Innate Immunity

and
Complement System

Shashika Dayarathna
Department of Immunology and Molecular Medicine
09/05/2023
Learning objectives

• Describe how the innate immune system recognize pathogen.

• Describe the complement system and its function.

• Explain the mechanism of complement regulation and consequences of complement

dysregulation.

• Explain the acute phase proteins, their role in inflammation.

• Describe the innate immune responses to bacteria, virus and fungi.

Recall..

• What is immunity?

• Innate Vs Adaptive Immunity?

• Cells of Innate immunity?

Defensive barriers in Innate Immunity
•Anatomic
•Physiologic
•Phagocytic
•Inflammatory
❑ Defensive barriers in Innate immunity
Epithelial barriers prevent pathogen entry into the body’s interior
 Antimicrobial Proteins and Peptides kill would-be invaders
Examples?

Psoriasin

➢ Competition for body’s resources with normal flora ?

How do innate immune cells recognize microbes and damaged tissue?

Variety of surface receptors

Some directly recognize PAMPs (pathogen-associated molecular

patterns).

What are PAMPs?

Specific conserved molecular components of microbes
Usually present in many copies on the surfaces of microbes
Ex: cell wall of bacteria and fungi
can be expressed on non-pathogenic microbes
MAMPs (microbe associated molecular patterns)

Most PAMPs which induce phagocytosis are cell wall components…

Complex carbohydrates (Eg: Mannans, Β-glucans)
Lipopolysaccharides (LPS)
Peptidoglycans
Surface proteins

DAMPs ?
How do innate immune cells recognize microbes?

PAMPs are recognized by PRRs (pattern recognition receptors)

Cell associated: TLRs, CLRs, RLRs, NLRs

Soluble: Pentraxins, Collectins, Ficolins, Complements
 Inflammasomes

What are they?

Multiprotein enzymatic complexes

When/ how are they formed?

Form in cytosol in response to tissue infections
or cell injury/ stress

What do they do?

Generate biologically active forms of the
inflammatory cytokines IL-1b and IL-18

How do they produce those active cytokines?

A PRR complex
→ when activated it produces inflammatory cytokines
→ plays a role in protection against infection and pathogenesis of many diseases
Cells of Innate Immune system and their function
From previous lesson?

Phagocytes:
Specialized in phagocytic functions
Primarily Macrophages and DCs

DCs:
How do they promote different types of
adaptive immune responses?

ILCs:
ILC1, 2 and 3
Defense against viruses, helminths
Allergic inflammation, Intestinal barrier function, lymphoid organgenesis

NK cells:
Kill infected/ abnormal cells
Produce IFN-γ → activates macrophages to destroy phagocytosed microbes
 NK cell activity

Direct killing by distinguishing abnormalities

A reduction in the display of class 1 MHC molecules
and the unusual profile of surface antigens displayed
by some tumor/viral infected cells

ADCC – Antibody dependent cell mediated

cytotoxicity
NK cells express CD16 – a membrane receptor for the
Fc region of IgG
Some tumor cells/ cells infected by certain viruses
display antigens
Abs against them mediate NK cell toxicity against them

Chediak-Higashi syndrome-impairment in neutrophils, macrophages and NK cells- Increased incidence of lymphomas

Phagocytes and phagocytosis …?

• A key mechanism for eliminating pathogens

• The cellular uptake (eating) of particulate
materials such as bacteria
• Phagocytic cells make up the next line of
defense against pathogens who have
penetrated the epithelia.
 Opsonization
• Enhancement of phagocytosis
• If an antigen is coated with the appropriate antibody, the complex of Ag-Ab
binds to Ab receptors on the MØ membrane more readily than Ag alone

Opsonins
• Soluble phagocytosis-enhancing proteins (Soluble PRRs)
• Bind to conserved, repeating components on the
surfaces of microbes
• Once bound to microbial surfaces, they are recognized
by membrane opsonin receptors on phagocytosis
Examples:
MBL (mannose- binding lectin)- in blood & respiratory fluid
L- ficolin- in blood
SP-A and SP-D found in the blood, mucosal secretions in the lungs etc

• Individuals with MBL deficiencies;

Predisposed to severe respiratory tract infections, especially pneumococcal pneumonia
Protective against tuberculosis
Killing mechanisms for phagocytosed microbes

Binding of microbes to phagocytes via PRRs or opsonins and opsonin receptors

Activate signaling pathways

Trigger actin polymerization

Formation of membrane extensions around the microbe particle

Phagosome formation & internalization of microbe

Phagosome formation & internalization of microbe

Fusion of phagosome with lysosomes (in neutrophils, with preformed 10 & 20 granules)
 The resulting phagolysosome contains an arsenal of antimicrobial agents..!

