Chapter 21

Nonspecific Body Defenses and Immunity

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

Defense Systems
1. Innate (nonspecific) defense
External body membranes Inflammation
Antimicrobial proteins, phagocytes and other cells

2. Adaptive (specific) defense

T cells and B cells

Innate Defense System

Surface Barriers
First line of defense Skin and mucosal membranes Mechanical and chemical protection
Second line of defense Phagocytes Natural Killer cells (NK lymphocytes) Inflammation Antimicrobial proteins Fever

1. 2. 3. 4. 5.

Internal Nonspecific Defenses

Innate Defense: Surface Barriers

Skin and mucosal membranes
Mechanical protection
Intact epidermis Mucous membranes
line body cavities, mucus prevents drying, traps foreign things nose hairs, respiratory tract cilia

Lacrimal apparatus -- tear glands and ducts

continually wash the eye to dilute microbial growth

Saliva - dilute microbes on the teeth, tongue, and gums Urine - flow dilutes, and acid pH helps kill, microorganisms Defecation and vomiting - expel toxic microbes

Innate Defense: Surface Barriers

Chemical protection: helps prevents 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

Macrophage are the chief phagocytic cell Derived from monocytes

Free macrophages wander throughout a region in search of cellular debris

Kupffer cells (liver) and microglia (brain) are fixed macrophages Neutrophils become phagocytic when encountering infectious material Eosinophils are weakly phagocytic, deploy destructive granules against parasitic worms

Mast cells bind and ingest a wide range of bacteria

Mechanism of Phagocytosis
Chemotaxis Adherence recognition of carbohydrate signature
Aided by opsonization

Ingestion Digestion
Respiratory burst

Innate Defense: Natural Killer Cells

Natural Killer cells (NK lymphocytes)
Small, distinct group of large granular lymphocytes Nonspecific killers which respond to the lack of selfantigens and to the presence of specific sugars before the antigen-specific immune system is activated Able to kill virus-infected body cells and some tumor cells by releasing various defensive molecules (perforin and granzymes) not by phagocytosis Secrete potent chemicals 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

Inflammation
Edema - A surge of protein-rich fluid into tissue spaces
Helps to dilute harmful substances Brings in large quantities of oxygen and nutrients needed for repair Allows entry of clotting proteins, which prevents the spread of bacteria

Inflammation
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
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

Infectious granuloma
Macrophage resistant bacteria in macrophage are surrounded by healthy macrophage surrounded by fibrous capsule

http://medical-dictionary.thefreedictionary.com/abscessing

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

Innate Defense: Antimicrobial Proteins

Enhance the innate defenses by:
1. Attacking microorganisms directly 2. Hindering microorganisms ability to reproduce

The most important antimicrobial proteins are:

1. Interferon 2. Complement proteins 3. Transferrins bind Fe2+ in plasma, inhibiting bacterial growth

Interferon (IFN)
Produced by cells in response to viral infections
Diffuses to uninfected cells and binds to surface receptors
stimulates production of antiviral proteins like PKR.
PKR blocks viral replication

enhances macrophages and natural killer lymphocytes inhibits growth of virally infected cells suppresses growth of tumor cells

FDA-approved alpha IFN is used:

As an antiviral drug against hepatitis C virus To treat genital warts caused by the herpes virus

Innate Defense: Antimicrobial Proteins

Complement
20 plasma and cell membrane proteins that exist in inactive forms When activated, the complement system functions to complement or enhance certain immune, allergic, and inflammatory responses Amplifies all aspects of the inflammatory response Kills bacteria and certain other cell types (our cells are immune to complement) Enhances the effectiveness of both nonspecific and specific defenses

Complement Pathways
Classical pathway is linked to the immune system
Requires binding of antibodies to antigens of invading organisms Subsequently, C1 binds to the antigen-antibody complexes (complement fixation)

Alternative pathway is triggered by interaction of factors B, D, and P, with polysaccharide molecules present on microorganisms (direct

Complement Pathways
Each pathway involves a cascade Both pathways converge on C3, which cleaves into C3a and C3b C3a enhances inflammation C3b initiates formation of a membrane attack complex (MAC)
MAC causes cell lysis by interfering with a cells ability to control intracellular [Ca2+]

C3b also causes opsonization

Innate Defense: Fever

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

Innate Defense System: Review

Surface Barriers
First line of defense Skin and mucosal membranes Mechanical and chemical protection

Internal Nonspecific Defenses

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

Summary of the Primary Immune Response

 The adaptive immune system is a functional system that:

Acts to immobilize, neutralize, or destroy foreign substances Amplifies the inflammatory response and activates complement Is antigen-specific, systemic, and has memory
Recognizes specific foreign substances

Adaptive Defense

Has two separate but overlapping arms

Humoral, or antibody-mediated immunity Cellular, or cell-mediated immunity

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 antibody production
Reactivity: the ability to react with the products of the activated lymphocytes and the antibodies released in response to them

Adaptive Defense
Specificity and memory differentiate the adaptive system from the non-specific system
Specificity: the antigen triggers immune defenses (from lymphocytes) 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

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.

