Cell biology and Immunology

The immune system (Chapter 24)

Lecture 5: Immune response pathways

P. Heeringa
Pathologie en Medische Biologie
UMCG
1
p.heeringa@umcg.nl
E-learning: Acute and chronic inflammation
(see link to the module on nestor)
 Infection
Pathogens can enter our body in different ways

routes of infection:

- broken skin barrier

cuttings, stings, burn-, bite-wounds

- inhalation, ingestion

Barriers are broken, pathogens enter our body

Sets off several interrelated reactions aimed to neutralize or kill the

invader
• What happens when extracellular bacteria
invade our body?

• What happens when viruses invade our body

(for the second time)?
Fig. 24.12 Immune responses to extracellular bacteria

Bacterial infections cause inflammation and

trigger specific immune responses.

Bacteria enter
extracellular fluid
from outside

© 2013 Pearson Education, Inc.

 Bacterial infections cause inflammation and
trigger specific immune responses. Bacteria enter
extracellular fluid
from outside
External
environment

Skin or mucous
membrane

ECF

Bacteria

Triggering of acute inflammation (‘ontvlamming”)

à protective reaction of vascularized tissue to
injury à Characterized by vascular changes
and involves soluble and cellular components of
the innate immune system

Capillary

© 2013 Pearson Education, Inc.

 The inflammatory response
Inflammation can be triggered by any form of tissue damage e.g. infection,
trauma, foreign body reactions, physical damage (burns, freezing), reduced tissue
oxygen (e.g. myocardial infarction) etc.

Inflammation involves vascular changes and cells and soluble factor of the
(innate) immune response

Goal:
- Damage control
- Neutralize or eliminate the intruder
- e.g. kill bacteria
- e.g. clear foreign body material
- Initiate tissue repair

- Inflammation = beneficial = protective

reaction involving the innate immune
system
 Types of inflammation

• Acute inflammation (max. few days)

Max. few days

Complete resolution
Tissue repair

• Chronic inflammation
– Long lasting (months, years)
– Caused by persistence of the inflammatory trigger
• (e.g. autoimmune disease)
– Consequence: tissue damage, loss of tissue function (fibrosis)
Acute inflammation is characterized by
5 clinical symptoms.

What do you see?

What do you feel?
The clinical signs of acute inflammation

The 5 (clinical) signs of inflammation

(as first described by Celsus)
• Heat (Calor)
• Redness (Rubor)
• Swelling (Tumor)
• Pain (Dolor)
• Loss of function (functio laesa)
(added by Virchow)
Acute inflammation involves cells and soluble
mediators of the innate immune system

Inflammation is strongly linked to activation of the

innate immune system
• Tissue resident cells (macrophages, dendritic cells, mast cells)
e.g in the skin

• Complement system (coagulation system, kinin system)- self

enhancing enzyme cascades

• Infiltrating immune cells (neutrophils, monocytes)

Acute inflammation is triggered by a whole
range of inflammatory mediators

• Tissue resident cells (macrophages, dendritic cells,

mast cells)
– Histamine, cytokines (e.g. IL-1, IL-6, TNF)

• Complement system, (coagulation system, kinin

system)- self enhancing enzyme cascades
– Chemotaxins (C3a, C5a), bradykinin

• Infiltrating immune cells (neutrophils, monocytes)

– Cytokines (e.g IL-1, IL-6, TNF)
 General characteristics of inflammatory
mediators

• Regulators of the inflammatory response

• Exert their function by binding to receptors on targets

cells

• Are pleiotropic and are redundant

– One mediator can target multiple effector cells
– One cell can be targeted by multiple mediators

• Short half-life (immediately active, fast resolution)

 Acute inflammation has four phases

• Acute inflammation has 4 phases

– Hyperemie (heat, redness)
• increased blood supply but reduced blood flow
– Exudation (swelling, pain, loss of function)
• leakage of plasma fluid (including plasma proteins) into the
tissue
– Infiltration
• Migration of leukocytes from the circulation into the tissue
– Resolution and Repair
• Termination of inflammation and initiation of tissue repair
(with or without scar formation)
 Phase 1 and 2: vascular changes

