Natural (Innate) Immunity

Elements of Innate Immunity

Dr. Nahla Melake

Pathology Department- Immunology section - College of Medicine – Al-Imam
Mohammad Ibn Saud Islamic University
 Objectives
✓ Recognize the significance of the immune system in defense against
infection and disease
✓ Distinguish between non-specific (innate) and specific (adaptive) immune
systems
✓ Describe the mechanisms combating infection/disease (killing pathogens)
✓ Define the humoral and cellular components of the innate immunity
✓ Recall the mechanism of action of the humoral and cellular components of
innate immunity.
✓ Describe in details opsonisation and phagocytosis process
 Body defense against
Immunity exogenous (microbes) and
endogenous (tumor cells)
agents.
Two Types of Immunity

Innate Immunity • First line of immune response

- natural, • Inborn resistance, genetically determined
- non-adaptive, or • No need for prior exposure
- non-specific • Not modified significantly by repeated exposure

• Response if innate immunity fails

Acquired Immunity • Not genetically determined
- adaptive, or • Develops during lifetime
- specific • May be acquired naturally or artificially
• Modified by repeated exposure
 Innate immunity
▪ Based on genetic make-up (Already formed components)
▪ Rapid response: within minutes of infection (0-4 hrs)
▪ Not specific (Same molecules / cells respond to a range of pathogens)
▪ Has no memory (Same response after repeated exposure)
▪ No clonal expansion of Ag specificity
▪ Failure ==> adaptive immune response
Mechanism of Innate immunity:

First line Second line

Anatomical barriers

1) Mechanical factors Humoral components Cellular components

2) Chemical barriers
3) Biological factors: ▪ Acute phase proteins ▪ Neutrophils
▪ Lysozymes ▪ Monocytes
Normal bacterial flora ▪ Macrophages
▪ Complement
▪ Cytokines ▪ NK cells
▪ Eosinophils
▪ Interferons
 First line
Mechanical barriers
▪ Intact skin and mucous membrane
▪ The hair in the nares, coughing and sneezing
▪ Mucous secretions, trap many organisms
▪ Movement of cilia in respiratory tract push out any foreign organisms
▪ The blinking reflex and tears expel any foreign organisms

Chemical barriers
▪ Sweat and sebaceous secretions (high fatty acids, acidic PH)
▪ Hydrolytic enzymes (saliva), HCL (stomach), proteolytic enzymes (small
intestine)
▪ Lysozymes (skin, tears, cervical secretions) break down peptidoglycan of cell
wall.
▪ Acidic pH in adult vagina Defensins
▪ Low molecular weight proteins in lung and
▪ Surfactants in the lungs act as opsonins GIT having antimicrobial activity
(promote phagocytosis) ▪ Granules in neutrophils are the richest
source which disrupt biological membranes.
▪ Defensins in skin and cryptocidins in GIT
 Biological barriers: Normal bacterial flora
Are non-pathogenic or weakly pathogenic bacteria and fungi that prevents
colonisation of body surface with pathogenic organisms by:
1. competition for attachment sites,
2. competition for essential nutrients or
3. production of inhibitory substance as colicins or acids

Second line
Circulating effector proteins
➢ Acute phase proteins: ✓C-reactive protein (CRP),
✓Fibrinogen,
✓Serum amyloid A protein,
▪ Synthesis in liver ✓Mannose binding lectins.
▪ Normally present in trace levels in serum
▪ Early acute inflammation indicator:
o increases within 4-6 hrs of infection or trauma
o 100 to 1000 fold increase serum concentration
o concentration drops rapidly in serum when stimulus removed
 ▪ Function: ▪ Enhances opsonization,
▪ Enhances agglutination,
▪ Enhances precipitation,
▪ Classical pathway of complement activation
▪ Enhances removal of irritant
➢ Lysozymes

• Are enzymes that damage bacterial cell walls by attacking peptidoglycans

(especially Gram +ve bacteria)
• Abundant in secretions (tears, saliva, human milk, and mucus)
• Also present in cytoplasmic granules of macrophages and PMNs

➢ Complement

• Series of  30 plasma (serum) proteins, activated in a cascade

• 5-10% of the total serum proteins
• After activation, they have enzymatic activity
Complement pathways:

Spontaneous hydrolysis
on pathogen surface
Complement components
C1 (C1q, C1r, C1s) Factor B •
C2 (C2a, C2b) Factor D •

CF= Complement Factor

C3 (C3a, C3b) DAF, CD55 •
MBL-associated serine proteases
C4 (C4a, C4b) CR1, CD35 •
C5 (C5a, C5b) Factor H •
mannose-binding lectin C6 Factor I •
C7 •
C8 •
C9 •

C3 Plays a central role in

complement activation

MAC = Membrane Attack

Complex:

o C5b678 (9)n , 12-15 C9

o Size: 10-11 nm inner diameter
o Forms a channel through the
membrane enabling ions and
small molecules to diffuse
freely across the membrane
Three effects of complement system:
1. Enhances inflammatory response, e.g.: attracts phagocytes
2. Increases phagocytosis through opsonization or immune adherence
3. Creates Membrane Attack Complexes (MACs)  Cytolysis
 Classic Pathway Alternative pathway
Specific acquired immunity Non-specific innate immunity
Initiated by antibody Bacterial endotoxin, capsule
Interaction of all components C1, C4, C2 are by-passed
Properdin system not involved Properdin system is involved