Adaptive Defense
Chemical nature of antigens
Complete: large, complex molecules usually proteins (nucleo-, lipo-, glyco-) -sometimes carbohydrates or lipids
Are immunogenic & reactive

Incomplete: smaller molecules (haptens)

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

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
1. Class I MHC found on all body cells except RBC's 2. Class II MHC - only on antigen presenting cells (APCs), thymus cells, and activated T cells

Cells of the Adaptive Immune System: Lymphocytes

Immature lymphocytes released from bone marrow are essentially identical Whether a lymphocyte matures into a B cell or a T cell depends on where it becomes able to recognize a specific antigen (immunocompetent)
B cells mature in the bone marrow & oversee humoral immunity T cells mature in the thymus & oversee cell-mediated immunity

Immunocompetent B or T cells
Display a unique type of receptor for a specific antigen
Occurs before cells encounter antigens they may later attack It is genes, not antigens, that determine which foreign substances our immune system will recognize and resist

Naive cells are exported to secondary lymphoid tissue where cells may encounter antigens
Mature into fully functional antigen-activated cells upon binding with their recognized antigen

T Cells
T cells mature in the thymus under positive and negative selection pressures
Positive selection thymic cortex
Selects T cells with a weak response to selfantigens (MHC molecules) These cells become both immunocompetent and self-tolerant Non-selected cells die via apoptosis
Apoptosis

Survive

Negative selection inner portion of thymic cortex

Eliminates T cells that strongly react with self-antigens (other than the MHC)

Apoptosis

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 modify the self-reactive antigen (receptor editing)

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

Cells of the Adaptive Immune System: APCs

Antigen-presenting cells (APCs):
Engulf, process, and present parts of antigens Do not respond to specific antigens Play essential auxiliary roles in immunity Dendritic (Langerhans) cells: connective tissue and epidermis Macrophage B lymphocytes

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

Humoral Immune Response

Antigen challenge first encounter between an antigen and a naive lymphocyte
Usually in the spleen or lymph node, but can occur in other lymphoid organs

If the lymphocyte is a B cell, a humoral immune response is provoked

1. Binding of the antigen to the receptor activates the lymphocyte 2. Clonal selection occurs

Clonal Selection
Activated B cell grows & mitotically divides, forming clones bearing the same antigenspecific receptors

Fate of the Clones

Most clone cells become plasma cells that secrete specific antibodies

Other clones become memory cells that can respond to subsequent exposures of the same antigen

Immunological Memory
Primary immune response cellular differentiation and proliferation, which occurs on the first exposure to a specific antigen
Lag period: 3 to 6 days after antigen challenge Peak levels of plasma antibody are achieved in 10 days Antibody levels then decline

Immunological Memory
Secondary immune response re-exposure to the same antigen
Sensitized memory cells respond within hours Antibody levels peak in 2 to 3 days at much higher levels than in the primary response Antibodies bind with greater affinity, and their levels in the blood can remain high for weeks to

Primary and Secondary Humoral Responses

Soluble antibodies are the simplest ammunition of the immune response and interact in extracellular environments

Types of Humoral Immunity

Active immunity: B cells encounter antigens and produce antibodies against them
Naturally acquired response to a bacterial or viral infection Artificially acquired response to a vaccine of dead or attenuated (weakened) pathogens

Passive immunity: B cells are not challenged by antigens

Immunological memory does not occur Protection ends when antigens naturally degrade in the body Naturally acquired from the mother to her fetus via the

Antibodies
Also called immunoglobulins
Constitute the gamma globulin portion of blood proteins

Are soluble proteins secreted by activated B cells and plasma cells in response to an antigen
Are capable of binding specifically with that antigen

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

Antibody Structure
Antibodies responding to different antigens have different V regions but the C region is the same for all antibodies in a given class

C regions form the stem of the Y-shaped antibody and:

Serve common functions in all antibodies Dictate the cells and chemicals that the antibody can bind to Determine the class of the antibody Determine how the antibody class will function in

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 response; crosses the placenta and confers passive immunity IgA: dimer that helps prevent attachment of pathogens to epithelial cell surfaces
IgE: monomer that binds to mast cells and basophils, causing histamine release when activated

