MICROIMM

2500A

Innate Immunity
Part II

Dr. Veronica Guariglia

Sep 14th 2022
Innate Immunity
IMMUNE SYSTEM

Innate Adaptive
Immunity Immunity

Barriers
T-cell B-cell
Cells
immunity immunity
Immune signaling
Innate and Adaptive Number of
pathogens

Immunity
BARRIERS

INNATE CELLS & PROTEINS

ADAPTIVE IMMUNITY

Janeway's Immunobiology, 10e

Innate Immunity
And
biological!

Part 1: Barriers to infection

Part 2: Cellular response
Part 3: Innate signaling
Learning Objectives
By the end of this lecture, you should be able to:

1) Know the cells that comprise the innate arm of the immune system

2) Explain how these cells (neutrophils, macrophages, immature dendritic cells, and
natural killer cells) provide innate immune defense
-where do they arise from?
-where do they reside?
-what mechanisms do they use to destroy a pathogen?

3) Describe the process of phagocytosis and apoptosis

4) Explain the mechanism by which innate immune cells recognize pathogens

Innate Immune Cells

Phagocytes Lymphocytes
Innate Immune Cells
Myeloid Origin:

Neutrophils
Monocytes/Macrophages
Dendritic Cells

Lymphoid Origin:

Natural Killer Cells

Dendritic Cells
NK cells
Key Innate Immune Cells
Myeloid Origin:
Destroy
Neutrophils extracellular pathogens
Monocytes/Macrophages by PHAGOCYTOSIS
Dendritic Cells

Lymphoid Origin:
Destroy
intracellular viruses
Natural Killer Cells
by APOPTOSIS
How are pathogens detected?
Recruitment and activation of immune cells requires pathogen detection

Phagocytes can exit the blood rapidly

& migrate to sites of infection
Pathogen Recognition and Detection
Recruitment and activation of immune cells requires pathogen detection

Induce
How do phagocytosis
cells
“sense”
the
presence of
pathogens?

Expand
response
Created with BioRender
Pattern Recognition Receptors (PRRs)
https://www.immunology.org/
• PRRs are expressed at the surface and within
many cell types (WBC, some lymphocytes, and
some epithelial cells)
• Recognize conserved, invariant regions of
pathogens
• Can also recognize damage or infection signals
• Specificity is genetically encoded
4 Families of PRRs:
• Toll-like receptors (TLR)
• C-type lectin receptors (CLR)
• Nucleotide oligomerisation receptors (NLR)
• RIG-1 like receptors (RLR)
Pattern Recognition Receptors (PRRs)

?
Pathogen Associated Molecular Patterns (PAMPs)

PAMPs include sugars, lipids, proteins, viral nucleic acids.

https://www.immunology.org/
Key properties of PAMPs

• Only expressed by pathogens (not self molecules)

• Highly conserved (low mutation rate). Critical for

microbial survival or pathogenesis

• Ideal for pathogen detection

• Detection of PAMPS is critical to the initiation of immune

responses
Examples of PAMPs
Examples of PAMPs
PAMPs critical for microbe’s structure PAMP critical for microbe’s pathogenicity
(integrity) (ability to infect)
Peptidoglycan & Lipotechoic Acid in cell wall of Bacterial Flagellin
Gram Pos bacteria
LPS in the cell wall of Gram Neg bacteria
Mannan in the cell wall of yeast
Glucan in the cell wall of fungi

Viral PAMPs
Viral nucleic acids
Capsid and surface proteins
Transmission electron microscopy
of hyperflagellated Salmonella
enterica serovar Typhimurium
Melchjorsen Viruses 2013, 5(2), 470-527 Tomoyasu et al. 2002. J Bact. 184:3.
PRRs bind PAMPs
- Each TLR has a distinct
repertoire of specificities.

