DURING LECTURE/DISCUSSION

8/28/2019 S.Wahyuni 1
 INNATE IMMUNITY

S.Wahyuni, MD, PhD

Department of Parasitology,
Medical Faculty, Hasanuddin University
wahyunim@indosat.net.id
+62-8152531325

8/28/2019 S.Wahyuni 2
• Component of innate immunity
– Epitel cells
– Innate immunity cells
• Monocyte, macrophages, basophils, eosinophil,
dendriti cells, Natural killer cells
– Protein plasma
• Cytokines
• Complement

8/28/2019 S.Wahyuni 3
• Innate immunity serves three important
functions:
– prevents
– controls
– eliminates
infection of the host by many microbes.

8/28/2019 S.Wahyuni 4
• Innate immune mechanisms recognize
– microbes
– products of damaged
– dead host cells
– non-microbial but should not be present in
healthy tissues (intracellular crystals)

8/28/2019 S.Wahyuni 5
• Two major responses of the innate immune system:
– Inflammation:
• process by which leukocytes and circulating plasma proteins
are brought into sites of infection and activated to destroy
and eliminate the offending agents.
• reaction to damaged or dead cells and to accumulations of
abnormal substances in cells and tissues.
– antiviral defense:
• consists of changes in cells that prevent virus replication
• increase susceptibility of viral to be killed by lymphocytes

8/28/2019 S.Wahyuni 6
 Inflammation

8/28/2019 S.Wahyuni 7
 Inflammation

8/28/2019 S.Wahyuni 8
 Antiviral defence

8/28/2019 S.Wahyuni 9
 RECOGNITION OF
MICROBES AND DAMAGED SELF
BY
THE INNATE IMMUNE SYSTEM

8/28/2019 S.Wahyuni 10
 Animal properties

8/28/2019 S.Wahyuni 11
 Pathogen properties

8/28/2019 S.Wahyuni 12
• The innate immune system recognizes molecular
structures that are characteristic of microbial
pathogens but not mammalian cells.
– The microbial substances that stimulate innate
immunity are called pathogen-associated
molecular patterns (PAMPs).
– Different classes of microbes (e.g., viruses,
bacteria, fungi, parasite) express different PAMPs.

8/28/2019 S.Wahyuni 13
• The innate immune system recognizes microbial
products that are often essential for survival of the
microbes.
Pathogen-Associated Molecular Patterns Microbe Type
Nucleic acids ssRNA Virus
dsRNA Virus
CpG Virus, bacteria
Proteins Pilin Bacteria
Flagellin Bacteria
Cell wall lipids LPS Gram-negative bacteria
Lipoteichoic acid Gram-positive bacteria
Carbohydrates Mannan Fungi, bacteria
Dectin glucans Fungi

8/28/2019 S.Wahyuni 14
• The innate immune system also recognizes
endogenous molecules that are produced by or
released from damaged and dying cells.
• Called damage-associated molecular patterns
(DAMPs)
Damage-Associated Molecular Patterns
Stress-induced proteins HSPs
Crystals Monosodium urate
Nuclear proteins HMGB1

8/28/2019 S.Wahyuni 15
 Damage associate molecular pattern
8/28/2019 S.Wahyuni 16
8/28/2019 S.Wahyuni 17
• To recognize PAMPs and DAMPs, innate
immune system uses
– cellular receptors present in different
locations in cells
– soluble molecules receptors in the blood
and mucosal secretions

