MICROIMMUNOLOGY 2500A REVIEW NOTES 1

PART I: IMMUNOLOGY
The immune system is a network designed to protect the body from
pathogens
Pathogen: any microorganism capable of causing disease
 Immune system needs to decide if something is actually a threat
 Proteins and cells help to sense and eradicate threats

The Chain of Infection

Infectious diseases are called transmissible or communicable diseases because they must be
passed on through the chain of infection:
 The reservoir must be broken in order to break the chain
 This is done via vector control, water treatment, antibiotics, etc.
 Good hygiene practices help to eliminate the portal of entry/exit

Pathogenic Spread
Endemic: expected level of an infectious disease that is continuously present in a population
within a geographic area
Epidemic: refers to an (often sudden) increase in the number of new cases of a disease above
what is normally expected in that population in that geographic area
Outbreak: low-scale epidemic
Pandemic: widespread epidemic

Hematopoiesis
Hematopoiesis: developmental by which white blood cells differentiate into either myeloid
progenitor cells or lymphoid progenitor cells
 Not exclusive to WBCs (all blood cells are derived from hematopoietic stem cells)
 Hematopoiesis is regulated by cytokines
 Hematopoiesis peaks in teens; so children and elderly are more prone to infection
Myeloid Progenitor Cells: stem cells which will differentiate into phagocytes
Lymphoid Progenitor Cells: stem cells which will differentiate into lymphocytes

Passive Immunity
There are 3 types of immunity: passive immunity, innate immunity and adaptive immunity
Passive Immunity: immunity acquired from another individual via the transfer of antibodies
 Provides immediate, short-term immunity (antibodies are rapidly degraded in body)
 Can by acquired by a donor OR artificially OR maternally
o Newborns are granted immunity thanks to maternal antibodies transferred via
the placenta
o Breast milk also prolongs effects of passive immunity
 Wanes in 10 months; child must rely on innate immunity until ~1 year old
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Innate Immunity
Innate immunity is the first line of defense against pathogens (only defense against
plants/invertebrates)
 Aims to destroy pathogen at an early stage
o Comprised of barrier, innate cells, and innate proteins
 Innate immunity is granted at birth, so it is not unique to the host
 If it doesn’t work, it will work alongside adaptive immunity to destroy a pathogen

Innate Immunity—Barriers
Barriers are measures that work to physically block the entry of pathogens entirely
 Barriers enter via open wounds, respiratory tract, GI tract and reproductive system
 Epithelial cells provide a physical barrier against pathogens
o Epithelial cells have tight junctions and are rapidly renewable
o Upon pathogen contact, epithelial cell sends a signal to produce defensins
 Class of antimicrobial peptides
 Defensins attack ALL pathogens by forming a pore in the cell membrane
 The pore allows an influx of extracellular fluid into the microbe
 Epithelial cells undergo desquamation (shedding) in order to remove stubborn threats
o ALL cells (internal and external) undergo desquamation

Another type of barrier is mucus

 Epithelial cells called goblet cells secrete mucin
 Mucin mixes with water to make mucus
 Mucus traps pathogens in thick substance before they can infect cells in the host

Other barriers can be classified in three categories:

1. Mechanical Barriers: work by “flushing out” the pathogen
e.g. sneezing, coughing, blinking, peristalsis (vomit, diarrhea)

2. Chemical Barriers: work through the use of chemicals in the body

e.g. enzymes, lysozymes, low pH, defensins

3. Microbiological Barriers: work to destroy pathogens through the use of microbiota

 The microbiome is acquired from the mother from the vagina during birth
 Microbiota are associated with epithelial cells that line all pathogen entry points
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Innate Immunity—Innate Cells

Innate cells are split up into phagocytes (P), which phagocytose extracellular pathogens, and
lymphocytes (L), which destroy intracellular pathogens by apoptosing the host cell:

Neutrophils are the most abundant leukocyte (WBC)

 Undergo rapid hematopoiesis from myeloid progenitor cells
Neutrophils (P)  Exit blood and enter infected tissue quickly
 High abundance = short lifespan
Neutropenia: low neutrophil count (>1500/uL), resulting in increased
threat of bacterial (extracellular) infection
Macrophages arise from blood monocytes that have entered tissues
Macrophages (P)  Most reside just below epithelial cells; do not leave tissues
 Also play a role in adaptive immunity
Immature DCs circulate in the blood and the tissue below epithelial cells
Dendritic Cells  Immature DCs are phagocytes, while mature DCs play a role in
(P) adaptive immunity
NK cells are located in blood and kill intracellular threats very early
 NK cells bind to host cell, with their binding area becoming
Natural Killer polarized for effective release of toxic enzymes
Cells (L)  Perforin forms a pore in the host’s cell membrane
 Granzyme degrades the DNA/RNA of the host AND pathogen
 This induces apoptosis of the host, ending viral replication

The first responders to a pathogen WILL be IDCs and macrophages because they are
strategically located beneath the epithelial tissue.

