Chapter 21

Nonspecific Body Defenses and Immunity

G.R. Pitts, J.R. Schiller, and James F. Thompson, Ph

1. Innate (nonspecific) defenses

External body membranes Inflammation

Defense Systems

Antimicrobial proteins, phagocytes and other cells

2. Adaptive (specific) defenses

T cells and B cells

Innate Defense System

Surface Barriers
First line of defense: mechanical and chemical protection 1. Skin 2. Mucosal Membranes

Internal Nonspecific Defenses

1. 2. 3. 4. 5. Second line of defenses Phagocytes Natural Killer cells (NK lymphocytes) Inflammation Antimicrobial proteins Fever

Skin and Mucosal Membranes

Mechanical Protection
Epidermis
nose hairs, nails

Mucous membranes - line certain organ systems

mucus prevents drying, traps foreign things respiratory tract cilia sweep mucus out

Lacrimal apparatus -- tear glands and ducts

wash the eye to dilute microbial growth

Saliva - dilute microbes on the oral cavity Urine - flow dilutes, and acid pH helps kill, microorganisms Defecation and vomiting - expel toxins and microbes

Skin and Mucosal Membranes

Chemical Protection: reduce bacterial growth
Skin
sebum (unsaturated FAs) forms oily layer perspiration has fatty acids, salts (NaCl), and mildly acid pH

Lysozyme
in perspiration, tears, saliva, nasal secretions, other tissue fluids enzyme breaks down bacterial cell walls

Hyaluronic acid
gel-like matrix in most connective tissues slows the spread of many infectious agents

Gastric juice - stomach nearly sterile due to acid pH, ~2 Vaginal secretions mildly acid pH

Innate Defense: Phagocytes

Macrophages (derived from monocytes) are the chief tissue phagocytic cells
Free macrophages wander through tissues in search of microbes and cellular debris Fixed macrophages: Kupffer cells (liver), microglia (brain), dust cells (lungs)

Neutrophils become phagocytic when encountering infectious material Eosinophils are weakly phagocytic, deploy destructive granules against parasitic worms

Mechanism of Phagocytosis
Chemotaxis Adherence recognition of external carbohydrates and proteins
Aided by opsonins

Ingestion Killing and digestion

Innate Defense: Natural Killer Cells

Distinct group of large granular lymphocytes (NK lymphocytes = Null Killer lymphocytes) Nonspecific killers respond to the lack of selfantigens and to the presence of certain surface oligosaccharides Kill virus-infected body cells and some tumor cells by releasing various defensive molecules not by phagocytosis Act before the antigen-specific immune system is activated Secrete potent chemical signals that enhance the inflammatory response

Innate Defense: Inflammation

1. Inflammation Signs:
1. 2. 3. 4. 5. Redness Heat Swelling Pain Loss of Function

Function:
1. Prevent spread of damage 2. Dispose of pathogens and debris 3. Set stage for tissue repair

Stage 1: Vasodilation and increased vessel permeability

Inflammation

Macrophages and cells lining the gastrointestinal and respiratory tracts carry Toll-Like Receptors (TLRs) that recognize specific classes of microbes TLReceptor activation causes cytokine release
promotes inflammation & chemotaxis

Mast cells secrete histamine Other cells secrete various regulatory factors
Histamine, kinins, prostaglandins, leukotrienes, complement
http://www.komabiotech.co.kr/technical/review/toll_like_receptor.gif

Cause local vasodilation Increase capillary permeability resulting in edema

Inflammation: Stage 1
Edema increased plasma filtrate seeps into tissue spaces bringing some immune proteins
Helps to dilute harmful substances Increases supply of oxygen and nutrients needed for metabolism, inflammation and repair Allows entry of clotting proteins, which reduces the spread of mibrobes

Inflammation
Stage 2. Phagocyte moblization 1. Leukocytosis-inducing factors: increase neutrophil production 2. Margination (pavementing) 3. Diapedesis (amoeboid movement) 4. Chemotaxis of WBCs
neutrophils rapid arrival monocytes slower arrival