8/28/2019 S.Wahyuni 18
 CELL RECEPTORS
OF
INNATE IMMUNITY

8/28/2019 S.Wahyuni 19
Cell-Associated Pattern Specific
Recognition Receptors Location Examples PAMP/DAMP Ligands
Plasma membrane & TLRs 1-9 Various microbial
endosomal membranes molecules including
of dendritic cells, bacterial LPS and
phagocytes, B cells peptidoglycans, viral
endothelial cells, and nucleic acids
many other cell types
Cytoplasm of NOD1/2 Bacterial cell wall
phagocytes epithelial NALP peptidoglycans
cells, and other cells family Flagellin, muramyl
(inflammaso dipeptide, LPS; urate
mes) crystals; products of
damaged cells
RIG-like Cytoplasm of RIG-1, Viral RNA
receptors (RLRs) phagocytes and other MDA-5
cells
8/28/2019 S.Wahyuni 20
Cell-Associated
Pattern Recognition Specific
Receptors Location Examples PAMP/DAMP Ligands
C-type lectin-like Plasma Mannose Microbial surface
receptors membranes of receptor carbohydrates with
phagocytes terminal mannose and
fructose
Dectin Glucans present in fungal
cell walls
Plasma CD36 Microbial
membranes of diacylglycerides
phagocytes
N-Formyl Plasma FPR and Peptides containing N-
met-leu-phe membranes of FPRL1 formylmethionyl residues
receptors phagocytes

8/28/2019 S.Wahyuni 21
 Cellular locations of pattern recognition molecules of the innate immune system
8/28/2019 S.Wahyuni 22
 Toll like receptors

• Expressed on many cell types

• Recognize products of a wide
variety of microbes
• Also involved in response to
endogenous molecules whose
indicates cell damage.
• In humans, TLR1-TLR9

Structure, location, and specificities of mammalian TLRs

8/28/2019 S.Wahyuni 23
• Major transcription
factors signaling
pathways are:
– Nuclear factor κB (NF-
κB)
– Protein 1 (AP-1)
– Interferon response
factor 3 (IRF3) & IRF7

8/28/2019 S.Wahyuni 24
Cytosolic receptors for
PAMPs and DAMPs

• To detect infection or cell

damage in the cytoplasm
• 2 major classes receptors:
• NOD-like receptors
• RIG-like receptors
• Both receptors, are linked to
signal transduction pathways
that promote inflammation or
type I interferon production.

8/28/2019 S.Wahyuni
The inflammasome 25
Other Cell-Associated
Pattern Recognition Receptors
• Expressed on the plasma membranes of various cell
types and recognize microbial molecules
• Example:
– Receptors for Carbohydrates
– Scavenger receptors (CD36 )
– N-Formyl Met-Leu-Phe Receptors

8/28/2019 S.Wahyuni 26
 CELLULAR COMPONENTS OF THE
INNATE IMMUNE SYSTEM

Epithel, phagocytes, dendritic cells, mast cells &

natural killer cells

8/28/2019 S.Wahyuni 27
 Epithel
• Interfaces between the environment and the
mammalian host
• Are lined by continuous layers of specialized
epithelial cells
• Form tight junctions with one another,
blocking passage of microbes between the
cells.
• Loss integrity of epithelial layers: predisposes
an individual to infections.
8/28/2019 S.Wahyuni 28
• Including:
• Keratin block microbial penetration into deeper
layers of the epidermis.
• Mucus, containing glycoproteins called mucins
impairs microbial invasion and facilitates microbe
removal by ciliary action in the bronchial tree and
peristalsis in the gut.
• Antimicrobial peptide produced by epithelial
leukocytes including defensins and cathelicidins.

8/28/2019 S.Wahyuni 29
 Epithelial Barriers

8/28/2019 S.Wahyuni 30
8/28/2019 S.Wahyuni 31
 Phagocytes

• Macrophages and neutrophils

• The first line of defense against microbes that breach
epithelial barriers
• Functions
– Internalize and kill microbes
– Producing cytokines that promote inflammation
and also enhance the antimicrobial function of
host cells at the site of infection.
• Low neutrophil counts high rate of lethal bacterial
and fungal infections
8/28/2019 S.Wahyuni 32
 Dendritic Cells
• Produced by bone marrow
• Present in epithelia and most tissues of the body.
• Detect invading microbes.
• Express more different types of TLRs and cytoplasmic
pattern recognition receptors than any other cell
type, making them the most versatile sensors of
PAMPs and DAMPs among all cell types in the body.