Antibody Targets
Antibodies themselves do not destroy antigen; they inactivate and tag it for destruction All antibodies form an antigen-antibody (immune) complex

Defensive mechanisms used by antibodies are neutralization, agglutination, precipitation, 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

2. Agglutination: Antibodies bind the same determinant on more than one antigen
Makes antigen-antibody complexes that are cross-linked into large lattices Cell-bound antigens are cross-linked, causing clumping (agglutination)

3. Precipitation soluble molecules are cross-linked

Antibody Mechanisms of Action

4. Complement fixation is the main mechanism used against cellular antigens
Antibodies bound to cells change shape and expose complement binding sites This triggers complement fixation and cell lysis Complement activation:
Enhances the inflammatory response Uses a positive feedback cycle to promote phagocytosis

Enlists more and more defensive elements

Mechanisms of Antibody Action

Figure 21.13

Cell-Mediated Immunity
Since antibodies are useless against intracellular antigens, cell-mediated immunity is needed Two major populations of T cells mediate cellular immunity
CD4 cells (T4 cells) are primarily helper T cells (TH) CD8 cells (T8 cells) are cytotoxic T cells (TC) that destroy cells harboring foreign antigens

Other types of T cells are:

Suppressor T cells (TS) Memory T cells

Importance of Cell-Mediated Immunity

T cells recognize and respond only to processed fragments of antigen displayed on the surface of body cells T cells are best suited for cell-to-cell interactions, and target:
Cells infected with viruses, bacteria, or intracellular parasites Abnormal or cancerous cells Cells of infused or transplanted foreign tissue

Cell -Mediated Immunity

Basic steps
1. Recognition of antigen presented by an antigenpresenting cell by T cell receptors (TCRs)

2. Co-stimulation of the T cell

3. Production of a clone of identical effector T cells capable of recognizing initial activator (antigen) 4. Elimination of the foreign intruder

T Cell Activation- Step 1: Antigen Binding

Mobile APCs (Langerhans/dendritic cells) quickly alert the body to the presence of antigen by migrating to the lymph nodes and presenting antigen
May display exogenous antigens from dying virusinfected cells or tumor cells on class I and II MHC proteins

T cell antigen receptors (TCRs):

Activated by binding to an antigen-MHC protein complex Have variable and constant regions consisting of two chains (alpha and beta) Linked to multiple intracellular signaling pathways

T Cell Activation- Step 2: Co-stimulation

T cells must bind to other surface receptors on an APC
Macrophage & dendritic cells produce surface B7 proteins when nonspecific defenses are mobilized B7 binding with the CD28 receptor on the surface of T cells is a crucial co-stimulatory signal

After co-stimulation, cytokines such as interleukin 1 and 2 stimulate proliferation and differentiation of T cells

Cell-Mediated Immunity
Primary T cell response peaks within a week after signal exposure T cells then undergo apoptosis between days 7 and 30 Effector activity wanes as the amount of antigen declines

The disposal of activated effector cells is a protective mechanism for the body
Memory T cells remain and mediate

Helper T Cells (TH)

Regulatory cells that play a central role in the immune response Once primed by APC presentation of antigen, they:
Chemically or directly stimulate proliferation of other T cells Stimulate B cells that have already become bound to antigen

There is NO immune response without TH

Helper T Cell
TH cells interact directly with B cells that have antigen fragments on their surfaces bound to MHC II receptors TH cells stimulate B cells to divide more rapidly and begin antibody formation

B cells may be activated without TH cells by binding to T cellindependent antigens

Most antigens, however, require TH co-stimulation to activate B cells Cytokines released by TH amplify nonspecific defenses

Cytotoxic T Cell (Tc)

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 or transplants

Cytotoxic T Cells
Bind to self-antiself complexes on all body cells Infected or abnormal cells can be destroyed as long as appropriate antigen and costimulatory stimuli In contrast, natural killer cells activate their killing machinery when they bind to MICA receptor
MICA receptor MHC-related cell surface protein

Summary of the Primary Immune Response

Adaptive Immunity: Summary

Two-fisted defensive system that uses lymphocytes, APCs, and specific molecules to identify and destroy nonself particles Its response depends upon the ability of its cells to:
Recognize foreign substances (antigens) by binding to them Communicate with one another so that the whole system mounts a response specific to those

Adaptive Immunity: Summary

To start an immune response, B and T cells must recognize foreign antigen B cells can recognize and bind to certain antigens in the blood or the extracellular fluid (ECF) More often, B and T cells only recognize antigen (protein fragments) when Ag is presented by the phagocytes in combination with MHC Class II surface markers