- Usually PAMPs from

extracellular pathogens

- Surface TLRs recognize

components on the outside
of pathogens (e.g., LPS,
peptidoglycan, and flagellin)

- Endosomal TLRs recognize

components released during
degradation (e.g., bacterial
and viral nucleic acids)
 PRRs bind PAMPs PRRs and their corresponding Candida albicans PAMPs

PAMPs

Candida albicans is an opportunistic

surface
pathogenic yeast (fungus) that is a PRRs
common member of the human gut
microbiome.

phagocyte
intracellular
PRRs

https://www.cdc.gov/fungal/diseases/candidiasis

Mod from Infect. Immun. 80(4):1304, 2012

PRRs bind PAMPs
One type of PRR can bind to many different PAMPs (from different microbes)

This redundancy increases the repertoire of pathogens that can be detected

We only have a limited number of PRRs (< 100)

PAMPs

PRRs
PRRs bind PAMPs
Phagocytosis
PRRs bind to PAMPs Pseudopods engulf pathogen Phagosome formation

Illustration by Gary Kaiser https://bio.libretexts.org/Bookshelves/Microbiology/Book%3A_Microbiology_(Kaiser)

Key Innate Immune Cells
Myeloid Origin:
Destroy
Neutrophils extracellular pathogens
Monocytes/Macrophages by PHAGOCYTOSIS
Dendritic Cells

Lymphoid Origin:
Destroy
intracellular viruses
Natural Killer Cells
by APOPTOSIS
Phagocytosis
Cell “eating”
PRRs

PAMPs

Killing occurs within

the phagolysosome so
it does not damage the
phagocyte.
 Phagocytosis

Flannagan RS, Heinrichs DE. Front Microbiol. 2018 Dec 12;9:3084. doi:
10.3389/fmicb.2018.03084. PMID: 30619165; PMCID: PMC6299164.
The Phagolysosome
The phagosome is innocuous at first. Phagosome-lysosome fusion makes it highly
bacteriocidal

Most ingested
pathogens can be
killed in the Acid
phagolysosome. Enzymes
Antimicrobials
Created with BioRender
Antimicrobial Properties of the Phagolysosome
1) Low pH.
2) The NADPH oxidase -> reactive oxygen
1. pH 4-5
species. 4.

3) Myeloperoxidase (MPO) can transform 6.

3. Bleach
H2O2 into hypochlorous acid (Bleach). 2. ROS

4) Lactoferrin captures Fe2+ that is essential 5.

for bacterial growth

5) Defensins form pores.
6) Lysozyme degrades peptidoglycan.
Uribe-Queroland Rosales. Front Immunol. 2017 Oct 24;8:1368.
Key Innate Immune Cells
Myeloid Origin:
Destroy
Neutrophils extracellular pathogens
Monocytes/Macrophages by PHAGOCYTOSIS
Dendritic Cells
Neutrophils
- Majority (50% to 70%) of blood circulating leukocytes

- Exit blood and rapidly enter infected tissues

- Swarm in large numbers to the site of infection in

response to inflammatory molecules (main component
of pus and can be used as a measure of infection)

- Life span of only a few days

Neutrophils
Complete Blood Count
Neutrophils
Neutropenia
Abnormally low levels of neutrophils

Patients are highly susceptible to

deadly infections with a wide range of
pathogens and commensal
microorganisms
https://www.osmosis.org/learn/Neutropenia

Can restore neutrophil levels via blood transfusion (of neutrophil rich blood
fractions).

If untreated, innate immune response fails to control infection so pathogen can spread
rapidly, even into blood → Septic Shock (Neutropenic Sepsis)
Macrophages
- Professional phagocytes
- Remove pathogens and damaged host cells
- Differentiate from monocytes that circulate in the blood and
enter tissues
- Also have a role in adaptive immunity
- Resident macrophages of tissues have specialized functions
depending on location.
- Long life span (months to years!)
Macrophages
Tissue macrophages are strategically located throughout
the body and perform important immune surveillance
activities, including:
-phagocytosis
-antigen presentation
-immune suppression
Dendritic Cells
Immature dendritic cells circulate in
blood and reside in tissue below/among
epithelial cells

