THE IMMUNE SYSTEM - LECTURE 1

NORMAL IMMUNE SYSTEM

Protect from infectious pathogens
Innate: natural, or native, immunity: ready to react to infections even before they occur.
Specifically recognizes and combats microbes and damaged cells.
Adaptive: acquired, or specific, immunity: consists of mechanisms that are stimulated
by (adapt to) microbes and non-microbial substances. Capable of recognizing
microbial and non-microbial substances.

Components of Innate Immunity

Epithelial/endothelial barriers:
o Skin, gastrointestinal tract, and respiratory tract
o Provides mechanical barriers to the entry of microbes
o Produce antimicrobial molecules such as defensins
o Lymphocytes located in the epithelia combat microbes
Phagocytic cells:
o Monocytes and neutrophils circulate in blood and can rapidly be recruited to
any site of infection
o Macrophages (mature monocytes) are present in all tissues
Sense the presence of microbes and other offending agents
o Ingest (phagocytose) invaders and destroy them
Dendritic cells: in epithelia, lymphoid organs, and most tissues
o Capture protein antigens and display peptides for recognition by T
lymphocytes
o Stimulate the secretion of cytokines
Mediators that play critical roles in inflammation and anti-viral
defense
Natural killer (NK) cells
o Early protection against many viruses and intracellular bacteria
Other cells:
o Mast cells: produce many mediators of inflammation
o Innate lymphoid cells: look like lymphocytes but have features of innate
immunity
Plasma proteins
o Complement system - proteins that are activated by microbes using the
alternative and lectin pathways
In adaptive immunity it is activated by antibodies using the classical
pathway
o Mannose-binding lectin and C-reactive protein - coat microbes and promote
phagocytosis
o Lung surfactant - provides protection against inhaled microbes

Pattern Recognition Receptors

Recognize microbial components that are shared among related microbes
o Often essential for infectivity (thus cannot be mutated)
Damage-associated molecular patterns - recognize molecules released by injured and
necrotic cells
Located in all the cellular compartments where microbes may be present
o Plasma membrane receptors detect extracellular microbes
o Endosomal receptors detect ingested microbes
o Cytosolic receptors detect microbes in the cytoplasm

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Toll-Like Receptors (TLRs)
10 TLRs in mammals - each recognizes a different set of microbial molecules
Signal a common pathway that leads to the activation of transcription factors:
o NF-B - stimulates synthesis and secretion of cytokines and the expression of
adhesion molecules
Critical for the recruitment and activation of leukocytes
o Interferon regulatory factors (IRFs) - stimulates production antiviral cytokines
(type I interferons)
Germline loss-of-function mutations affecting TLRs and their signaling pathways
o Rare but serious immunodeficiency syndromes

NOD-Like Receptors (NLRs)

Cytosolic receptors
Recognize a wide variety of substances
o Products of necrotic cells (e.g., uric acid and released ATP)
o Ion disturbances (e.g., loss of K+)
o Some microbial products
Several NLRs signal via the inflammasome (a cytosolic multiprotein complex)
o Gain-of-function mutations in one of the NLRs result in a periodic fever
syndrome, called autoinflammatory syndrome
Can treat with IL-1 antagonists
The NLR-inflammasome pathway may also play a role in:
o Gout
o Obesity-associated type 2 diabetes
o Atherosclerosis
o Other

Other Receptors for Microbial Products

C-type lectin receptors: plasma membrane of macrophages and dendritic cells
o Detect fungal glycans and elicit inflammatory reactions
RIG-like receptors: cytosol of most cell types
o Detect nucleic acids of viruses and stimulate production of antiviral cytokines
G proteincoupled receptors: neutrophils, macrophages, and most leukocytes
o Recognize bacterial peptides containing N-formylmethionyl
Mannose receptors
o Recognize microbial sugars (mannose) phagocytosis of the microbes

Reactions of Innate Immunity

Inflammation
o Via cytokines, products of complement activation, and other mediators
Antiviral defense
o Via Type I interferons
Also provides stimulus for the adaptive immune response