Hyperemia and exudation

are initiated by activated resident tissue e.g. infection
cells e.g. tissue macrophages (e.g. kupffer
cells in the liver), dendritic cells and
mast cells e.g.Toll-like receptor binding

• Heat (Calor)
Cytokine secretion
– vasodilation
• Redness (Rubor)
– vasodilation
• Swelling (Tumor) bradykinin
NO histamine
– Oedema
bloodstream
• Pain (Dolor)
– Oedema and chemical transudate
vasodilation
mediator (kinins) exsudate
• Loss of function (functio laesa) Increased permeability
oedema
 Bacteria enter
extracellular fluid
from outside
External
environment

Skin or mucous
membrane

ECF lyses 3. Lysis of microbes

Bacteria
(membrane attack
Membrane complex)
attack complex
activate

Increased permeability causes Complement

make
activate
plasma proteins (including proteins
Mast cells

complement proteins) to leak into

are
secrete

the tissue. Bacterial cell wall Chemotaxins Histamine

components activate the complement

system
increases
permeability
1. Inflammation: recruitment
and activation of neutrophils
(C3a en C5a)

2. Activate mast cells

(secretion of mediators
that attract neutrophils
cause vasodilation and
Capillary attract
Circulating leukocytes
increased vessel
permeability)
© 2013 Pearson Education, Inc.
Phase 3: Infiltration of leukocytes

e.g. infection

Decreased blood flow allows enhanced

interaction and adhesion of
leukocytes to endothelium

Cytokine secretion activates

endothelium àupregulation of
adhesion molecules
bradykinin
NO histamine
bloodstream

vasodilation transudate
exsudate

oedema
 Phase 3: Infiltration of leukocytes

1:Rolling

2:Adhesion

3:Transmigration
Proinflammatory cytokines
e.g. IL-1β, TNFα

18
Systemic effects of cytokines produced during
inflammation
When produced in high amounts, pro-inflammatory cytokines enter the bloodstream
and induce systemic effects

Increased mobility of C-reactive protein

Leukocytes (opsonin, activates
Enhanced phagocytosis complement)
Inhibit bacterial growth Complement factors
 innate immune cells catch pathogens at the border (phagocytosis) à
first defense
à migrate via the lymphatics to draining lymphnodes
à present the pathogens to the adaptive immune cells
Dendritic cells in the skin
(Langerhans’ cells) extracellular
Bacteria enter
fluid
External from outside
environment

Skin or mucous
membrane

ECF lyses
coat Bacteria
Membrane
attack complex
Opsonins activate

ingest and make

act as
Complement activate
disable
proteins
Mast cells
are
secrete
Acute phase
Phagocytes proteins Chemotaxins Histamine

increases
permeability

Antibodies

act as

Plasma proteins

Capillary Circulating leukocytes attract

© 2013 Pearson Education, Inc.

adaptive immune cells (T and B cells) become activated
migrate via the blood to the site of infection
cooperate in the elimination of the pathogen

Dendritic cells in the skin

(Langerhans’ cells) Bacteria enter
extracellular fluid
External from outside
environment

Skin or mucous
membrane

ECF lyses
coat Bacteria
Membrane
attack complex
Opsonins activate

ingest and present make

act as
Complement activate
disable antigens to
proteins
Mast cells
are
secrete
Acute phase
Phagocytes T H cells
proteins Chemotaxins Histamine
activate
increases
B lymphocytes permeability
become

secrete Antibodies

Plasma cells act as

Plasma proteins

Capillary Circulating leukocytes attract

© 2013 Pearson Education, Inc.