Inflammation
Stage 3. Tissue repair
Tissue regrowth and repair of damage or scar formation Pus
dead phagocytes and other WBCs, damaged tissue, and perhaps microbes if too numerous for effective removal by phagocytes, an abscess may develop

Effects of Inflammation
Increased blood flow results in increased local temperature and local cellular metabolism Increased capillary permeability and phagocytic migration to the injured tissue

Innate Defense: Antimicrobial Proteins

1. Attack microorganisms directly 2. Interfere with microbial reproduction The most important are:
1. Interferons 2. The Complement System 3. Transferrins which bind Fe2+ in plasma, inhibiting bacterial growth

Interferons (IFNs)
Produced by most tissue cells when infected by a virus Diffuses to uninfected cells and binds to surface receptors
stimulates macrophages and natural killer lymphocytes stimulates production of antiviral proteins which block viral replication inhibits growth of virally infected cells suppresses growth of tumor cells

Alpha IFN is used against:

hepatitis C virus herpes virus (genital warts)

The Complement System

20 plasma and cell membrane proteins that exist as inactive precursors When activated, the complement system functions to complement or enhance certain immune, inflammatory, and allergic responses Kills bacteria and certain other microbial cell types (our cells normally are protected from complement attack) Stimulates chemotaxis in leuckocytes Enhances the effectiveness of both nonspecific and specific defenses

Complement Pathways
Classical Pathway is triggered by the specific immune system
Requires binding of antibodies to antigens of invading organisms Complement C1 then binds to the antigen-antibody complexes (complement fixation)

Alternative Pathway is triggered by nonspecific interaction among factors B, D, and P, and microbial cell wall polysaccharides (complement fixation) Both pathways involve an enzyme cascade

Complement Pathways
Both pathways converge on C3, which cleaves into C3a and C3b C3b initiates formation of a membrane attack complex (MAC) MAC causes cell lysis by creating many hundreds of microscopic holes in the cells plasmalemma C3b is also an opsonin

Innate Defense: Fever

Pyrogens reset the temperature set-point in the hypothalamus Inhibits some microbes from growing Increases bodys metabolic rate, which speeds up immune defenses and tissue repair Increases effects of antimicrobial substances produced by the immune system Stimulates liver and spleen to sequester iron and zinc (needed by microorganisms)

High fevers are dangerous

Innate Defense System: Review

Surface Barriers
1. Skin 2. Mucosal membranes

Internal Nonspecific Defenses

1. 2. 3. 4. 5. Phagocytes Natural Killer cells (NK lymphocytes) Inflammation Antimicrobial proteins Fever

Adaptive Defense
The adaptive immune system:
Acts to immobilize, neutralize, or destroy foreign substances and cells Amplifies the inflammatory response and activates complement Is antigen-specific*, systemic, and has memory
*Recognizes specific foreign molecules

Has two interdependent arms

Humoral, or antibody-mediated immunity (AMI) Cellular, or cell-mediated immunity (CMI)

Adaptive Defense
Definitions:
Immunity: the ability of the body to defend itself against specific foreign invaders (molecules or cells) Immunogenicity: the ability to stimulate proliferation of specific lymphocytes and specific antibody production Reactivity: the ability of activated lymphocytes and their products, antibodies, etc., to interact with specific antigens

Adaptive Defense
Definitions:
Specificity: the antigen triggers focused immune defenses (from particular lymphocytes lineages) that respond only to the antigens of this foreign substance/cell Memory: the immune system produces clones of specific memory lymphocytes (T & B) which react rapidly when the particular foreign substance/cell is encountered again Specificity and memory differentiate this system from the nonspecific (innate) defenses

Adaptive Defense
Antigen any substance which provokes specific immune responses Antigenic determinants
Parts of antigens that trigger the specific immune response An antigen may be an entire microorganism or only small structures or subregions of large molecules

Most antigens are complex and express multiple types of antigenic determinants.

