
Th2 cells are stimulated instead.
Inhibitory cytokines
Transforming growth factor beta (TGF)
non-specifically inhibits the growth of
lymphocytes (Th3)
Specific immune responses can be
inhibited by Th2 cytokines (IL-4 and IL-
10)
Some populations of T lymphocytes
(both CD4 and CD8) can consume IL-2,
the T cell growth factor. Surrounding
cells therefore fail to grow
Th1 cells make
gamma-interferon
CD4+ IL-12
APC T
Th2 cells make
IL-4 and IL-10
Expressing
Class II MHC Th3 cells make
TGF-beta
Anergy
Non-productive antigen presentation
T cells are activated by antigen presenting
cells
Signal absence / blockade
Some epithelial cells in the gut and lung
normally express class II MHC, but not
costimulatory molecules and therefore
cannot provide signal 2
CTLA4 Ig have been developed to block
the interaction of CD28 with B7 on APC
and therefore block signal 2
Anergy may also be due to:
Results in a specific hypresponsiveness
Anergic cells do not respond to specific
MHC+peptide plus costimulation
Anergic cells may then block APC - and
inhibit immune responses
Anergic cells may consume IL-2
Anergic cells are more susceptible to
programmed cell death (apoptosis)
Tregs
T cells specific for food antigens become Tregs and produce IL-10 and
TGF-β that inhibit other self-reactive T cells
Breakdown of oral tolerance
Immune responses to food
leadsto food intolerance
eg coeliac disease
Immune responses to commensal
bacteria
leads to inflammatory bowel disease (IBD)
eg crohn’s disease, ulcerative colitis
Tolerance to external
substances
The body is likely NOT TO REACT to:
Small, simple molecules
Oral entry
Large or very small doses
In young immunologically immature individuals
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Summary of MUCOSAL Immune defense.
Innate immunity
Physical factors
Soluble factors
Microbiological factors
Cellular factors
Adaptive immunity
T cells
Secretory IgA
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