Innate Immunity

Hathairat Thananchai, DPhil

Department of Microbiology
Faculty of Medicine
Chiang Mai University
2 August 2016

Objectives:
Explain how innate immune system recognizes foreign
substances
Explain components of the innate immune system,
and their functions
Explain functions of the innate immune system

Lecture outline
Overview of innate immunity
Innate recognition
Cell-associated receptors
Soluble molecules

Cellular and soluble components of innate immunity

Innate immune responses
- Inflammatory response
- antiviral response
Stimulation of adaptive immunity

Innate and adaptive immunity

The mechanisms of innate immunity provide the initial defense against

infection. Adaptive immune responses develop later and consist of activation
of lymphocytes.

Innate immunity
Innate immunity serves three important functions
Innate immunity is initial response to microbes that prevents,
controls , or eliminate infection of the host by many microbes
Inflammation
Antiviral defense

Innate immune mechanisms recognize the products of

damaged and dead host cells and serve to eliminate these
cells and to initiate the process of tissue repair
Innate immunity to microbes stimulates adaptive immune
responses and can influence the nature of the adaptive
responses to make them optimally effective against different
types of microbe

Features of Innate immunity

Provides the early line of defense against foreign
substances
Not specific to particular microbe
do express surface receptors that distinguish host cells
from those of infectious agents
do not discriminate between various infectious agents

Non-adaptive
the quality of the reaction to a foreign substance does
not change when the organism encounters this
substance repeatedly

Recognition mechanisms of innate immunity

The microbial substances that stimulate innate immunity are
called pathogen-associated molecular patterns (PAMPs)

Molecules recognized tend to be structural elements that

are common to broad classes of microbes and are very hard
to change.
The innate immune system also recognizes endogenous
molecules called damage-associated molecular patterns
(DAMPs)
The receptors that bind to PAMPs or DAMPs are called
pattern recognition receptors (PRRs)

Innate immune recognition of bacterial cell wall components

DAMPs

Examples of PAMPs and DAMPs

Specificity of innate and adaptive immunity

Pattern Recognition Molecules of the Innate Immune System

Cell-associated molecules
Cell membrane
Cytoplasm
Endosome

Soluble molecules
Blood
Extracellular fluids
Recruitment and activation of
protein kinases
Activation of transcription factors
Gene transcription

Innate cytokines include

- TNF
- IL-1, IL-6, IL-10, IL-12
- Type I interferons

Pattern Recognition Molecules of the Innate Immune System

Toll-like receptors (TLRs)

A family of conserved cellular
receptors that mediated
cellular responses to PAMPs
and DAMPs
TLRs activation is essential for
provoking the innate response
and enhancing adaptive
immunity against pathogens

There are 9 different functional

TLRs in humans, named TLR1
through TLR9
TIR; TLR/IL-1 receptor

TLR signaling
All TLRs except TLR3 signal through
adaptor protein MyD88
TLR3 signals through TRIF
TLR4 signals through both MyD88
and TRIF
A downstream effect of TLR
signaling through MyD88 is the
activation of the transcription
factor NF-B
A downstream effect of TLR
signaling through TRIF is the
activation of IRF-3 and -7
MyD88; myeloid differentiation primary response gene 88

TIRF; TIR domain-containing adaptor inducing IFN-

IRF; interferon response factor

Cytosolic pattern recognition molecules

NOD-Like Receptors
NOD-like receptors (NLRs) are a family
of more than 20 different cytosolic
proteins, some of which recognize
PAMPs and DAMPs
NOD1 & NOD2 recognize
peptidoglycan substructures and
promote innate immune responses
NOD1 diaminopimelic acid (DAP)
NOD2 muramyl dipeptide
Induce the production of
proinflammatory cytokines (IL-1 and
IL-18)
NOD; nucleotide oligomerization domain

RIG-Like Receptors
RIG-like receptors (RLRs) are
cytosolic sensors of viral RNA
that respond to viral nucleic acid
by inducing the production of
the antiviral type I interferons
The two best characterized RLRs
are
RIG-1
MDA5
On binding viral RNA, the RLRs
initiate signaling events that lead
to phosphorylation and
activation of IRF3 and IRF7, as
well as NF-B

Plasma membrane

Cytosolic DNA Sensors (CDSs)

The STING (Stimulator of IFN Genes)
pathway is a major mechanism of
DNA-induced activation of type I
interferon responses
STING is an endoplasmic reticulumlocalized transmembrane protein
Cytosolic DNA binds to an enzyme
called cyclic GMP-AMP synthase
(cGAS) that synthesizes a cyclic
dinucleotide called cyclic GMP-AMP
(cGAMP)
cGAMP interacts with and stimulates
STING

The differential expression of PRRs by some immune cells

The components of the innate immune system

Physical barriers
Skin and mucosal surface
Cellular components
Phagocytes (neutrophils, macrophages)
Dendritic cells
NK cells and other innate lymphoid cells
Innate-like lymphocytes
Mast cells

Circulating proteins
Complement system
Antimicrobial peptides
Cytokines : inflammation (TNF, IL-1)
chemokines (IL-8, MCP-1)
anti-viral (type I interferons)

Exterior defenses
Most infectious agents are prevented from entering the body by physical and
biochemical barriers. The body tolerates a number of commensal organisms,
which compete effectively with many potential pathogens.