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ADAPTIVE IMMUNITY

Adaptive Immunity
Develops later and is more powerful than innate immunity
Consists of lymphocytes and their products (antibodies)
Highly diverse receptors recognize a vast array of foreign
substances (antigens)
2 types:
Humoral immunity
Protects against extracellular microbes and their toxins
MediatedbyBlymphocytesandantibodies
Cell-mediated (or cellular) immunity
Defense against intracellular microbes
Mediated by T lymphocytes

Cells of the Immune System: B and T Lymphocytes

Immune surveillance
Constantly circulate via blood and lymphatics to lymphoid and other tissues
Lymphocytes:
Nave: not encountered the antigen for which they are specific
Once they are activated by recognition of antigens:
Effector cells: eliminate microbes
Memory cells: remember the antigen and can react rapidly and
strongly to it in future encounters

Lymphocyte Diversity
10^12 lymphocytes (1 trillion) in a healthy adult
Recognize 107 to 109 (10 million to 1 billion) different antigens
Lymphocytes can respond to multiple antigens, but once exposed to one
Undergoes clonal selection (all lymph with same specificity are clones)
Number lymphs specific for any one antigen is:
<1in100,000to1in1million
Generated by somatic recombination of the genes that encode the receptor proteins
During lymphocyte maturation (in the thymus for T cells and the bone
marrow for B cells)
Gene segments recombine in random sets and variations
Mediated by enzymes: RAG-1 and RAG-2 (recombination activating genes)
Inherited defects in RAG proteins result in failure to generate
mature lymphocytes
Recombined T and B cells
T-cell receptor (TCR)
Immunoglobulin (Ig) in B cells
Molecular assays can determine if a lymphocyte proliferation is polyclonal
(non-neoplastic) or monoclonal (neoplastic)

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T Lymphocytes
3 types:
o Helper T lymphocytes stimulate B lymphocytes to make antibodies and
activate other leukocytes (e.g., phagocytes) to destroy microbes
o Cytotoxic T lymphocytes (CTLs) kill infected cells
o Regulatory T lymphocytes limit immune responses and prevent reactions
against self antigens
Majority of lymphocytes in blood and tissue are T lymphocytes
Recognize specific cell-bound antigen via a specific TCR
o 95% of these are heterodimers made up of an and a polypeptide chain
TCR recognizes antigens presented by major histocompatibility complex (MHC)
molecules on the surfaces of APCs
o This MHC restriction ensures that T cells see only cell-associated antigens
Small population of mature T cells TCR
o Recognizes peptides, lipids, and small molecules, without assistance from
MHC proteins
o Aggregate at epithelial surfaces (e.g.skin, GIand urogenital tracts)
Small subset of NK-T cells express a very limited diversity of TCRs
o Recognize glycolipids that are displayed by the MHC-like molecule CD1
Other proteins that assist the TCR complex:
o CD4 and CD8 (mutually exclusive)
~60% of mature T cells are CD4+ and about 30% are CD8+
o CD4+T cells-cytokine-secreting helper cells that assist macrophages and B
lymphocytes
Bind to class II MHC molecules (coreceptor)
o CD8+T cells-cytotoxic (killer) Tlymphocytes (CTLs) that destroy host cells
harboring microbes
Bind to class I MHC molecules (coreceptor)

B Lymphocytes
~10-20% of circulating lymphocytes
Also present in:
Lymph nodes, spleen, and mucosa-associated lymphoid tissues
B cells recognize antigen via the B-cell antigen receptor complex
Membrane-bound antibodies of the IgM and IgD isotypes
Present on the surface of all mature nave B cells
After stimulation by antigen, they develop into plasma cell
Can secrete 100s-1000s of antibody molecules per second
Other molecules that are essential for signaling:
Type 2 complement receptor (CR2, or CD21)
Also used by Epstein-Barr virus (EBV) to enter and infect B cells
CD40 - receives signals from helper T cells

APCs

Dendritic Cells
Most important APC for initiating T-cell responses www.dendriticcellresearch.com
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 Have numerous fine cytoplasmic processes
Keys for antigen presentation:
o Located at the right place to capture antigens
Under epithelia (Langerhans cells in the skin)
Interstitia of all tissues where antigens may be produced
o Express many receptors for capturing and responding to microbes
TLRs and lectins
o Recruited to the T-cell zones of lymphoid organs in response to microbes
o Express high levels of MHC and other molecules needed for
presenting antigens to and activating T cells
Follicular dendritic cells: germinal centers of lymphoid follicles in the spleen and lymph
nodes
o Trap antigens bound to antibodies or complement proteins
o Present antigens to B cells and select the highest affinity B cells

