Mucosal Immunology

Parameth Thiennimitr M.D., Ph.D.

Department of Microbiology
Faculty of Medicine, Chiang Mai University

2016

Learning Objectives
1. Unique features of mucosal immune system

2. Organization of mucosal immune system

(induction, trafficking and effector sites)

3. Gut immune system & gut microbiota

4. Nasopharygeal-oral immune system
5. Mucosal vaccine and mucosal (oral) tolerance

Mucosal Immune System

MALTs = Mucosal-Associated Lymphoid Tissues

Colonized with
beneficial microbes
microbiota

NALT
Aerodigestive
tract

(Nasal-associated
lymphoid tissue)

Continuously expose to
environment
food

pathogen

Gastrointestinal
(GI) tract

Different immune
responses to
different antigens

Genitourinary
tract
pathogen

food
tolerance

inflammation

GALT
(Gut-associated
lymphoid tissue)

Human Microbiome

Human Microbiota & Immune Systems

Systemic immune system

Mucosal immune system

Human
microbiota

Organization of Mucosal Immune System

(1) Inductive Site

Inhaled
antigen

Ingested
antigen

NALT

GALT (Peyers patch)

epithelium

gut epithelium

Homing
molecules

nasal lamina
propria

gut lamina
propria

(2) Trafficking Site

Regional lymph
node

Lymphatic vessel

Blood vessel

(3) Effector Site(s)

Salivary gland

Bronchi

Mammary gland

Small bowel

Large bowel

Genital tract

Gut Immune System

Lamina propria

Gut epithelial layer

Mucus
Gut lumen

1. By M-cell

2. By dendrite of DC

Gut luminal antigen uptake

Mucosal tolerance or activation

3. By IgG

Gut Immune System

(1) Inductive Site
- GALT
- Mesenteric lymph
node
GALT
(Peyers patch)

Mesenteric
lymph node

(2) Lymphocyte
Trafficking
-

Lymphatic vessels

Blood vessels

Lymphatic &
Blood vessels

(3) Effector Site

- Gut epithelium
- Lamina propria

Normal gut
mucosa

(1) Peyers Patch:

The Gut Immune System Inductive Site
Follicle-associated epithelium (FAE)
- M (membrane or Microfold) cells sample
luminal antigens directly

B cell follicles
- B cells
- Follicular dendritic cell
(FDC)
- Follicular helper T cell

- Most of B cells are

surface IgA+

Interfollicular T-cell
areas

- No afferent
lymphatic vessel
- No capsule

Follicle-Associated Epithelium (FAE)

2. Less mucus and
antimicrobial productions

1. Pathogen entrance

Follicle-associated
epithelium (FAE)

M
cells

Subepithelial
dome

Lymphoid
follicle

Follicle-Associated Epithelium as
the Entrance of Luminal Antigens
Bacterial
adhesion/invasion

TLRs

Toxin

M cells

Chemokines:
CCL9, CCL20

Some pathogens use M cells to enter

gut immune system

M (microfold) cell

Enteric bacteria
Salmonella uses M cell
as a port of entry

HIV-1 and Reovirus

attach to M cells

(2) Lymphocyte Trafficking (Homing)

Inductive site

Effector site
- Gut epithelium
- Gut lamina propria

Cells without homing molecules

cannot get out to effector site

Lymphocyte
Homing
Lymphocytes leave
GALT via draining
Afferent
lymphatic vessel

Mesenteric
lymph node

Efferent
lymphatic
vessel

Lamina
propria
venule
endothelium

Blood
system

Thoracic duct

Lymphocyte Trafficking

Effector site

Blood
system

Trafficking
and homing

Inductive site

Lymphocyte Trafficking from Gut is Navigated by

Homing Molecules

Activated B cells from GALT

express different
homing molecules
(47, 41, CCR9, CCR10)

Inductive site
GALT

Different
Effector sites
- Secrete different
cytokines to attract
different lymphocytes
Small bowel epithelium:
MAdCAM-1 targets 47
CCL25 attracts CCR9

Large bowel epithelium:

MAdCAM-1 targets 47
CCL28 attracts CCR10

(3) The Gut Immune System Effector Site:

Normal Gut Mucosa

Gut lumen

Gut epithelium

Intestinal
(mucosal) barrier

Gut lamina
propria

Gut immune cells

(B, T and DC cells)

Gut Mucosal Barrier

Biological (gut microbiota)
- Colonization resistance
- Competition for food and space
- Stimulate gut immune development

