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C5

C4b 2a C3b

C5b

Microbe

Microbe

Fig. 8.11 The functions of complement. A, C3b opsonizes microbes and is recognized by the type 1

complement receptor (CR1) of phagocytes, resulting in ingestion and intracellular killing of the opsonized

microbes. Thus C3b is an opsonin. CR1 also recognizes C4b, which may serve the same function. Other complement products, such as the inactivated form of C3b (iC3b), also bind to microbes and are recognized by

other receptors on phagocytes (e.g., type 3 complement receptor, a member of integrin family of proteins). B,

Membrane attack complex creates pores in cell membranes and induces osmotic lysis of the cells. C, Small

peptides released during complement activation bind to receptors on neutrophils and other leukocytes and

stimulate inflammatory reactions. The peptides that serve this function are mainly C5a and C3a, released by

proteolysis of C5 and C3, respectively.

170 CHAPTER 8 Effector Mechanisms of Humoral Immunity

and antibody production. When C3 is activated by a

microbe by the alternative pathway, one of its breakdown products, C3d, is recognized by complement

receptor type 2 (CR2) on B lymphocytes. Signals delivered by this receptor enhance B cell responses against

the microbe. This process is described in Chapter 7

(see Fig. 7.5A) and is an example of an innate immune

response to a microbe (complement activation) enhancing an adaptive immune response to the same microbe

(B cell activation and antibody production). Complement proteins bound to antigen-antibody complexes are

recognized by follicular dendritic cells in germinal centers, allowing the antigens to be displayed for further B

cell activation and selection of high-affinity B cells. This

complement-dependent antigen display is another way

in which the complement system promotes antibody

production.

Inherited deficiencies of complement proteins result

in immune deficiencies and, in some cases, increased

incidence of autoimmune disease. Deficiency of C3

results in increased susceptibility to bacterial infections

that may be fatal early in life. Deficiencies of the early

proteins of the classical pathway, C2 and C4, may have

no clinical consequence, may result in increased susceptibility to infections, or are associated with an increased

incidence of systemic lupus erythematosus, an immune

complex-mediated autoimmune disease. The increased

incidence of lupus may be because the classical pathway

functions to eliminate immune complexes from the circulation, and these complexes accumulate in individuals

lacking C2 and C4. In addition, complement deficiencies may lead to defective signaling in B cells and a

failure of B cell tolerance (see Chapter 9). Deficiencies

of C9 and MAC formation result in increased susceptibility to Neisseria infections. Some individuals inherit

polymorphisms in the gene encoding MBL, leading to

production of a protein that is functionally defective;

such defects are associated with increased susceptibility to infections. Inherited deficiency of the alternative

pathway protein properdin also causes increased susceptibility to bacterial infection.

Regulation of Complement Activation

Mammalian cells express regulatory proteins that

inhibit complement activation, thus preventing complement-mediated damage to host cells (Fig. 8.12).

Many such regulatory proteins have been described,

and defects in these proteins are associated with

clinical syndromes caused by uncontrolled complement

activation.

• A regulatory protein called C1 inhibitor (C1 INH)

stops complement activation early, at the stage of

C1 activation. Deficiency of C1 INH is the cause of

a disease called hereditary angioedema. C1 INH is

a serine protease inhibitor that functions as a major

physiologic inhibitor of the cleavage of kallikrein,

the precursor of the vasoactive molecule bradykinin. Therefore, C1 INH deficiency results not only in

increased complement activation but also increased

proteolytic activation of bradykinin, and this is the

main reason for the vascular changes that lead to

leakage of fluid (edema) in many tissues.

• Decay-accelerating factor (DAF) is a glycolipid-linked cell surface protein that disrupts the binding of Bb to C3b and the binding of C4b to C2a, thus

blocking C3 convertase formation and terminating

complement activation by both the alternative and

the classical pathways. A disease called paroxysmal nocturnal hemoglobinuria results from the

acquired deficiency in hematopoietic stem cells of

an enzyme that synthesizes the glycolipid anchor for

several cell-surface proteins, including the complement regulatory proteins DAF and CD59. In these

patients, unregulated complement activation occurs

on erythrocytes, leading to their lysis.

• A plasma enzyme called Factor I cleaves C3b into

inactive fragments, with membrane cofactor protein

(MCP) and the plasma protein Factor H serving as

cofactors in this enzymatic process. Deficiency of

the regulatory proteins Factors H and I results in

increased complement activation and reduced levels

of C3 because of its consumption, causing increased

susceptibility to infection. Mutations in Factor H that

compromise its binding to cells are associated with a

rare genetic disease called atypical hemolytic uremic

syndrome, in which there are clotting, vascular, and

renal abnormalities. Certain genetic variants of Factor H are linked to an eye disease called age-related

macular degeneration.