Mature dendritic cells have important

roles in adaptive immunity

Janeway's Immunobiology, 10e

Dendritic Cells
DC role & function depends on maturation state

Immature DC Mature DC

maturation

Created with BioRender

role in innate immunity initiates adaptive immunity

phagocyte antigen presentation
Key Innate Cells - Phagocytes
Key Innate Immune Cells
Myeloid Origin:
Destroy
Neutrophils extracellular pathogens
Monocytes/Macrophages by PHAGOCYTOSIS
Dendritic Cells

Lymphoid Origin:
Destroy
intracellular viruses
Natural Killer Cells
Natural Killer Cells

Cytotoxic, cell killers that circulate in blood

Kill intracellular viruses (virus infected cells and cancer cells)

NK cells have many cytoplasmic granules that contain

the cytotoxic enzymes perforin & granzyme
Natural Killer Cells Converged granules

Dispersed granules
NK Target

Cell
Perforin and death
granzyme

Mace et al. Immunol Cell Biol. 2014

NK cytotoxic granules migrate to cell-to-cell

contact area to release perforin and
granzyme and cause cell death in target cells.
Hang Yu Lin. https://sunnybrook.ca/
Natural Killer Cells
Virus infected cell

NK cell

Perforin forms a pore in the

cell membrane.

This allows granzyme to enter

the virus-infected cell.

Granzyme degrades nucleic Apoptosis: cell death

acids
and triggers apoptosis
Granzyme also degrades viral
nucleic acids to prevent viral
replication
Lopez et al. 2012. Trends in Immunology33:8
Natural Killer Cells

NK cells are an early component of the

response to viral infection.

After cytokine release (IFN, TNF, and IL-

12), a wave of NK cells follows.

Together they control viral replication.

Individuals with NK cell deficiency are

susceptible to frequent herpesvirus
infections.

Janeway's Immunobiology, 10e

Key Innate Cells - Summary
Extracellular microbes:
Macrophages
Neutrophils
Dendritic Cells
destroy microbes via
PHAGOCYTOCIS

Macrophages DCs Neutrophils

Intracellular viruses:
Natural Killer cells
target virus-infected cells
for APOPTOSIS

What about
intracellular bacteria?
Survival to phagocytosis
Intracellular bacteria can survive phagocytosis to live/replicate inside a phagocyte.

inhibit lysosome-phagosome fusion

→ bacteria live in phagosome
resistant to lysosomal acids/enzymes
→ bacteria live in phagolysosome
X
X Control of these bugs may require T cell help
an arm of the adaptive immune system

X e.g. Salmonella, Mycobacterium, Listeria monocytogenes, S. aureus

so called professional intracellular pathogens
Survival to phagocytosis
L. monocytogenes can survive in the
cytoplasm and spread cell to cell
InlA

InlA: internalization

LLO: Listeriolysin, degrades the ActA

phagolysosome
LLO
ActA: Actin assembly-inducing
protein

Mod from N Engl J Med 1996; 334:770-776

Summary
Effector Function of Innate Immunity
Apoptosis

Intracellular
viruses

PRRs

Initiation of adaptive response

Summary
Innate Cell Where does it Pathogen Killed Mechanism of pathogen
(Myeloid/lymphoid origin) reside? intra/extra? type? destruction

How do cells detect pathogens?

What are the steps of phagocytosis?

Summary
Summary

Which of these cells originate from a myeloid progenitor?

a) Monocytes/Macrophages
b) Neutrophils
c) Dendritic Cells
d) NK Cells
Summary

True or False?

NK cells can kill cells infected with intracellular bacteria

Summary

True or False?

Innate immunity involves receptors formed prior to

pathogen exposure
Next Lecture

1. Innate Immunity
Part 1: Barriers to infection
Part 2: Cellular response
Part 3: Innate signaling
2. Introduction to Adaptive Immunity
3. T cell Immunity
4. B cell Immunity