Epithelial defenses

Antimicrobial peptides
Short, cationic peptides (most 29-35
amino acids long)

Made by neutrophils and epithelial cells

(small intestines, respiratory tract,
genitourinary tract)
In human, three major groups of these
antimicrobial peptides are recognized :
- -defensins
- -defensins
- cathelicidins
Differentially active against different
microorganisms

Biological roles of host defense peptide

Nature Biotechnology 2006;24:1551-1557.

Pathogens and Disease 2014;70:257-270.

Phagocytes

Neutrophil

Mononuclear phagocyte

Dendritic cell

- Phagocytosis
- Cytokine production :
Pro-inflammatory cytokines (TNF, IL-1)

Phases of phagocytosis

Phagocytosis and intracellular destruction of microbes

NADPH oxidase

Killing of phagocytosed microbes

Reactive oxygen species (ROS)
- Phagocyte oxidase (=NADPH oxidase)
superoxide anion (O2- )
- Superoxide dismutase H2O2
- Myeloperoxidase hypohalous acid
The process by which ROS are produced
is called the respiratory burst
Reactive nitrogen intermediates,
mainly nitric oxide (NO)
- Inducible Nitric oxide synthase (iNOS)
Chronic granulomatous disease (CGD):
genetic defect in phagocyte oxidase components
(most commonly gp91, which is X-linked)

Antibody-mediated opsonization and phagocytosis of microbes

opsonin

Neutrophils

The most abundant population of

circulating white blood cells and mediate the
earliest phases of inflammatory responses.
The nucleus of a neutrophil is segmented
into 3-5 connected lobules.
The cytoplasm contains granules of two
types.
- Specific granules are filled with
enzymes such as lysozyme, NADPH
oxidase
- Azurophilic granules are lysosomes
containing enzymes ([Link])
and other microbicidal substances
([Link])

Mononuclear phagocyte system

The mononuclear phagocyte system includes circulating cells
called monocytes and tissue resident cells called macrophages

Maturation of mononuclear phagocytes

Effector functions of macrophages

Dendritic cells
A heterogeneous family of cells with long dendrite-like cytoplasmic processes
Constitutively present in lymphoid tissues, mucosal epithelium, and organ
parenchyma
Dendritic cells express more different types of TLRs and cytoplasmic PRRs
than any other cell types
Dendritic cells serve a critical function in adaptive immune responses by
capturing and displaying microbial antigen to T lymphocytes.
TLR signaling induces dendritic cell expression of costimulators and cytokines
that are needed for the activation of nave T cells and their differentiation to
effector T cells

Antigen presenting cells

Natural killer (NK) cells

A lineage of cells related to lymphocytes that recognize infected and/or
stressed cells
Kill various target cells without a need for additional activation
NK cells are a major source of IFN-
The expansion and activity of NK cells are also stimulated by cytokine,
mainly IL-15 and IL-12
High concentration of IL-2 also stimulate the activities of NK cells

[Link]

NK cells are early component of the host response to viral

infection

Functions of NK cells
1. NK cells recognize ligands on infected cells or cells undergoing
other types of stress, and kill the host cells.

The NK cells are activated and kill the antibody-coated cells.

NK cells bind to antibody-coated cells by Fc receptors and destroy these

cells. These process is called antibody-dependent cell-mediated cytotoxicity

(ADCC)

Functions of NK cells
2. NK cells respond to IL-12 produced by macrophages and secrete
IFN-, which activates the macrophages to kill phagocytosed microbes.

Innate lymphoid cells

The three subsets of ILCs produce
different sets of cytokines, participate
in host defense against distinct
pathogens, and different inflammatory
disorders
These subsets are analogous to the
TH1, TH2 and TH17 subsets of CD4+T
lymphocytes that secrete some of the
same cytokines

Group 1 ILCs produce IFN- include NK

cells
Group 2 ILCs, like TH2, secrete IL-5, IL-9
and IL-13
Group 3 ILCs are found at mucosal sites
and produce IL-17 and IL-22

The three main classes of innate-like lymphocytes and their properties

Mast cells
present in the skin and mucosal epithelium
Mast cells express high-affinity Fc receptors
for IgE