8/28/2019 S.Wahyuni 33
• DC function:
– take up microbial protein antigens
– transport protein antigens to lymph nodes where
naive T cells home
– display protein antigens in a way that the T cells
can recognize.
– direct naive T cell differentiation into distinct
types of effector cells

8/28/2019 S.Wahyuni 34
 Mast Cells
• Present in the skin and mucosal epithelium, usually
located adjacent to blood vessels
• Have cytoplasmic granules containing various
inflammatory mediators that are released when the cells
are activated
• Vasoactive amines (such as histamine) cause
vasodilation and increased capillary permeability,
• Poteolytic enzymes  kill bacteria or inactivate
microbial toxins.

8/28/2019 S.Wahyuni 35
• Granule contents rapidly induce changes in the blood
vessels that promote acute inflammation.
• Provide defense against helminths and are
responsible for symptoms of allergic diseases.
• Synthesize and secrete lipid mediators (such as
prostaglandins) and proinflammatory cytokines (such
as TNF).
• Mast cell-deficient mice are impaired in controlling
bacterial infections

8/28/2019 S.Wahyuni 36
 Natural Killer Cells
• Arise from bone marrow precursors
• A large lymphocytes with numerous cytoplasmic granules
• 5% to 15% of the mononuclear cells in the blood and
spleen.
• Responses mainly against intracellular viruses and
bacteria.
• Perform killing function
• Use germline DNA-encoded receptors to distinguish
pathogen-infected from healthy cells.
• Distinguish infected and stressed cells from healthy cells
8/28/2019 S.Wahyuni 37
 Functions of NK cells
8/28/2019 S.Wahyuni 38
 T and B Lymphocytes with Limited Antigen
Receptor Specificities
• Certain subsets of T and B lymphocytes have very
little receptor diversity
• Recognize PAMPs
• Among other
• invariant natural killer T cells (iNKT)
• γδ T cells
• intraepithelial T cells with αβ TCRs
• B-1 B cells
• marginal zone B cells.
8/28/2019 S.Wahyuni 39
 SOLUBLE RECOGNITION AND
EFFECTOR MOLECULES OF INNATE
IMMUNITY

8/28/2019 S.Wahyuni 40
• Recognize microbes and promote innate responses
exist in soluble form in the blood and extracellular
fluids
• Also known as, ‘humoral branch of innate immunity’
• Function:
– bind to microbes, act as opsonins and enhance the
ability of macrophages, neutrophils, and dendritic
cells to phagocytose the microbes.
– promote inflammatory responses that bring more
phagocytes to sites of infections
– directly kill microbes.
• Include: collectins, pentraxins, ficolins complement
system, and natural antibodies,

8/28/2019 S.Wahyuni 41
 Soluble Recognition Location Specific Examples PAMP Ligands
Molecules
Pentraxins Plasma C-reactive protein Microbial phosphorylcholine
and phosphatidylethanolamine

Collectins Plasma Mannose-binding Carbohydrates with terminal

lectin mannose and fructose
Alveoli Surfactant proteins Various microbial structures
SP-A and SP-D
Ficolins Plasma Ficolin N-Acetylglucosamine and
lipoteichoic acid components
of the cell walls of gram-
positive bacteria
Complement Plasma C3 Microbial surfaces

Natural Plasma IgM Phosphorylcholine on bacterial

antibodies membranes and apoptotic cell
membranes
8/28/2019 S.Wahyuni 42
 The Complement System
• Several plasma proteins that work together
• Complement activation is a cascade proceeds, the enzymatic
activities result in tremendous amplification of the amount of
proteolytic products that are generated perform the
effector functions
• Function:
– Opsonize microbes
– Promote the recruitment of phagocytes to the site of
infection
– Directly kill the microbes

8/28/2019 S.Wahyuni 43
 The Complement System

• The first step in activation of the complement system

is recognition of molecules on microbial surfaces
• Has three ways, each referred to as a distinct
pathway of complement activation:
– Classical pathway
– Alternative pathway
– Lectin pathway

8/28/2019 S.Wahyuni 44
 mannose-binding lectin, and ficolin

8/28/2019 S.Wahyuni 45
 Pathways of complement activation

8/28/2019 S.Wahyuni 46
 Natural Antibodies
• Produced by subsets of B cells
• Are already present before infections
• Recognize common molecular patterns on microbes or stressed
and dying cells.
• Specific for carbohydrate or lipid molecules
• Most are IgM antibodies
• Function: protection against bacterial infections and facilitate
the phagocytosis of apoptotic cells.
• Example: anti-ABO blood group antibodies that recognize
certain glycolipids (blood group antigens) expressed on the
surface of many cell types, including blood cells.