Macrophages
Mononuclear phagocytes
Process antigens from phagocytosed microbes and proteins
o Present peptide fragments to T cells
T cells can activate macrophages and enhance their ability to kill ingested microbes
Phagocytose and destroy microbes that are opsonized (coated) by IgG or C3b

Natural Killer (NK) Cells

Function is to destroy irreversibly stressed and abnormal cells
o E.g. virus-infected cells and tumor cells
o Do so without prior exposure to or activation by these microbes or tumors
o Early line of defense
~5-10% of peripheral blood lymphocytes
Do not express TCRs or Ig
Surface molecules:
o CD16 an Fc receptor for IgG
Lyse IgG-coated target cells called antibody-dependent cell-
mediated cytotoxicity
o CD56 function not known
Regulated by a balance between activating and inhibitory receptors
o Next slide
Also secrete cytokines (IFN-)
o Activates macrophages to destroy ingested microbes
NK cells are regulated by many cytokines:
o IL-2 and IL-15 stimulate proliferation cells
o IL-12 activates killing and secretion of IFN-

Innate Lymphoid Cells (ILCs)

Recently identified (and still much to learn about them)
Populations of lymphocytes that lack TCRs but produce cytokines similar to T cells
o NK cells are first defined ILC
o Different subsets of ILCs produce IFN-, IL-5, IL-17, and IL-22
Functions of ILCs:
o Early defense against infections
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 o Recognize and eliminate stressed cells
o Provide cytokines that influence the differentiation of T lymphocytes

MHC Molecules
Key role in adaptive immune system
Function: display peptide fragments of protein antigens
Linked to many autoimmune diseases
Involved in rejection of transplanted organs
o Major Histocompatibility Complex
MHC molecules are called human leukocyte antigens (HLA)
Genes that encode HLA molecules are clustered on chromosome 6
o Highly polymorphic
o Thousands of alleles of MHC genes
o Individuals HLA alleles are pretty unique

MCH Class I Molecules

Expressed on all nucleated cells and platelets
HLA-A, HLA-B and HLA-C
Display peptides derived from proteins located in the cytoplasm
o E.g. viral and tumor antigens
Recognized by CD8+ T lymphocytes
o Class I MHC-restricted

MCH Class II Molecules

Mainly expressed on macrophages, B cells, and dendritic cells
Encoded by HLA-D region
o 3 subregions: HLA-DP, HLA-DQ, and HLA-DR
Display antigens that are internalized into vesicles
o Typically from extracellular microbes and soluble proteins
Recognized by CD4+ T cells
o Class II MHC-restricted

MCH Molecules
HLA Haplotype - Individuals inherit one set of HLA genes from each parent
o Typically two different molecules for every locus
o Polymorphism = almost innumerable combination
o Everyone is unique (except identical twins) makes organ transplantation
difficult
Key roles in regulating T cellmediated immune responses
o Determine what antigens a person reacts to
E.g. Ragweed pollen
Many autoimmune and other diseases are associated with particular HLA alleles

Cytokines: Messenger Molecules of the Immune System

Secreted proteins
Mediate many cellular interactions and functions of leukocytes
Can act on the cells that produce them
o Autocrine actions
Can act on neighboring cells
o Paracrine
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 Can act at a distance
o Endocrine

Cytokines contribute to different types of immune responses

Innate immune responses produced by macrophages, dendritic cells, NK cells and
others
o Induce inflammation and inhibit virus replication
o TNF, IL-1, IL-12, type I IFNs, IFN-, and chemokines
Adaptive immune responses produced mainly by CD4+ T cells
o Promote lymphocyte proliferation and differentiation and to activate effector
cells
o IL-2, IL-4, IL-5, IL-17, and IFN-
Some cytokines stimulate hematopoiesis
o Called colony-stimulating factors
o Role is to increase leukocyte numbers
o GM-CSF and IL-7