Adaptive immunity and innate immunity work
together: Phagocytosis

Complement
(innate)

+
Antibodies
(adaptive)

=
Enhanced phagocytosis

22
Adaptive immunity stimulates innate immune
function: T cells

Naive CD4+T cell

APC

23
 Phase 4: resolution and initiation of tissue
repair
When the pathogen is eliminated…

Inflammation declines spontaneously

Mediators of inflammation are produced in rapid bursts
only while the stimulus persists
Mediators have short half-lives and are degraded after
their release.
Neutrophils also have short half-lives in tissues and die by
apoptosis within a few hours after leaving the blood

Active termination mechanisms are induced

mechanisms including:
Release of anti-inflammatory cytokines, including
transforming growth factor-β (TGF-β) and IL-10, from
macrophages

Production of anti-inflammatory lipid mediators

 Acute inflammation: short summary

• Immediate, early response on tissue injury

Vasodilation (reduced blood flow)
Increased Vascular permeabilityà Leads to leakage of complement
factors into tissue and complement system activation

• Emigration of leukocytes into tissues

Recognition and removal of microbes and dead tissues
Macrophage activationà link to adaptive immunity!

• Resolution and repair

Involves passive (apoptosis of leukocytes) and active mechanisms
(anti-inflammatory mediators)
 Outcome 1: Complete resolution
Complete resolution- no scar formation
In cases with limited tissue damage

• Bloodflow normalized

• Inflammatory cells are gone

• Complete tissue repair

• Most ideal outcome

 Outcome 2: Scar formation/fibrosis

Resolution but with scar formation: In cases with extensive

tissue damage

Normal Injured
• E.g massive bacterial
infectionà extensive tissue
cell death due to toxic
products released by massive
amounts of neutrophils
à replaced by fibrous tissue
(i.e. increased deposition of
extracellular matrix proteins
e.g. collagen)

• E.g. heart attack à extensive Blue staining

tissue cell death à à Fibrous
replacement by fibrous tissue tissue
Outcome 3: Acute inflammation progresses to
chronic inflammation
When the initial trigger can not be cleared completely
or persists (e.g. autoimmune reactions)à
extensive fibrosis and organ dysfunction Chronic kidney disease

Autoimmune hepatitis, Alcoholic liver disease

Lung fibrosis

Organ dysfunction!
29
 Cell biology and Immunology
The immune system (Chapter 24)

Lecture 6: Immune response pathways

(virus)

30
What happens when a virus infects our
body (for the second time) ?
 Fig. 24.13 Immune responses to viruses

This figure assumes prior exposure to

the virus and preexisting antibodies.

Virus
invades
host

© 2013 Pearson Education, Inc.

 Life cycle of a virus
Viral propagation depends on host cell machinery
Semlike Forest virus “budding”

© 2013 Pearson Education, Inc.

 Pre-existing antibodies prevent infection

Assumes prior exposure to

the virus and preexisting antibodies.

- Virus antigen specific antibodies are present

and prevent infection
Preexisting antibodies
- Illustrates memory of the adaptive immune Virus
invades
systemà long-lived memory B cells host

t
lls ec
ce t inf
o
nn
ca
Uninfected host
cell

© 2013 Pearson Education, Inc.

Induction of Humoral immunity is the basis of
immunity induced by many vaccines: Examples
Infectious Vaccine Mechanism of
disease protective
immunity
Polio Oral attenuated polio Neutralization of virus
virus by mucosal IgA
antibody
Tetanus, diphteria Toxoids Neutralization of toxin
by systemic IgG
antibody
Hepatitis Recombinant viral Neutralization of toxin
envelope proteins by systemic IgG
antibody

Pneumococal Conjugate vaccines Opsonization and

Pneumonia composed of bacterial phagocytosis by IgM
Haemophilus capsular polysaccharide and IgG antibodies
and protein directly or secondary
to complement
activation
 Pre-existing antibodies prevent infection
but are not always adequate

Assumes prior exposure to

the virus and preexisting antibodies.