8/28/2019 S.Wahyuni 47
 FEEDBACK MECHANISMS THAT
REGULATE INNATE IMMUNITY

8/28/2019 S.Wahyuni 48
• Function is to regulate the magnitude and
duration of innate immune responses to limit
potential damage to tissues.
• Several mechanisms have evolved to provide a
break on inflammation, and these mechanisms
come into play at the same time as or shortly
after the initiation of inflammation.
• The stimuli for the initiation of many of these
control mechanisms include the same PAMPs and
DAMPs that induce inflammation.
8/28/2019 S.Wahyuni 49
IL-10
• inhibits activation of macrophages and
dendritic cells.
• macrophages and dendritic cells produce
various inflammatory cytokines (IL-1, TNF, and
IL-12)
• Is an excellent example of a negative feedback
regulator.

8/28/2019 S.Wahyuni 50
IL-1 receptor antagonist (IL-1RA).
• Produce by mononuclear phagocytes
• Induced by many of the same stimuli that
induce IL-1 production
• Recombinant IL-1RA has been developed as a
drug that is effective in the treatment of
systemic juvenile rheumatoid arthritis and
familial fever syndromes in which IL-1
production is dysregulated.
8/28/2019 S.Wahyuni 51
Autophagy genes
• Autophagy is a mechanism by which cells
degrade their own organelles, such as
mitochondria, by sequestering them within
membrane-bound vesicles and fusing the vesicles
with lysosomes.
• This will impairs impair cytokine synthesis
• A role for Atg proteins in regulating innate
immune responses is further supported by the
discovery that polymorphisms in a human Atg are
associated with inflammatory bowel disease.
8/28/2019 S.Wahyuni 52
Suppressors of cytokine signaling (SOCS)
• proteins are inhibitors of JAK-STAT signaling
pathways linked to cytokine receptors.
• TLR signaling in macrophages and dendritic
cells induces the expression of SOCS proteins,
which limit responses of these cells to
exogenous cytokines such as type I
interferons.

8/28/2019 S.Wahyuni 53
 SUMMARY
• The innate immune system provides the first line of host defense against microbes. The
mechanisms of innate immunity exist before exposure to microbes. The cellular components of the
innate immune system include epithelial barriers, leukocytes (neutrophils, macrophages, NK cells,
lymphocytes with invariant antigen receptors, and mast cells).
• The innate immune system uses cell-associated pattern recognition receptors, present on plasma
and endosomal membranes and in the cytoplasm, to recognize structures called pathogen-
associated molecular patterns (PAMPs), which are shared by microbes, are not present on
mammalian cells, and are often essential for survival of the microbes, thus limiting the capacity of
microbes to evade detection by mutating or losing expression of these molecules. In addition, these
receptors recognize molecules made by the host but whose expression or location indicates cellular
damage; these are called damage-associated molecular patterns (DAMPs).
• TLRs, present on the cell surface and in endosomes, are the most important family of pattern
recognition receptors, recognizing a wide variety of ligands, including bacterial cell wall
components and microbial nucleic acids. Cytoplasmic pattern recognition receptors exist that
recognize microbial molecules. These receptors include the RIG-like receptors (RLRs), which
recognize viral RNA, and the NOD-like receptors (NLRs), which recognize bacterial cell wall
constituents and also sense sodium urate and other crystals.
• Pattern recognition receptors, including TLRs and RLRs, signal to activate the transcription factors
NF-κB and AP-1, which promote inflammatory gene expression, and the IRF transcription factors
that promote expression of the antiviral type I interferon genes. The inflammasome, a specialized
complex that forms in response to PAMPs and DAMPs, is