Cytokine and Therapy

Antagonists
o Block harmful effects of cytokines, inflammation, and tissue-damaging
responses
o E.g. - Rheumatoid arthritis
Dramatic responses to TNF antagonists
Administer cytokines to boost reactions
o E.g. - hematopoiesis and defense against some viruses

Cellular Tissue

Tissues of the Immune System

Generative Lymphoid Organs (primary/central)
o Thymus: T cells develop
o Bone marrow: Production of all blood cells and where B lymphocytes mature
Peripheral Lymphoid Organs (secondary)
o Lymph nodes
o Spleen
o Mucosal and cutaneous lymphoid tissues
o Tissues concentrate antigens, antigen-presenting cells, and lymphocytes
Optimizes interactions among these cells and the development of
adaptive immune responses

Lymph Nodes
Nodular aggregates of lymphoid tissues located along lymphatic channels throughout
the body
Lymph carrying antigens slowly suffuses through lymph nodes
o Antigen-presenting cells in the nodes are able to sample the antigens of
microbes
Dendritic cells pick up and transport antigens of microbes from epithelia and tissues via
lymphatic vessels to the lymph nodes

The Spleen
Abdominal organ
Serves the same role in immune responses to bloodborne antigen as lymph nodes do in
responses to lymph-borne antigens
Blood entering the spleen flows through a network of sinusoids
Antigens are trapped by dendritic cells and macrophages in the spleen

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Cutaneous and Mucosal Lymphoid Systems
Located under the epithelia of the skin and the gastrointestinal and respiratory tracts
o >1/2 the bodys lymphocytes are in the mucosal tissues
Many of these are memory cells
Respond to antigens that enter through breaches in the epithelium

Putting it all together

Antigen enters body Activation of innate immune system

Antigen recognition
Activation of lymphocytes to proliferate and
differentiate into effector and memory cells
Elimination of the antigen
Decline of the response
Memory cells survive

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HYPERSENSITIVITY REACTIONS
Sensitized
o Persons been previously exposed to an antigen
Hypersensitivity
o Excessive or harmful reaction to antigen
Can be exogenous environmental antigens (allergies) or endogenous self antigens
(autoimmune)
o E.g. Dust, pollens, foods, drugs, microbes, and various chemicals
Responses range from itching of the skin to bronchial asthma and anaphylaxis
Due to: imbalance between the effector mechanisms of immune responses and the
control mechanisms that limit responses
Associated with variety of susceptibility genes

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IMMEDIATE (TYPE I) HYPERSENSITIVITY

Allergies triggered by allergen (antigen)

o Ingested (e.g. peanut), inhaled (e.g. ragweed) or injected (e.g. bee sting)
Rapid immunologic reaction
Person who was previously sensitized
Antigen binds to IgE antibody on the surface of mast cells
May occur as a systemic disorder or as a local reaction
Reactions range from rash to fatal when severe

Immediate reaction: vasodilation, vascular leakage, and depending on the location, smooth
muscle spasm or glandular secretions
Minutes after exposure and subsides in a few hours

Late-phase reaction: (e.g., allergic rhinitis and bronchial asthma)

2-24 hours after exposure (without additional exposure) to antigen
Lasts for several days
Infiltration of tissues with eosinophils, neutrophils, basophils, monocytes, and CD4+ T
cells

Mast Cells
Throughout the body, especially near blood vessels, nerves and in subepithelial tissues
o Explains local immediate hypersensitivity reactions often occur at these sites
Have cytoplasmic membrane-bound granules
Activated by the cross-linking of high-affinity IgE Fc receptors (basophils too)
o IgE-coated mast cells are sensitized
I.e. sensitive to subsequent encounter with the specific antigen
Also triggered by:
o Complement components C5a and C3a (elicit reactions that mimic
anaphylaxis)
o Chemokines (e.g., IL-8)
o Drugs (e.g., codeine and morphin)
o Adenosine
o Melittin (present in bee venom)
o Physical stimuli (e.g., heat, cold, sunlight)

TH2 cells produce cytokines

IL-4: stimulates class switching of B cells to IgE and promotes the development of
additional TH2 cells
IL-5: development and activation of eosinophils
IL-13: enhances IgE production and acts on epithelial cells to stimulate mucus secretion