Killing and degradation of internalized microbes

Oxygen independent Oxygen dependent

• Antimicrobial proteins and • Specialized molecules that
peptides (including defensins & mediate oxidative attack
cathelicidins)
• Low pH
• Acid-activated hydrolytic enzymes Occurs in;
(including lysozyme & proteases) Reactive oxygen species (ROS) - Neutrophils
Reactive nitrogen species (RNS) - Macrophages
Needs high oxygen concentration - dendritic cells
Chronic granulomatous disease – CGD Oxygen is provided by Respiratory burst

Defects in subunits of NADPH oxidase Cell’s O2 uptake increases in several fold.

Increased susceptibility to some fungal and bacterial infections
Macrophages are the major scavenger cells in body (identifying DAMPs)

“Eat me” signals- by altered membrane components of aged/damaged cells

“Don’t eat me” signals- by normal cells

• Healthy cells don’t express DAMPs

• Some tumor cells express healthy cell markers (ex:CD47) to evade phagocytosis
❑ Acute Inflammatory response Redness and
swelling with
heat,
pain and
loss of function
❑ Acute Inflammatory response
If the infection or tissue damage is not resolved
Vasodilation Can lead to chronic inflammatory state
Rise of the blood volume in the area Can cause more local tissue damage
Heats the tissue and causes tissue redness (erythema)
Potential systemic consequences

Increase in capillary permeability

Influx of fluid and cells
Exudate has a high protein content
Accumulation of exudate contributes to tissue swelling

Influx of phagocytes from the capillaries into tissues

Margination
Extravasation (Diapedesis)
Chemotaxis

Phagocytosis
Release lytic enzymes → can damage nearby healthy cells

Resolution
Clearance of invading pathogens, dead cells and damaged tissue
Activation of systemic acute phase response
Initiation of wound healing
Induction of adaptive immune responses
Acute phase proteins

• The proteins whose concentrations rise during the acute phase of inflammation
• Induced by signals that travel through the blood from sites of injury or infection.
• Major signals responsible: TNFs, IL-1, and IL-6

Eg:
Many complement proteins→ contribute to both innate and adaptive immune responses
MBL→ opsonin, initiates complement activation
CRP→ opsonin, activates complement mediated attack on pneumococcal bacteria
serum amyloid protein and PTX → opsonins, activators of complement pathway
SP-A, SP-D → opsonins, protection against lung infections
Fibrinogen → regulate the coagulation
The complement system …?

• Includes more than 30 soluble and cell-bound proteins

• Their biological activities affect both innate and acquired immunity

• It straddles innate and acquired immunity, contributing to each in a variety of ways.

• After initial activation, the various complement components interact, in a highly regulated cascade, to carry out several
basic functions
Complement Activation
The early steps to form C5b can occur by the;
➢ classical pathway
➢ alternative pathway
➢ lectin pathway.
The final steps that lead to a membrane attack are the same in all pathways.
Functions of Complement proteins
Why does the complement system need a tight regulation?

How?

• Includes highly labile components that undergo spontaneous inactivation if they are not stabilized by
reaction with other components.

• A series of regulatory proteins can inactivate various complement components

• C3b undergoes spontaneous hydrolysis by the time it has diffused 40 nm away from the C4b2a or C3bBb
convertase enzymes, so that it can no longer bind to its target site
Deficiencies of regulatory proteins

• Hereditary angio-oedema:
Deficiency of C1INH

• Paroxysmal nocturnal
haemoglobinuria:
Deficiencies in CD55 and CD59
Discussion
Innate immune responses

Bacteria Virus Fungi

? ? ?