8/28/2019 S.Wahyuni 54
• Pattern recognition receptors, including TLRs and RLRs, signal to activate the transcription factors NF-κB and AP-1,
which promote inflammatory gene expression, and the IRF transcription factors that promote expression of the
antiviral type I interferon genes. The inflammasome, a specialized complex that forms in response to PAMPs and
DAMPs, is composed of a NOD-like receptor, an adaptor, and the enzyme caspase-1, the main function of which is
to produce active forms of the inflammatory cytokines IL-1 and IL-18.
• Soluble pattern recognition and effector molecules are found in the plasma, including pentraxins (e.g., CRP),
collectins (e.g., MBL), and ficolins. These molecules bind microbial ligands and enhance clearance by complement-
dependent and complement-independent mechanisms.
• NK cells are lymphocytes that defend against intracellular microbes by killing infected cells and providing a source
of the macrophage-activating cytokine IFN-γ. NK cell recognition of infected cells is regulated by a combination of
activating and inhibitory receptors. Inhibitory receptors recognize class I MHC molecules, because of which NK
cells do not kill normal host cells but do kill cells in which class I MHC expression is reduced, such as virus-infected
cells.
• The complement system includes several plasma proteins that become activated in sequence by proteolytic
cleavage to generate fragments of the C3 and C5 proteins, which promote inflammation, or opsonize and promote
phagocytosis of microbes. Complement activation also generates membrane pores that kill some types of
bacteria. The complement system is activated on microbial surfaces and not on normal host cells because
microbes lack regulatory proteins that inhibit complement. In innate immune responses, complement is activated
mainly spontaneously on microbial cell surfaces and by mannose-binding lectin to initiate the alternative and
lectin pathways, respectively.
• The two major effector functions of innate immunity are to induce inflammation, which involves the delivery of
microbe-killing leukocytes and soluble effector molecules from blood into tissues, and to block viral infection of
cells by the antiviral actions of type 1 interferons. Both types of effector mechanism are induced by the PAMPs
and DAMPs, which initiate signaling pathways in tissue cells and leukocytes that activate transcription factors and
lead to the expression of cytokines and other inflammatory mediators.

8/28/2019 S.Wahyuni 55
• Several cytokines produced mainly by activated macrophages mediate inflammation. TNF and IL-1 activate
endothelial cells, stimulate chemokine production, and increase neutrophil production by the bone marrow. IL-1
and TNF both induce IL-6 production, and all three cytokines mediate systemic effects, including fever and acute-
phase protein synthesis by the liver. IL-12 and IL-18 stimulate production of the macrophage-activating cytokine
IFN-γ by NK cells and T cells. These cytokines function in innate immune responses to different classes of
microbes, and some (IL-1, IL-6, IL-12, IL-18) modify adaptive immune responses that follow the innate immune
response.
• Neutrophils and monocytes (the precursors of tissue macrophages) migrate from blood into inflammatory sites
during innate immune responses because of the effects of cytokines and chemokines produced by PAMP- and
DAMP-stimulated tissue cells.
• Neutrophils and macrophages phagocytose microbes and kill them by producing ROS, nitric oxide, and enzymes in
phagolysosomes. Macrophages also produce cytokines that stimulate inflammation and promote tissue
remodeling at sites of infection. Phagocytes recognize and respond to microbial products by several different types
of receptors, including TLRs, C-type lectins, scavenger receptors, and N-formyl met-leu-phe receptors.
• Molecules produced during innate immune responses stimulate adaptive immunity and influence the nature of
adaptive immune responses. Dendritic cells activated by microbes produce cytokines and costimulators that
enhance T cell activation and differentiation into effector T cells. Complement fragments generated by the
alternative pathway provide second signals for B cell activation and antibody production.
• Innate immune responses are regulated negative feedback mechanisms that limit potential damage to tissues. IL-
10 is a cytokine that is produced by and inhibits activation of macrophages and dendritic cells. Inflammatory
cytokine secretion is regulated by autophagy gene products. Negative signaling pathways block the activating
signals generated by pattern recognition receptors and inflammatory cytokines.

8/28/2019 S.Wahyuni 56