Chemical Nature of Antigens

Complete Ag: complex macromolecules - usually proteins (nucleo-, lipo-, glyco-) -sometimes carbohydrates or lipids
Are immunogenic & reactive

Incomplete Ag: smaller molecules (haptens)

react with antibodies but cannot cause an immune response without aid (protein carrier)
e.g., poison ivy, drug allergies

Adaptive Defense
Antigen receptor diversity
>1 billion different antigenic determinants are recognized by the body Genetic recombination shuffles and reorganizes different Ab genes

Major histocompatibility complex antigens (MHC)

unique to each individuals cells; help in identifying what is self versus foreign 2 classes of MHC antigens (markers)
class I MHC found on all body cells except RBC's class II MHC - only on antigen presenting cells (APCs), thymus cells, and activated T cells

Antigen-Presenting Cells (APCs)

APCs phagocytize, process, and present antigens to lymphocytes APCs do not respond to specific antigens APCs contribute to coordinating specific immunity
Macrophages Dendritic (Langerhans) cells B lymphocytes

The major initiators of adaptive immunity are APCs, which actively migrate to the lymph nodes and secondary lymphoid organs and present antigens to T and B cells

Class I MHC Proteins

Found on all cells, except RBCs Recognized by T lymphocytes and APCs Display peptides from endogenous antigens
Endogenous antigens are:
Associated with body cells Degraded by proteases and enter the endoplasmic reticulum Transported through special membrane channels Bound with MHC class I molecules on the ER membrane Migrate to the cell membrane as a complex: Ag -- MHC class I molecule

MHC Class I Proteins

This is a form of Antigen Presentation

Cancer cells often do something quite similar to the virus-infected cells. (Foreign MHC Class I Ags are the source of tissue transplant rejections.)

MHC Class II Proteins

1. Immune cell identity markers found only on mature B cells, some T cell classes, and antigen-presenting cells 2. MHC Class II proteins are synthesized in the ER 3. A phagosome containing a pathogen (with exogenous antigens) merges with a lysosome 1. MHC Class II proteins migrate into the phagosome where the antigen macromolecules are degraded and particular antigen peptides are bound to the MHC Class II markers

2. Ag-- MHC class II complex then migrates to the cell membrane and displays antigenic peptide for

MHC Class II Proteins

This is a key function of our APCs in most Ag-specific defenses.

Lymphocytes Provide Ag Specificity

B and T lymphocytes develop in bone marrow Lymphocytes mature and develop immunocompetence (ability to recognize specific antigen) in different locations
B cells mature in the bone marrow and provide Abmediated immunity T cells mature in the thymus and provide cell-mediated immunity

Immunocompetent B or T cells
Naive cells display a unique surface receptor for a specific antigen once mature
Receptor expression occurs before a cell encounters the foreign antigen it may later attack It is genes, not antigens, that determine which foreign substances our immune system will recognize and resist

Naive cells circulate to secondary lymphoid tissue where they may encounter antigens later
B and T cells become fully functional only after binding with their recognized antigen

Immunocompetent T Cells
T cells mature in the thymus under positive and negative selection pressures
Positive selection outer thymic cortex
Selects functional T cells which become both immunocompetent and potentially self-tolerant Non-selected cells die via apoptosis
Survive

Apoptosis

Apoptosis

Negative selection inner thymic cortex

Kill or regulate off T cells that react with self-antigens

Immunocompetent B Cells
B cells become immunocompetent and selftolerant in bone marrow
Some self-reactive B cells are killed by apoptosis (clonal deletion) Some self-reactive B cells can modify their anti-self properties (receptor editing)

Some self-reactive B cells are released from the bone and are inactivated by negative regulation (anergy)

Cell-Mediated Immunity
CMI is involved in most aspects of specific immune defense Three populations of T lymphocytes regulate specific immunity
Helper TH cells which carry CD4+ markers Suppressor TS cells Memory T cells

cytotoxic TC cells which carry CD8+ markers destroy tumor cells and virus-infected cells; they also attack transplanted cells and tissues