TH2 cells, mast cells and epithelial cells: produce chemokines that attract more TH2 cells and
other leukocytes

Preformed Mast Cell Mediators

Released first
Vasoactive amine: histamine

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 o Causes intense smooth muscle contraction, increased
vascular permeability, and increased mucus secretion
Enzymes: neutral proteases (chymase, tryptase) and several acid hydrolases
o Cause tissue damage
o Acts on precursor proteins to
Generate kinins
Activate components of complement (e.g.,C3a)
Proteoglycans
o Heparin: anticoagulant Chondroitin sulfate

Lipid Mast Cell Mediators

Reactions in the mast cell membranes lead to activation of phospholipase A2
o Converts membrane phospholipids to arachidonic acid Converted to
leukotrienes and prostaglandins
Leukotrienes C4 and D4 are the most potent vasoactive and spasmogenic agents
known
o 1000s times more active than histamine
Prostaglandin D2
o Causes intense bronchospasm and increased mucus secretion
Platelet-activating factor (PAF)
o Causes platelet aggregation, release of histamine, bronchospasm, increased
vascular permeability, and vasodilation

Mast Cell Cytokines

TNF, IL-1, and chemokines
o Promote leukocyte recruitment (typical of the late-phase reaction)
Inflammatory cells release additional waves of mediators (including
cytokines)
Cause epithelial cell damag
Epithelial cells can also produce chemokines
IL-4
o Amplifies the TH2 response

Late-Phase Reaction
Leukocytes (especially eosinophils) are recruited which amplify and sustain the
inflammatory response
o Without antigen
Eosinophils damage tissue via release of:
o Proteolytic enzymes
o Major basic protein
o Eosinophil cationic protein
This late-phase reaction is a major cause of symptoms in some type I hypersensitivity
disorders
o E.g., allergic asthma.
o Treatment requires broad-spectrum anti-inflammatory drugs
o Anti-histamine drugs are only useful for the immediate reaction

Allergies - Genetics
Atopy: increased propensity to develop immediate hypersensitivity reactions
o Have higher serum IgE levels and more IL-4 producing TH2 cells
Studies in patients with asthma show linkage to polymorphisms in several genes
o Some genes are located on chromosome 5
o Genes encoding the cytokines
o Linkage also noted to chromosome 6, close to the HLA complex

Allergies - Environment
Exposure to environmental pollutants is a predisposing factor
o Likely more important than genes
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 o Incidence of many allergic diseases is increasing in developed countries
Viral infections of the airways are triggers for bronchial asthma
Bacterial skin infections are strongly associated with atopic dermatitis

Allergies - Environment
Hygiene Hypothesis
o Decrease in infections during early life associated with higher incidence
o Exposure to microbial agents educates the immune system in early
childhood and even prenatally
o This hypothesis is difficult to prove
Microbiome

Non-atopic Allergy
~20-30% of immediate hypersensitivity reactions
Triggered by non-antigenic stimuli:
o Temperature extremes and exercise
Does not involve TH2 cells or IgE
Thought due to mast cells that are abnormally sensitive to activation

Systemic Anaphylaxis
Vascular shock, widespread edema, and difficulty in breathing
o Within minutes after exposure: itching, hives, and skin erythema appear
o Followed by a striking contraction of respiratory bronchioles and respiratory
distress
o Laryngeal edema hoarseness and further compromises breathing
o Vomiting, abdominal cramps, diarrhea, and laryngeal obstruction follow
o Patient may go into shock and even die within the hour
Occurs in sensitized individuals (but they may not know it yet)
Hospital:
o Foreign proteins (e.g., antisera), hormones, enzymes, polysaccharides, and
drugs (e.g., penicillin)
Community:
o Food allergens (e.g., peanuts, shellfish)
o Insect toxins (e.g., bee venom)
Extremely small doses of antigen may trigger anaphylaxis
o Even tiny amounts used in skin testing for various forms of allergies

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TYPE II HYPERSENSITIVITY

Antibody-Mediated (Type II) Hypersensitivity

Can be autoantibodies: antibodies specific for normal cell or tissue antigens
o or
Antibodies to exogenous antigens, such as chemical or microbial proteins