Mucus layer

Chemical:
defensin

IgA

- Mucin from Goblet cell

- Defensin from Paneths
cell
- IgA

mucus

Paneth
cell

Goblet
cell

Gut epithelium

Physical:
- Tight junction
proteins

Intestinal
(mucosal)
barrier

Gut Epithelium

Tight junction protein

Strengthen gut barrier

Intraepithelial lymphocyte (IEL)

- Exclusively T cell

- Abundance of T cell
- Display cytotoxic phenotype

Gut epithelial innate defense

Goblet cell
- mucus

Paneth Cell
- Antimicrobial peptides (defensin)

Gut Epithelial Pattern Recognition Receptor

Gut epithelium contains TLRs, NLRs and RLRs

Secretory IgA (SIgA) Dominates in Mucosa

(except lower respiratory tract and genital tract)

Lacrimal glands

Nasal mucosa

IgA

IgA
Salivary glands

Respiratory tract

IgA

IgG
Mammary glands

Small intestine

IgA

IgA
Large intestine

IgA

Genital tract

IgG

Rationale of Mucosal Secretory IgA (SIgA)

1. Localized at the entrance of pathogen
(mucus layer)
2. Ag-specificity
independent

SIgA

Gut lumen

(carbohydrate part bind to

pathogen adhesion molecules)

mucus

3. Unusually
cross-reactive
(bind to a broader range of antigens)

Gut epithelium

4. Poor complement activator

(less inflammation)

6. Homeostasis between gut

microbiota & immune cells
5. Highly resistant to cleavage
by proteases

Secretory IgA play many roles in gut immune system

Neutralize
pathogen and
toxin in lumen

Epithelial cell

Lamina propria

Neutralize
pathogen and
toxin in
endosome

Secrete
pathogen and
toxin from
lamina propria

Modifies innate
immune
functions in
lamina propria
Inhibit
Complement & NK
cell
Promote
Opsonization &
Eosinophil
degranulation

Gut Microbiota a Group of Beneficial Microbes

Living in Gastrointestinal Tract

Balanced Gut Microbiota:

Gastrointestinal Tract

4 Dominant Bacterial Phyla

The highest recorded of any microbial habitat

(Whitman et al., 1998)

Obligate anaerobes

Colon: 1011-1012 cells/ml

Firmicutes
Bacteroi
-detes

Proteobacteria
Facultative anaerobes
Adapted from Winter SE. and Bumler AJ, Cellular Microbiology, 2014

Actinobacteria

Beneficial Roles of Gut Microbiota

Gut lumen

pathogens

1. Colonization resistance

Gut
microbiota

Dietary fiber

Short chain
fatty acid

Gut epithelium

3. Maintaining
intestinal
barrier
integrity

2. Regulation of gut
immune homeostasis

Gut lamina propria

Immune cells

4. Providing host
some nutrients

Antibiotic Disrupts Gut Immune System

Gut microbiota
colonization in
GI tract

Antibiotic kill
gut microbiota

Pathogen
overgrow

Inflammatory
response

Aberrant Gut Immune Response &

Human Diseases

Imbalanced Immune System and Metabolic Syndrome

Metabolic syndrome (MS)

MS
Systemic
inflammation

Obesity
Insulin resistance
Dyslipidemia
Hypertension
Hyperglycemia

Systemic inflammation
-

Unknown
molecular origin ?
High fat diet consumption

Increased plasma
lipopolysaccharide (LPS),
reactive oxygen species
(ROS) levels

Imbalanced Gut Microbiota & Gut Immune System

found in an Obesity
Lean

Obesity

Imbalanced gut
immune cells

Balanced gut
immune cells

(more pro-inflammatory
cytokines: IFN-, IL-17)

(more anti-inflammatory
cytokines:IL-10, IL-22)

Balanced gut microbiota

more diverse and less in
LPS-containing bacteria

Imbalanced gut microbiota

(gut dysbiosis)
Less diverse and more
LPS-containing bacteria

High fat diet

Gut lamina
propria

Gut epithelium

Gut lumen

LPS = lipopolysaccharide
ILC = Innate lymphoid cell

Nasopharyngeal and
Oral Immune System

Inductive sites of the respiratory tract

NALT

NALT =
Nasopharynxassociated lymphoid
tissues
BALT =
Bronchus-associated
lymphoid tissues

Nasopharyngeal and Oral Mucosal Immune System

(1) Inductive Site
- NALT
- Cervical
lymph node

Nasal
mucosa

Cervical
LN.