Cell -Mediated Immunity

Basic steps
1. Recognition by T lymphocytes of antigen presented by an antigen-presenting cell with matching MHC Class II markers 1. Proliferation and differentiation of T cells once activated 1. Production of clones of identical effector T cells capable of recognizing a specific antigen 1. Appropriate action (help, attack, memory, suppression) from T cell subclones

T Cell ActivationStep 1: Antigen Binding and Antigen Presentation

T Cell Activation- Step 2: Co-Stimulation

T cells must bind to MHC Class II surface receptors on an APC After co-stimulation with cytokines, T cells enlarge, proliferate, and form clones Activated T cells differentiate and perform functions according to their T cell class

T Lymphocyte Activity
Primary T cell response usually peaks within a week T cells then undergo apoptosis within a month Reduced activity parallels elimination of antigen This is a negative feedback control A few Memory T cells remain to respond to any future exposure to the same antigen

 Regulatory cells that play a central management role in the immune response

Helper TH Lymphocytes

Once primed by APC antigen presentation, TH cells:

Stimulate proliferation of other T cell classes Stimulate B cells that have already become bound to antigen

There is NO coordinated immune response without TH cell function

Helper TH Lymphocytes
TH cells interact directly with B cells that have antigen fragments on their surfaces bound to MHC Class II receptors TH cells express CD4+ cell identity markers TH cells stimulate B cells to divide more rapidly and begin antibody formation B cells may be activated without TH cell help by binding to T cellindependent antigens (certain microbial polysaccharides) Most antigens, however, require TH co-stimulation to activate B cells Cytokines released by T amplify nonspecific defenses

Cytotoxic Tc Lymphocytes
TC cells express CD8+ cell identity markers TC cells, or killer T cells, are the only T cells that can directly attack and kill other cells They circulate throughout the body in search of body cells that display the antigen to which they have been sensitized Their targets include:
Virus-infected cells Cells with intracellular bacteria or parasites Cancer cells Foreign cells from blood transfusions (WBCs and platelets) or

Cytotoxic Tc Lymphocytes
Bind to self/anti-self complexes on any body cell Infected or abnormal cells can be destroyed as long as appropriate antigen and co-stimulatory regulators (e.g., IL-2) are present [In contrast, Natural Killer cells activate their killing machinery when they bind to a different MHC-related cell surface marker on cancer cells, virus-infected cells, and transplanted cells]

 Secrete perforins which cause cell lysis by creating transmembrane pores Secrete lymphotoxin which fragments the target cells DNA Secrete gamma interferon which stimulates macrophage attack

Cytotoxic Tc Lymphocyte Actions

Suppressor Ts Lymphocytes
TS cells immune regulatory cells which release cytokines that suppress the activity of both T cells and B cells Generated when other specific T cell clones are generated Negative feedback control to bring the body back to normal after the battle has been won

Antibody-Mediated Immunity
Antigen challenge the first encounter between an antigen and a naive B lymphocyte Antigen presentation usually occurs in the spleen or a lymph node, but can occur in any lymphoid tissue Antigen presentation usually made by a macrophage, but some B cells can react directly against certain bacterial antigens Binding of the antigen to the B cells specific Ag receptor activates the B cell

Primary Response
Activated B cells grow and divide, forming clones bearing the same antigen-specific receptors and secreting the same antigen-specific Ab Most clone cells become plasma cells that secrete specific antibodies Clones that do not become plasma cells become B memory cells that can respond to subsequent exposures to the same antigen

Primary Response
Initial B cell differentiation, proliferation, and Ab synthesis requires time after the first Ag exposure Lag period: 3 to 6 days after antigen challenge Peak plasma levels of antibody are achieved in ~10 days Antibody molecules also reach the interstitial fluids, especially where inflammation exists Antibody levels then decline gradually if there is no additional Ag exposure

Secondary Response
Any subsequent exposure to the same antigen Sensitized memory cells (B and T) respond within hours Antibody levels peak in 2 to 3 days at higher plasma levels than in the primary response Activated B subclones generate antibodies that bind with greater affinity Plasma antibody levels can remain high for weeks to months