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Antibody-Mediated Cell Destruction and Phagocytosis in Disease
Transfusion reactions
o Cells from an incompatible donor react with and are opsonized by
preformed antibody in the host
Hemolytic disease of the newborn (erythroblastosis fetalis)
o Maternal IgG anti-erythrocyte antibodies cross the placenta and cause
destruction of fetal red cells
Autoimmune hemolytic anemia, agranulocytosis, and thrombocytopenia
o Antibodies to their own blood cells, which are then destroyed
Certain drug reactions
o Drug acts as a hapten by attaching to plasma membrane proteins of red
cells and antibodies are produced against the drug-protein complex
Responsible for tissue injury in some forms of
o Glomerulonephritis
o Vascular rejection in organ graft
o Others

Antibody-Mediated Cellular Dysfunction in Disease

Myasthenia gravis
o Antibodies reactive with acetylcholine receptors in the motor end plates of
skeletal muscles block neuromuscular transmission muscle weakness
Graves disease
o Antibodies against the thyroid-stimulating hormone receptor on thyroid
epithelial cells stimulate the cells hyperthyroidism

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TYPE III HYPERSENSITIVITY

Immune ComplexMediated (Type III) Hypersensitivity

Antigen-antibody complexes produce tissue damage mainly by eliciting inflammation
at the sites of deposition
o Typically in vessel walls
Antigens can be exogenous or endogenous
Immune complexmediated diseases preferentially involve:
o Kidney (glomerulonephritis)
o Joints (arthritis)
o Small blood vessels (vasculitis)

Immune Complex Disease Pathogenesis

Complement-fixing antibodies (i.e., IgG and IgM)
o Induce the pathologic lesions of immune complex disorders
Complement proteins can be detected at the site of injury
Consumption in active disease decreased serum levels of C3
o Can be used to monitor disease activity
Organs where blood is filtered at high pressure to form other fluids, like urine and
synovial fluid, tend to concentrated immune complexes

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TYPE IV HYPERSENSITIVITY

T CellMediated (Type IV) Hypersensitivity

Caused by inflammation resulting from cytokines produced by CD4+ T cells and cell
killing by CD8+ T cells

CD4+ T CellMediated Inflammation

Naive CD4+ T cells recognize peptides displayed by dendritic cells and secrete IL-2
stimulate proliferation of antigen-responsive T cells
Antigen-stimulated T cells differentiate to TH1 or TH17 cells based on the cytokines
produced by APCs
Repeat exposure to anWgen TH1 cells secrete cytokines (IFN-) activated
macrophages
o If activation is sustained, continued inflammation and tissue injury
Activated TH17 cells secrete IL-17, IL-22, chemokines, and other cytokines
o Recruit neutrophils and monocytes to the reaction Produce IL-21
amplifies the TH17 response

Clinical Examples of CD4+ T Cell Mediated Inflammatory Reactions

Classic example is the tuberculin reaction
o Intracutaneous injection of purified protein derivative (PPD, also called
tuberculin), a protein- containing antigen of the tubercle bacillus

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 o Previously sensitized individuals reddening and induration of the site
appear in 8 to 12 hours and peaks at 24 to 72 hours

Clinical Examples of CD4+ T Cell Mediated Inflammatory Reactions

Tuberculous infection
o Persistent tubercle bacilli
o Colonizes lungs or other tissues
o Infiltrate is dominated by macrophages after 2-3 weeks
o Sustained activation macrophages undergo a morphologic transformation
into epithelioid cells (large epithelium-like cells with abundant cytoplasm)
Granuloma: aggregate of the epithelioid macrophages, usually
surrounded by lymphocytes

Granulomatous inflammation
Associated with strong TH1-cell activation and high-level production of cytokines such
as IFN-

CD8+ T CellMediated Cytotoxicity

T cellmediated disease
o E.g. Type 1 diabetes
o Graft rejection after organ transplantation
Plays a role in reactions against viruses
o Some cases it is responsible for cell damage that accompanies the infection
(e.g., in viral hepatitis) Involved in killing tumor cells
o Tumor-associated antigens are presented on the cell surface

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