Pattern Recognition Receptors (PRRs)

Phagocytes bind pathogens during phagocytosis using PRRs
 There are fewer than 100 PRRs, but the same PRR can bind multiple pathogens
 PRRs are expressed inside and on the surface of cells, in 4 classes:
o Toll-Like Receptors (TLR)  Nucleotide Oligomerisation Receptors (NLR)
o C-type Lectin Receptors (CLR)  RIG-1 Like Receptors (RLR)

Pathogen-Associated Molecular Patterns (PAMPs)

PRRs bind to molecules exclusively expressed by microbes called PAMPs like lock-and-key
 PAMPs may be sugars, proteins, lipids and nucleic acids
 PAMPs often play a critical role in a pathogen’s structural integrity and
pathogenicity
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Lysosomal Enzymes
Defensins Work by damaging the pathogenic cell membrane
Lysozymes
Works by producing toxic oxygen metabolites (H2O2 and HOCl)
NADPH Oxidase  Chronic Granulomodus Disease: victims are unable to
produce NADPH oxidase (quite fatal)
Nitric Oxide Synthetase Works by producing toxic nitric oxide

Phagocytosis (extracellular pathogen disposal)

A phagocyte’s PRR will recognize a PAMP on a pathogen, inducing cell-pathogen binding
 Phagocyte wraps around and internalizes pathogen, becoming a phagosome
 Phagosome fuses with lysosome to become a phagolysosome
 Lysosomal contents (acids and enzymes) are released into the phagolysosome
 Pathogen destruction occurs within the phagolysosome, so the phagocyte is unharmed
o One phagocyte can phagocytose several pathogens at once

o Phagocytosis can also by triggered by CR1 binding C3b (see next section)
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Innate Immunity—Innate Proteins

Cytokines are small proteins that serve to communicate with other cells and themselves
 Every activated nucleated cell secretes multiple cytokines at once
 In charge of cell regulation of hematopoiesis
 Involved in innate AND adaptive immunity

How Cytokines Work

During phagocytosis, PRR-PAMP binding induces cytokine production
 Produced cytokines (pro-inflammatory cytokines and chemokines) activate endothelial
cells
o Endothelial cells vasodilate; result is increased blood flow and increased blood
vessel permeability
o Immune cells are bound loosely to blood vessel wall, looking for point of entry
into infected tissue
o Vasodilation allows for a greater number of immune cells AND proteins to enter
into the infected tissue
 Leukocytes will NOT bind to or leave the arteries because arterial pressure is too high
for them to bind

Chemokines
Chemokines are chemoattractant cytokines: they attract cytokines by polarizing them
-important because immune cells are only needed in certain areas of the body
-released from the site of a wound, by the phagocytes disposing the pathogen

Inflammation and Pro-Inflammatory Cytokines

Pro-inflammatory cytokines are responsible for inducing vasodilation and increasing vascular
permeability
 Pro-inflammatory cytokines include TNF, IL-1 and IL-6

During inflammation, activated phagocytes release chemokines and pro-inflammatory

cytokines
 The effects of the pro-inflammatory cytokines are swelling, redness and heat
 The cytokines induce the migration of inflammatory cells into infected tissue
o Inflammatory cells release inflammatory mediators that cause pain

Fever
Fever is an early response to infection
 Useful because it destroys pathogens that cannot reproduce in elevated temperatures
 Sweat induced by fever contains defensins and antibodies (IgA)
 Activated phagocytes release pro-inflammatory cytokines and chemokines
o The cytokines enter the circulation and activate the hypothalamus
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o The hypothalamus subsequently produces prostaglandins, which cause fever

Acute Phase Proteins (APPs)

The body’s immediate response to a pathogen is called the acute phase response (APR)
 During the APR, cytokines travel to the liver, triggering hepatocytes to secrete APPs
 APPs travel to the site of infection via inflammation
 Complement Protein 3 (C3) is an APP that spontaneously cleaves into C3a+C3b
o C3b engulfs the pathogen, then binds to C3b receptors (CR1) expressed on
phagocytes, triggering phagocytosis
o C3a maintains inflammation by activating endothelial cells and attracting
phagocytes (acting like cytokines/chemokines)

Type I Interferons (IFNs)

Type I IFNs (IFN- IFN-ß) are antivirals secreted by cells infected by intracellular viruses
 PRRs inside infected cells bind viral nucleic acid (which acts as a PAMP)
 A cell signaling event follows, producing chemokines and type I interferons
o The chemokines attract NK cells
o Type I interferon activates NK
 Type I interferon induce the production of antiviral enzymes in healthy cells
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Adaptive Immunity
Adaptive immunity ensures an effective response against re-exposure to a particular pathogen
 It begins with a primary immune response, which serves to develop antibodies
 Subsequent exposures (memory immune response) are faster and larger scale
o Adaptive immunity works in tandem with innate immunity
 Primary lymphocytes involved are B cells (extracellular threats) and T cells (intracellular
threats)