Biological Function of complement:

1) Bacteriolysis, cytolysis
2) Function of complement fragments
a. Opsonisation: C3b, C4b
b. Mediator of inflammation: C3a, C4a, C5a
c. Chemotaxis: C3a, C5a, C567
3) C-dependent virolysis
4) Immunological regulation: C3, CR1, CR2, C3b
➢ Interferons (IFNs):
• are proteins (glycoproteins) released from viral infected cells.
• Host-cell-specific but not virus-specific
• 2 types: – Type I (IFN- and IFN-)
– Type II (IFN-)
• Cells producing IFNs – Mononuclear phagocytes (IFN- )
– Fibroblasts (IFN- )
– Macrophages and TH1 Cells (IFN- )
• Recombinant interferons have been produced. However short-acting and
many side-effects.
Modes of antiviral activity:
◆ Down regulate viral protein synthesis
◆ Activate NK-cell
◆ Up regulate MHC class I/II expression

➢ Cytokines:
o Interleukin – 1 (IL-1) induces fever and the production of acute phase proteins
o Some of which are antimicrobial and opsonise bacteria.
 Opsonisation
What Is Opsonisation?
The process by which bacteria or other cells become more readily and more
efficiently engulfed by phagocytes after attachment to opsonins.

Opsonins (Complement or Antibodies):

✓ Are proteins coat bacteria and promote attachment of
micro-organism to phagocyte … Opsonisation.
✓ The most common opsonins are IgG, IgA and C3b
•IgG: in blood
•IgA: in mucosal surfaces (airways, urogenital
system, gut)
•C3b: in the complement system
Why is Opsonisation important?
o Because pathogens and immune cells are negatively charged.
o So it is not easy for an immune cell to move close enough to a pathogen
o Opsonisation solves this issue since they have receptors which identify and
bind to protein molecules on immune cells.
Cells of innate immunity
Phagocytes PMNLs (Neutrophils)

Natural killer cells Mononuclear phagocytes

(monocyte-macrophage)
 Phagocytosis
What is a Phagocyte?
A cell that engulfs and digests material such as cell debris and microbes
Types of Phagocytes
 The professional phagocytes: (have receptors on their surfaces that can detect
harmful objects)
1 Neutrophils (Rapid response team, Short life span 1-2 days in the tissue,
For every neutrophil in the circulatory system, there are 100 more waiting
in the bone marrow)
2 Monocytes,
3 Macrophages (Live for weeks to months, One litre of human blood
contains about six billion phagocytes)
4 Dendritic cells,
5 Mast cells.
 Non-professional phagocytes: (phagocytosis is not
their principal function)
1 Fibroblasts, which can phagocytose collagen in
the process of remolding scars will also make
some attempt to ingest foreign particles
2 Osteoclast
 What is Phagocytosis?
The process of recognition, binding, ingestion and digestion of a pathogen by
exposing them to toxic chemicals

Steps of phagocytosis:
1) Chemotaxis
2) Recognition and attachment
3) Engulfment and creation of
phagosome
4) Fusion of phagosome with
lysosome
5) Destruction and digestion
6&7) Residual body → Exocytosis
 1) Chemotaxis:
• It is the directed movement of a cell
(Neutrophils) to the site in the tissues where
the concentration of chemotactic factors
(chemoattractant) is highest.
• The most potent chemotactic factors are the
C5a complement component.

2) Recognition and attachment

 Direct binding: Non-encapsulated microorganisms are easily phagocytosed
and killed within macrophages.
 Indirect binding: Encapsulated microorganisms require the production of
antibody in order to be effectively phagocytosed (Opsonisation). Once
engulfed, however, they are easily killed.

TLRs are a class of proteins that play a key role in the innate
Toll-like immune system by recognition of invading pathogens and
receptors (TLRs) activation of immune responses against them
✓ TLRs on Macrophage, Denderitic cells, epithelial cells
➢ The structures usually present on the surface of a bacterium, fungal cell, parasite,
or virus, are called pathogen-associated molecular patterns (PAMPs)
➢ The receptors that recognize PAMPs are called pattern recognition receptors
(PRRs)
➢ Toll like receptors (TLRs)
are the most important PRRs

Recognition of microbes and infected cells occurs via:

SIGNALS RECEPTORS
TLRs, RIGs, NOD,
Pathogen-Associated
Microbes Mannose receptor,
Molecular Patterns Phagocytes
Dectins, Scavenger
(PAMP)
receptors

Damage-Associated Damaged/ Antibodies, complement Mucosae and

Molecular Patterns Infected system, antimicrobial
(DAMP) circulating
cells peptides
3) Engulfment and creation of phagosome:
❖ Cytoskeleton of phagocyte rearranges to form arm like extensions (pseudopods)
that surround material being engulfed.
❖ Phagocytic cell engulfs invader, forming a membrane-bound vacuole called a
phagosome.
4) Fusion of phagosome with lysosome
❖The phagosome moves along the cytoskeleton to fuse with lysosomes (filled with
various digestive enzymes like lysozyme and proteases) and creates a
phagolysosome.
❖In neutrophils, membrane-bound bodies are granules.