- Virus antigen specific antibodies prevent Preexisting antibodies

infection Virus
invades
host

t
lls ec
- Illustrates memory of the adaptive immune

ce t inf
o
nn
systemà long-lived memory B cells

ca
- Once the virus has infected a host cell,
Uninfected host
antibodies are no longer effective cell
MHC-I

- Other immune mechanisms must be Viral

antigen
activated Infected host cell

© 2013 Pearson Education, Inc.

 Macrophages ingest antibody coated virus
particles: Stimulate inflammation
Preexisting antibodies
Virus
invades
host Macrophage
ingests virus.

t
lls ec
ce t inf
MHC-II

o
nn
ca
Viral antigen

Macrophage presents
antigen fragments.
Uninfected host MHC-II
cell secretes
MHC-I Activates helper T cell
Interferon-a
Viral activates Cytokines
antigen antiviral
response.
Infected host cell
Inflammatory Helper T cell
response

Antibody coated virus particles are phagocytosed by macrophages

Macrophages get activated (e.g via endosomal PRRs) and secrete cytokines
à Stimulates the inflammatory response à more leukocytes are mobilised

© 2013 Pearson Education, Inc.

 Interaction of microbe with macrophage
stimulates inflammation

Pathogen

Endocytosis receptors(in this case Fc receptors)

TLR

Elimination of “danger”
Cytokines
activation macrophages (NO, ROI)

38
Important cytokines produced by macrophages
upon activation

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Importance of interferons in protection against
viral infections
Interferons α and β can be produced by activated macrophages but also
by the virus-infected host cells

ds viral RNA (PAMP) can be recognized by TLRs and other PRRs and lead to cytokine production

© 2013 Pearson Education, Inc.

 Macrophages ingest antibody coated virus
particles: Stimulate adaptive immunity
Preexisting antibodies
Virus
invades
host Macrophage
ingests virus.

t
lls ec
ce t inf
MHC-II

o
nn
ca
Viral antigen

Macrophage presents
antigen fragments.
Uninfected host MHC-II
cell secretes
MHC-I Activates helper T cell
Interferon-a
Viral activates Cytokines
antigen antiviral
response.
Infected host cell
Inflammatory Helper T cell
response

Antibody coated virus particles are phagocytosed by macrophages

The macrophages present viral peptides in the context of MHCII on their membrane
and activate antigen specific CD4+ helper T cells.

© 2013 Pearson Education, Inc.

 Interaction of microbe with macrophages
stimulates adaptive immunity

Pathogen

Endocytosis receptors (in this case Fc receptors)

TLR
CD4+ T cell

Co-stimulatory molecules, MHC class II Observation of “danger”

Cytokines

42
CD4 T cells are activated by peptides presented
by MHC class II molecules
CD4+ T lymphocyte

MHC class II

APC’s
“Professional” antigen presenting
cells that have phagocytosed and
processed (extracellular )pathogens

43
Antigen presenting cells and their function in
the immune response

44
 T cell-mediated responses: co-stimulation

Signal 1
Antigen
recognition

Co-stimulation Signal 2
 Preexisting antibodies
Virus
invades
host Macrophage
ingests virus.

t
lls ec
ce t inf
MHC-II

o
nn
ca
Viral antigen

Macrophage presents
antigen fragments.
Uninfected host MHC-II
cell secretes
MHC-I Activates helper T cell
Interferon-a
Viral activates Cytokines
antigen antiviral
response.
Infected host cell
Inflammatory Helper T cell
response
activates

Antigen specific CD4+ T

helper cells bind to viral Virus

peptides presented in
MHC II by macrophages B lymphocytes
and stimulate innate and become
adaptive immunity via cell-
cell interaction and
cytokine secretion.
Plasma cells
secrete

Antibodies

© 2013 Pearson Education, Inc.