Primary Lymphoid Tissues

B cells and T cells develop from primary lymphoid tissues (bone marrow and thymus)
 B cells: hematopoietic stem cell  lymphoid progenitor cell  sent to bloodstream
 T cells: hematopoietic stem cell  lymphoid progenitor cell  thymus
o In thymus, T cells express CD-4 (to become helper T cell) or CD-8 (to become
cytotoxic T cell)

Secondary Lymphoid Tissues

Naïve B cells and T cells respond to pathogens and become effector cells in these tissues.
 Where adaptive immunity is initiated
If peripheral tissue is infected, plasma containing pathogens enters
capillaries (becoming interstitial fluid)
Lymphatic System  Fluid enters lymphatic vessels to become lymph, which is
filtered by lymph nodes
 T cell- and B cell-specific zones in lymph nodes filter lymph
Spleen B cells and T cells in the spleen filter blood
 Destroy blood-borne pathogens
Mucosa-Associated MALT encompasses any tissues with a mucous membrane
Lymphoid Tissue  Respiratory, gastrointestinal and reproductive tracts
(MALT)  Swollen tonsils = sign of first time pathogenic infection

Mechanism of B Cell & T Cell Binding

B/T cells express surface receptors (BCR and TCR, respectively)
 BCR has 2 identical heavy and light chains bound to the cell via transmembrane region
 TCR has ONE - & ONE -chain, bound to the cell via transmembrane region
o Receptors are made up of a constant and a variable region (BCR has 2 VRs)
o Variable region binds antigen at the epitope
 Antigens are not specific to pathogens!
 Each B/T cell has specificity for ONE unique antigenic epitope
 B/T cells express many copies of a BCR/TCR with its own unique variable region
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o Specificity arises from random rearrangement of V(D)J genes, which alter the
shape of the variable region
 Once bound, naïve B/T cells undergo clonal expansion to become effector/memory
cells

Antigens and Blood Type

Antigens are the basis for Blood Transfusion Compatibility
 Blood type is based on expression of A, B and Rh (D) antigens on RBCs
 O-neg blood is the universal donor because it expresses no antigens
 AB+ blood is the universal acceptor because it expresses every type of antigen

T Cells & T Cell Binding

T cells are responsible for disposing of intracellular pathogens
 TCR is only capable of binding protein antigens
 Protein antigens = peptides
 TCR must be presented the peptide on a Major Histocompatibility Complex (MHC)
o MHC molecules are glycoproteins that bind hydrophobic epitopes (peptides)

MHC Class I Molecules

All nucleated cells have MHC class I molecules
 MHC class I molecules present molecules generated in the cytoplasm of the host cell
o Pathogens replicating in cytosol express proteins
o Proteasomes cleave proteins into endogenous peptides
o Endogenous peptides enter ER, where MHC class I molecules are found
o MHC I:peptide complex travels through Golgi complex to cell surface
 The complex is ultimately bound by a cytotoxic T cell (CTL)

MHC Class II Molecules

MHC class II molecules are only presented by “professional” antigen presenting cells (APCs)
 B cells, macrophages and mature DCs
 Macrophages and MDCs internalize pathogens via phagocytosis (B cells  endocytosis)
 Phagolysosome or endolysosome cleave pathogenic proteins into exogenous peptides
 Exogenous peptides bind to MHC class II molecules, and complex is presented on cell
surface

Immature Dendritic Cell Maturation During T Cell Response

DC involvement in immune response depends on the location of the DC in question
 SLT Resident DCs: IDCs meet pathogens in lymph, spleen and MALT
o DC become infected, presenting peptide on MHC class I, or
o DC phagocytose pathogens, presenting them on MHC class II
o DC are bound to stromal network, and T cells circulate in search of them
 Peripheral Tissue DCs: IDCs that phagocytose extracellular pathogens leave tissue and
migrate toward the lymph nodes via lymphatics
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o During migration, IDC matures, ultimately becoming an APC

NOTE: Only dendritic cells can trigger naïve T cells to produce effector T cells.

Effector T Cells—Cytotoxic T Cells (CTLs)

CTLs work identically to NK cells, with the exception that they must bind an MHC Class I
molecule in order to function
 Some viruses have evolved to block peptide presentation of peptides on the MHC I
o This may be by blocking proteasome digestion or the formation of the MHC-
peptide complex
o Only NK cells can kill these pathogens
 Cross-Presentation: process by which DCs present both endogenous and exogenous
peptides from the phagocytosis of extracellular pathogens

Effector T Cells—Helper T Cells (Th Cells)

Effector Th cells secrete cytokines
 Th1 cells are enhance the destruction of intracellular pathogens replicating in the
phagosome/phagolysosome of macrophages
o Typically prone to infections from these pathogens because they replicate in
vesicle-like vehicles
o Th1 cells bound to MHC class I: peptide complex release IFN-gamma
 Increases production of Lysosomal enzymes/Lysosomal fusion
o Th1 cells also create granulomas to “suffocate” pathogens
 Granulomas involve the creation of a hypoxic environment

Endocytosis (B Cell Pathogen Ingestion)

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