Primary and Secondary Antibody Responses

Immunological Memory
Immunization is possible because memory B cells and memory T cells persist after the initial Ag exposure with any subsequent exposure, the immune system responds more quickly, forcefully secondary response - antibodies produced during subsequent exposures are produced in greater quantities and have a greater attraction for antigen

Antibodies
Are unique soluble proteins secreted by activated B cells and plasma cells in response to an antigen Are capable of binding specifically with that antigen Constitute much of the gamma globulin fraction of plasma proteins Also called immunoglobulins

Basic Antibody Structure

Four polypeptide chains linked together with disulfide bonds The four chains bound together form an antibody monomer Each chain has a variable (V) region at one end and a constant (C) region at the other Variable regions of the heavy and light chains combine to form the antigen-binding site

Ag

Antibody Structure
Antibodies responding to different antigens have different V regions but the C region is the same for all antibodies in a given antibody class C regions form the stem of the Y-shaped antibody monomer and determine:
the class of the antibody the cells and chemicals to which the antibody can bind how an antibody class functions in eliminating antigens

Classes of Antibodies
IgD: monomer attached to the surface of B cells, important in B cell activation IgM: pentamer released by plasma cells during the primary immune response IgG: monomer that is the most abundant and diverse antibody in primary and secondary responses; crosses the placenta and confers passive immunity IgA: dimer that helps prevent attachment of pathogens to mucosal surfaces IgE: monomer that binds to mast cells and basophils, causing histamine release when activated

Antibody Functions
All antibodies form an antigen-antibody (immune) complex Antibodies do not directly destroy antigen, though they may immobilize or inactivate Ag Antibodies act as opsonins and tag Ag for immune attack and destruction Defensive mechanisms triggered by antibodies include neutralization, agglutination, precipitation, opsonization, and complement fixation

Antibody Mechanisms of Action

1. Neutralization: Antibodies bind to and block specific sites on viruses or exotoxins, thus preventing these antigens from binding to receptors on tissue cells Antibodies bind to the same determinant on more than one antigen forming antigen-antibody complexes that are cross-linked into large lattices 2. Agglutination: Cellular antigens are cross-linked, causing cell clumping 2. Precipitation: Soluble molecules are cross-linked into large insoluble complexes

Antibody Mechanisms of Action

4. Opsonization: Bound Abs facilitate phagocyte adherence 5. Complement Fixation: IgM and IgG antibodies bound to cellular Ags bind complement via the Classical Pathway
The complement cascade causes chemotaxis, opsonization, phagocytosis and cell lysis Complement activation enhances the inflammatory response

Summary of Antibody Actions

Figure 21.13

Monoclonal Antibodies
Monoclonal antibodies are purified tissue culture preparations of a specific antibody for a single antigenic determinant which are produced from descendents of a single B cell Commercially prepared monoclonal antibodies are used:
To provide passive immunity In research applications In clinical laboratory testing In the treatment of certain cancers

Adaptive Immunity: Summary

A defensive system with two interdependent arms (CMI & AMI) that uses lymphocytes, APCs, and specific molecules to recognize and destroy foreign substances Adaptive immune responses depend on the ability of its cells to:
Distinguish foreign from self molecules React with foreign substances (antigens) by binding to them Communicate with one another to effect a coordinated protective response specific to those antigens

Adaptive Immunity: Summary

To start an immune response, APCs, B and T lymphocytes must recognize foreign antigen Antigen-Presenting Cells and some B cells recognize and immediately bind to certain antigens in the blood, the extracellular fluid (ECF), or other tissue spaces More often, B and T cells only recognize antigen (protein fragments) when Ag is presented by the macrophages in combination with MHC Class II surface markers and stimulation is provided by Th lymphocytes

Summary of the Immune Response

Clinical Classification of Immunity

Active Immunity: the bodys own B and T lymphocytes encounter antigens and produce specific responses against them; immunological memory does occur
Naturally Acquired response to a microbial or parasitic infection Artificially Acquired response to a vaccine of dead or attenuated (weakened) pathogens