The granules in mast cells contain vasoactive

amine (i.e. Histamine), and proteolytic
enzymes that can kill bacteria or inactivate
microbial toxins
Mast cells also synthesize and secrete lipid
mediators (i.e prostaglandins) and cytokines
(i.e TNF)

Mast cells express TLRs, and TLR ligands can

induce mast cell degranulation
Play a role in allergic reaction and helminthic
infections

Complement system
Consists of about 30 serum and membrane proteins that can
mediate a variety of immune reactions
The active components of complement are generated from
inactive precursors by a cascade of proteolytic reactions
These can be triggered in any of three ways:
Lectin pathway
Classical pathway
Alternative pathway

opsonization

Innate recognition by soluble collectins and ficolins

Soluble proteins present at mucosal surfaces and/or in the bloodstream
Recognize distinctive carbohydrate configurations that occur on the
surfaces of microbes
In human, three collectins and three ficolins are known to participate in
immunity

Collectins:
- surfactant protein A (SP-A)
- surfactant protein D (SP-D)
- mannose-binding lectin (MBL)

Ficolins:
- L- , H- , and M-ficolin

C1q, mannose binding lectin, and ficolin can initiate complement activation on
binding to their ligands on cell surfaces

Pentraxins
pentameric plasma proteins

belong to the pentraxin family

short pentraxin
- C reactive protein (CRP)
- Serum amyloid P (SAP)
long pentraxin
- Pentexin 3 (PTX3)
recognize : microbes (bacteria and fungi)
apoptotic cells
functions
- Opsonin
- Activation of complement by binding to C1q
and initiating classical pathway

Pentraxins biosynthesis

Acute-phase reactants

Nat Clin Pract Rheumatol 2006 2:481-90.

The Inflammatory Response

Acute inflammation is a major way by which the innate immune system
deals with infections and tissue injury
Accumulation of leukocytes, plasma proteins, and fluid derived from the
blood at an extracellular tissue site of infection or injury
to kill microbes and begin to repair tissue damage
Acute inflammation can develop in minutes to hours and last for days

If the infection is not eliminated or the tissue injury is prolonged

chronic inflammation
Chronic inflammation sites
fibrosis

tissue remodeling, with angiogenesis and

Adaptive immunity may also be involved because cytokines produced by

T cells are powerful inducers of inflammation

Innate immune initiation of inflammation

Cell 2010;140:771-776

- Vasodilation :

blood flow

vascular permeability : leaks of plasma proteins (complement

proteins, antibodies, acute-phase reactant)

adhesiveness of circulating leukocytes to the endothelial lining

of venules : leads to accumulation of inflammatory cells

Leukocyte recruitment to sites of inflammation

diapedesis

Important cytokines and

chemokines secreted by
macrophages and dendritic
cells in response to PAMPs
or DAMPs

Local and systemic actions of cytokines in inflammation

The Antivirus Response

Type I interferon : mediate the early innate immune response
to viral infection
IFN- : Plasmacytoid dendritic cell
Mononuclear phagocytes
IFN- : many cells such as fibroblast
The most potent stimuli for type I interferon synthesis are viral
nucleic acids
Recognize by - RIG-Like receptors
- DNA sensors in the cytosol
- TLR 3, 7, 8, and 9 in endosomal vesicle

Mechanisms of induction of type I interferons by viruses

Biological actions of type I interferons

Biologic actions of type I IFNs

Inhibits viral replication

Increases expression of MHC class I
molecules
Stimulates the development of TH1
cells

Increases the cytolytic activity of NK

cells

Role of innate immunity in stimulation adaptive

immune responses
The innate immune response
provides signals that function in
concert with antigen to stimulate
the proliferation and differentiation
of antigen-specific T and B
lymphocytes
The signals generated during innate
immune responses to different
microbes influence the nature of
the adaptive response

Three kinds of signals are involved

in activation of nave T cells by
antigen-presenting cells
Signal 1 : antigen-specific signals derived
from the interaction of a specific
peptide:MHC complex with the T-cell
receptor
Signal 2 : the co-stimulatory signals that
are primarily involved in promoting the
survival and expansion of the T cells
Signal 3 : delivered by antigen-presenting
cell, which are primarily involved in
directing T- cell differentiation into the
different subsets of effector T cells

Development of TH17 subset of CD4+T cells

Summary
The innate immune system provides the first line of host
defense against microbes
The innate immune system used pattern recognition
receptors to recognize structures called PAMPs and DAMPs

Cell-associated PRRs signal to activate the transcription factor

: NF-B, AP-1
inflammatory cytokines expression
IRF transcription factors
type I interferon expression
Soluble PRRs and effector molecules bind microbial ligands
and enhance clearance by complement-independent and
complement dependent mechanisms

Summary
The two major types of responses of the innate immune
system that protect against microbes are inflammation and
antiviral defense
Molecules produced during innate immune responses
stimulate adaptive immunity and influence the nature of
adaptive immune responses