5) Destruction and digestion

❖In present of Oxygen: sugars is metabolized by aerobic respiration producing
highly toxic oxygen products (superoxide, hydrogen peroxide, singlet oxygen,
hydroxyl radicals).
❖As available O2 is consumed metabolic pathway switches to fermentation
producing lactic acid and lowering pH.
1. Oxygen-dependent intracellular
When a phagocyte ingests bacteria, its oxygen consumption increases (respiratory
burst), produces reactive oxygen-containing molecules that are anti-microbial. The
oxygen compounds are toxic to both the invader and the cell itself.
2. Oxygen-independent intracellular: not effective as oxygen-dependent ones.
Four main types:
1) The uses of electrically charged proteins (Cationic proteins; cathepsin)
which damage the bacterium's membrane.
2) The uses of lysozymes; break down bacterial cell wall.
3) The uses of lactoferrins, which are present in neutrophil granules and remove
essential iron from bacteria.
4) The uses of proteases and hydrolytic enzymes; digest proteins of bacteria
Intracellular Killing

Oxygen Dependent Oxygen Independent

Reactive oxygen Reactive Nitrogen

species (ROS) species (RNS)
6) Exocytosis
Membrane-bound vesicle containing digested material fuses with the plasma
membrane. Material is expelled to the external environment.

Microbial Evasion of phagocytosis

Mechanism Examples
▪ Inhibit adherence: M protein, capsules S. pyogenes,
S. pneumoniae
▪ Kill phagocytes: Leukocidins S. aureus
▪ Lyse phagocytes: Membrane attack complex Listeria monocytogenes
▪ Escape phagosome Shigella
▪ Prevent phagosome-lysosome fusion HIV
▪ Survive in phagolysosome Coxiella burnetti
Inflammatory barriers
Tissue damage leads to inflammatory response
Purpose: ▪Destroy pathogen
▪Limit spread of infection
▪Pave way for tissue repair
4 cardinal signs:
❑ Acute-phase proteins (Chemical mediators) activated
❑ Complement proteins
❑ Cytokines
❑ Specialized proteins such as fibrinogen and bradykinin

The Three Stages of Inflammation:

1. Vasodilation and increased vessel permeability due to histamine (and
other cytokine) release  edema
2. Phagocyte migration and phagocytosis
❑ Margination and diapedesis (emigration)
❑ Chemotaxis
❑ Pus formation
3. Tissue repair and regeneration depends on type of tissue
Margination Diapedesis

Inflammatory process
 Fever: Abnormal High body temperature
▪ Hypothalamus acts as body’s thermostat. It releases prostaglandins that reset
the thermostat
▪ Endotoxin causes phagocytes to release IL–1. IL-1 is an endogenous pyrogen
▪ When no more IL–1, body temperature falls (crisis).
▪ > 40.7C (105F) can be dangerous (Tachycardia, acidosis, dehydration)
▪ Death at temp. > 44 - 46C

Beneficial effects of moderate fever:

▪ Inhibited pathogen growth
▪ Increased cellular metabolism  e.g.:
o Increased transferrin production
o Increased IL–1 activity  T cell production 
o Faster repair mechanisms
Constitutional factors that modify innate immunity
(1) Species
Resistance to a pathogen are shown by all the members of a species.
e.g. - All human beings are unsusceptible to plant pathogens.
- Mumps not seen in dogs or cats
(2) Racial
Different races within a species may show differences in susceptibility to
infections.
e.g. - Resistance to falciparum malaria in parts of Africa.
- Blacks more prone to diphtheria & influenza

(3) Individual
Differences in innate immunity among different individuals in a race.
e.g. Identical susceptibility & immunity to TB & leprosy seen in homozygous
twins, but not in heterozygous.
 Factors affecting innate immunity in an individual:
1) Age : low levels of immunity at the 2 extremes of life.
Fetus/ Neonates ……. immature immune system
Old age ….. deteriorated immune system, physical abnormalities.
2) Hormonal influences: Increased susceptibility in endocrine disorders like
a) Hormone deficiency in Diabetes, hypothyroidism, adrenal dysfunction ➔
reduced resistance
b) Elevated corticosteroids in pregnancy & stress ➔ reduce resistance
(Corticosteroids ➔ inhibit Ab formation & anti-inflammatory & anti-
phagocytic effect)
c) Prepubertal girls’ vagina susceptible to gonococci

3) Nutrition: Malnutrition results in reduced humoral & cell mediated immunity

Paradoxically,
a) Some infections may not manifest in malnourished individual e.g. malaria
b) Some viruses do not multiply in tissues of malnourished host.
c) Mantoux test negative in kwashiorkor