Passive Immunity: An outside source of immune cells or molecules is provided to a recipient; immunological memory does not occur; protection ends when the donated materials are naturally eliminated from the body
Naturally Acquired the mother to her baby via the placenta (IgG) or via lactation (colostrum/milk) (IgM & IgA) Artificially Acquired the injection of serum, gamma globulin, or leukocyte transfusion

Clinical Classification of Immunity

Organ and Tissue Transplants

The four major types of grafts are:
Autograft graft transplanted from one site on the body to another in the same person Isograft graft between identical twins (or clones); individuals with the same genotype Allograft graft between individuals that are not identical twins, but belong to same species Xenograft grafts taken from another animal species

Prevention of Graft Rejection

Donors are selected to minimize differences in MHC Class I antigens = HLA (human leukocyte antigens)
Unnecessary for routine blood transfusions since RBCs lack HLAs

Prevention of rejection is accomplished by using various immunosuppressive drugs

Survival and longevity of grafts have varying success

Immunosuppressive drugs depress the patients immune system so it is less effective in defending against pathogens and cancer

Pathologies: Immunodeficiencies
Human Immunodeficiency Virus
HIV enters certain cell types by receptor mediated endocytosis
infects primarily helper T cells attaches to the CD4 protein on cell surface

A retrovirus
carries its genetic material as RNA inserts its genetic material into host cell DNA with the enzyme reverse transcriptase cell makes copies of the virus, releases them for further infection

May be carried silently in cells for years, being passed on during ordinary mitosis Activation of HIV life cycle destroys THelper cells Weakened immune response to all foreign invaders, benign or aggressive

Pathologies: Autoimmune Diseases

Multiple Sclerosis (MS) myelin sheath (white matter) attacked and destroyed Myasthenia Gravis ACh receptors at neuro-muscular junction of skeletal muscle attacked and destroyed Graves Disease thyroid cells TSH receptor attacked and stimulated causing excess thyroid hormone (T3 & T4) production Type I Diabetes - destruction of pancreatic islet cells eliminates insulin secrection

Pathologies: Autoimmune Diseases

Systemic Lupus Erythematosus (SLE) generalized attack on connective tissues and nuclear antigens Glomerulonephritis - destruction of the glomerular capillaries causes impaired renal function Rheumatoid Arthritis - destruction of the synovial membranes in joints

Pathologies: Cancer
The immune system probably evolved first to respond to cancer cells
when a new cancer cell develops, new surface marker proteins (tumor antigens) often appear if the immune system recognizes these new surface markers as non-self, it will destroy the cell expressing them this immune surveillance is most effective in eliminating virus-induced tumor cells because they tend to express viral antigens which are not self

Leukemias and Lymphomas cancers of leukocytes

Pathologies: Hypersensitivities
Immediate hypersensitivities (allergies)
First exposure merely sensitizes one to an allergen (penicillin, venoms, dust, mold, pollen, etc.)
APCs digest and inappropriately present the allergen Subclones of B cells secreting IgE predominate in response Anti-allergen IgE attaches to mast cells and basophils

Later exposures produce dramatic responses

Antigen binds to IgE on mast cells and basophils Ag-IgE binding triggers these cells to release much histamine and other inflammatory molecules Local reactions swelling, rashes, erythema, itching Systemic reactions asthma, anaphylactic shock, death

Pathologies: Subacute Hypersensitivities

Caused by IgG and IgM Occurs 1-3 hr after exposure and lasts 10-15 hr Cytotoxic reactions
Ab bind to Ag on specific cells causing phagocytosis and complement-activated lysis May occur after transfusion of mismatched blood

Immune-complex hypersensitivities
Ags are widely distributed or insoluble Ag-Ab complexes cant be removed Intense inflammation Severe damage to local tissue Also involved in autoimmune diseases

Pathologies: Delayed Hypersensitivities

Occurs 1-3 days after exposure Cell-mediated immune response Causes mild swelling to serious cytotoxic tissue damage (contact dermatitis, e.g., TB skin test, poison ivy, latex gloves, etc.) [Note: Sometimes allergies may be temporarily transferred by blood or plasma transfusions.]

End Chapter 21