 CD4+ T lymphocyte activation:
effector cells that orchestrate immune responses

Th-cells: different sub-sets….

different cytokine signatures….
different effector functions

Naive CD4+T cell

APC

47
 Preexisting antibodies
Virus
invades
host Macrophage
ingests virus.

t
lls ec
ce t inf
MHC-II

o
nn
ca
Viral antigen

Macrophage presents
antigen fragments.
Uninfected host MHC-II
cell secretes
MHC-I Activates helper T cell
Interferon-a
Viral activates Cytokines
antigen antiviral
response.
Infected host cell
Inflammatory Helper T cell
response
activates

Antigen specific CD4+ T

helper cells bind to viral Virus

peptides presented in
MHC II by macrophages B lymphocytes
and stimulate innate and become
adaptive immunity via
cell-cell interaction and
cytokine secretion.
Plasma cells
secrete

Antibodies

© 2013 Pearson Education, Inc.

 Illustrates: Specificity and memory of the adaptive
immune response

Antigen X and Y induce

a different immune response
(specificity)

The secondary response to

antigen X is more rapid,
larger and more effective
than the primary response
(memory)

© 2013 Pearson Education, Inc.

 Preexisting antibodies
Virus
invades
host Macrophage
ingests virus.

t
lls ec
ce t inf
MHC-II

o
nn
ca
Viral antigen

Macrophage presents
antigen fragments.
Uninfected host MHC-II
cell secretes
MHC-I Activates helper T cell
Interferon-a
Viral activates Cytokines
antigen antiviral
response.
Infected host cell
Inflammatory Helper T cell
response
activates
activates

CD4+ T helper cells also Attacked by cytotoxic T cells Virus

activate CD8+ cytotoxic T

cells T-cell
receptor B lymphocytes
Cytotoxic T cell become
Perforins,
granzymes

Plasma cells
secrete

Antibodies
Infected cell undergoes
apoptosis and dies.
© 2013 Pearson Education, Inc.
CD4+ T lymphocytes helps in CD8+ T cell
differentiation

IFNγ

51
 Preexisting antibodies
Virus
invades
host Macrophage
ingests virus.

t
lls ec
ce t inf
MHC-II

o
nn
ca
Viral antigen

Macrophage presents
antigen fragments.
Uninfected host MHC-II
cell secretes
MHC-I Activates helper T cell
Interferon-a
Viral activates Cytokines
antigen antiviral
response.
Infected host cell
Inflammatory Helper T cell
response
activates
activates

Antigen specific CD8+ Attacked by cytotoxic T cells Virus

cytotoxic T cells bind to

viral peptides presented in T-cell
receptor B lymphocytes
MHC I on infected cells Cytotoxic T cell become
Perforins,
and kill these cells. granzymes

Plasma cells
secrete

Antibodies
Infected cell undergoes
apoptosis and dies.
© 2013 Pearson Education, Inc.
 Recognition Activation and effect

Target cell killing

CTL

© 2013 Pearson Education, Inc.

 A functional immune response needs

Cooperation between all immune components

(innate and adaptive, humoral and cellular) in time.

the individual immune components have their own

basal activity

but…

their activity is enhanced to effective immunity by

interaction !
Same immunological principles are needed for effective
vaccines e.g against EBOLA and SarsCov-2
Same immunological principles are needed for a effective
vaccines e.g against EBOLA and SarsCov-2
An effective vaccine against EBOLA!
 Can you answer these questions?
and explain the underlying immunological mechanisms?

How is the immune system able to recognize pathogens that differ in

structure, life form and, size with a high level of sensitivity and
specificity?

Why do you only get some infections like chicken pox (waterpokken)
aka varicella zoster only once?

Why do subsequent immune responses to a pathogen occur more

rapidly and more effectively than previous immune responses?

How does the immune system provide a high degree of sensitivity and
specificity to the broad array of pathogens without attacking self?

58
 Next lecture

• Disorders when our immune system

overreacts or incorrectly responds to self
• Immuno-therapies