(2) Lymphocyte
Trafficking or
Homing

(3) Effector Site

- Nasal passages
- Submandibular
glands

Nasal
passage

Submandibular gland

(1) Palatine Tonsil:

The Nasopharyngeal-Oral Immune System Inductive Site

- Non-encapsulated
- Epithelium: M cells
- Follicular B cell zone:
Palatine tonsil

IgG > IgM = IgA

nave & memory T cells
- Interfollicular T cell

zone:
CD4+ > CD8+

Dendritic cells

(2) Lymphocyte Trafficking Navigated by Homing Molecules

Inductive site
NALT

Activated B cell from NALT that

express different

homing molecules

(41, CCR7, CD62L, CCR10)

Effector sites
Lacrimal, Nasal,
Salivary Glands

(3) Effector site: Nasal Passage (Nasal Lamina Propria)

Nasal passage (nasal lamina propria)

Inhaled antigen
Uptake by

M-cells or DCs in
nasal epithelium

cervical lymph nodes

Antigen specific T & B

cells homing to

nasal passage

TH2 cytokines
(IL-4, IL-5) for IgA
antibody production

(3) Effector site: Submandibular gland

Inhaled antigen

IgA

Uptake by

M-cells or DCs in
nasal epithelium

cervical lymph nodes

Antigen specific T & B

cells homing to

Submandibular gland

IgA class switch

recombination

TH2 cytokines
(IL-4, IL-5) for IgA
antibody production

Oral cavity is protected by both

Mucosal and Systemic Immunity
Mucosal Immunity:

SIgA in Saliva

Systemic Immunity:
IgG in gingival
crevicular fluid

Immune responses to Subgingival Microbiota

in Periodontal Disease Pathogenesis
Periodontal disease:
Healthy gingiva:
No inflammatory cells

Subgingival Microbes + oral innate immunity =

Inflammation
Then, stimulate adaptive immunity

Mucosal Vaccine

The Organization of MALT relevant to

Mucosal Vaccination

NALT

Antigen uptake at inductive

site of GALT and NALT

DCs at subepithelial dome process

and present Ag to T & B cells

Activated T & B cells go to regional

lymph node (mesenteric and cervical)

Circulation

Distributed widely to distinct

mucosal effector sites

GALT

Advantages of Mucosal Immunity in Vaccination

Vaccine
route

Mucosal IgA production

Oral

small intestine, ascending

colon, stomach and mammary
gland

Nasal

upper airway, saliva, nasal

secretion, and cervico-vaginal
mucosa. (not gut mucosa)

Rectal

rectum and sigmoid colon

Intravaginal

female genital tract

Nasal route immunization activates

Cervico-Vaginal immunity

Mucosal (Oral) Tolerance

Mucosal Tolerance
A state of reduced immunologic
responsiveness to an antigen induced
by mucosal exposure
e.g. oral (feeding)

Mechanisms of Mucosal Tolerance

1. Clonal deletion
Oral
exposure to
high dose
antigen

Ag-reactive T cell

2. Clonal anergy

Ag-reactive T cell

Oral repeated
exposure to
low dose
antigen

Anergic T cell

Induction of Treg cell

Ag-reactive T cell

Treg cell (regulatory T cell)

Mechanisms of Mucosal Tolerance

(1) MALT epithelium:
Secrete cytokines that promote
Treg cells shortly after mucosal
exposure

(2) Mesenteric lymph

nodes:
Tolerogenic (CD103+)
dendritic cells take up antigens

Treg cells

then induce Treg cells

(3) Systemic lymphoid

sites:
Large dose of mucosal antigens
reach systemic lymphoid site
(spleen)

spleen
mesenteric L.N.

Mucosal Tolerance Inhibits T cell Response

Inhibited Effector
Cells

Sensitivity to
Mucosal Tolerance

TH1 cells

Most

TH2 cells

Immediate

B cells

Refractory

T-cell dependent
antigen

Good induction of
mucosal tolerance

e.g. protein

T-cell independent
antigen
e.g. polysaccharides

Poor induction of
mucosal tolerance

1. Phillip D. Smith. et. al, Principles of Mucosal Immunology, 1st ed. New
York: Garland Science; 2013

2. Parham P. The Immune System. 4th ed. New York: Garland Science; 2015
3. Tak W. Mak, Marry E. Saunders and Bradley D. Jett, Primer to the immune
response, 2nd ed. Elsevier; 2014

5. Luck Helen, Sue Tsai, Jason Chung, Xavier Clemente-Casares, et al. (2015)
Regulation of obesity-related insulin resistance with gut anti-inflammatory
agents. Cell Metab 21:527-542.