These regulatory proteins are made by vertebrate host

cells but not by microbes. Because microbes lack these

regulatory proteins, the complement system can be activated on microbial surfaces much more effectively than

on normal host cells. Even in vertebrate cells, the regulation can be overwhelmed by too much complement

activation. For instance, host cells can become targets

CHAPTER 8 Effector Mechanisms of Humoral Immunity 171

DAF

C1q binds to

antigen-complexed

antibodies, resulting

in activation

of C1r2s2

Antibody

C1q C1r2s2

C1 INH

C1 INH

prevents C1r2s2

from becoming

proteolytically active

Formation of

C3 convertases

Dissociation of C3

convertases by DAF

C1r2s2

A

B

C3bBb C3b

Bb

Bb

C4b 2a C4b

2a

2a

Fig. 8.12 Regulation of complement activation. A, C1 inhibitor (C1 INH) prevents the assembly of the

C1 complex, which consists of C1q, C1r, and C1s proteins, thereby blocking complement activation by the

classical pathway. B, The lipid-linked cell surface protein decay-accelerating factor (DAF) and the type 1 complement receptor (CR1) interfere with the formation of the C3 convertase by blocking the binding of Bb (in the

alternative pathway) or C2a (in the classical pathway). Membrane cofactor protein (or CD46) and CR1 serve

as cofactors for cleavage of C3b by a plasma enzyme called factor I, thus destroying any C3b that may be

formed (not shown).

of complement if they are coated with large amounts

of antibodies, as in some hypersensitivity diseases (see

Chapter 11).

FUNCTIONS OF ANTIBODIES AT SPECIAL

ANATOMIC SITES

The effector mechanisms of humoral immunity described

so far may be active at any site in the body to which antibodies gain access. As mentioned previously, antibodies are

produced in peripheral lymphoid organs and bone marrow

and readily enter the blood, from which they may go anywhere. Antibodies also serve vital protective functions at

two special anatomic sites: the mucosal organs and the fetus.

Mucosal Immunity

Immunoglobulin A (IgA) is produced in mucosal

lymphoid tissues, transported across epithelia, and

binds to and neutralizes microbes in the lumens of

the mucosal organs (Fig. 8.13). Microbes often are

inhaled or ingested, and antibodies that are secreted

into the lumens of the respiratory or gastrointestinal

tract bind to these microbes and prevent them from

colonizing the host. This type of immunity is called

mucosal immunity (or secretory immunity). The principal class of antibody produced in mucosal tissues

is IgA. In fact, IgA accounts for about two-thirds of

the approximately 3 g of antibody produced daily by

a healthy adult, reflecting the vast surface area of the

intestines. The propensity of B cells in mucosal epithelial

tissues to produce IgA is because the cytokines that

induce switching to this isotype, including transforming growth factor ß (TGF-ß), are produced at high levels in mucosa-associated lymphoid tissues. In addition,

IgA-producing B cells that are generated in regional

lymph nodes or spleen tend to home to mucosal tissues

172 CHAPTER 8 Effector Mechanisms of Humoral Immunity

Protein Plasma

concentration

Function

C1 inhibitor

(C1 INH)

Factor I

Factor H

C4 binding

protein (C4BP)

Plasma proteins

200 µg/ml

35 µg/ml

480 µg/ml

300 µg/ml

Inhibits C1r and C1s serine

protease activity

Proteolytically cleaves

C3b and C4b

Causes dissociation of

alternative pathway

C3 convertase subunits

Co-factor for

Factor I-mediated

cleavage of C3b

Causes dissociation of

classical pathway

C3 convertase subunits

Co-factor for

Factor I-mediated

cleavage of C4b

Protein Distribution Function

Membrane

co-factor protein

(MCP, CD46)

Decay

accelerating

factor (DAF)

CD59

Type 1

complement

receptor

(CR1, CD35)

Membrane proteins

Leukocytes,

epithelial cells,

endothelial cells

Blood cells,

endothelial cells,

epithelial cells

Blood cells,

endothelial cells,

epithelial cells

Mononuclear

phagocytes,

neutrophils,

B and T cells,

erythrocytes,

eosinophils,

FDCs

Co-factor for

Factor I-mediated

cleavage of C3b and C4b

Blocks formation of

C3 convertase

Blocks C9 binding and

prevents formation

of the MAC

Causes dissociation of

C3 convertase subunits

Co-factor for

Factor I-mediated

cleavage of C3b and C4b

C

Fig. 8.12, cont’d C, The major regulatory proteins of the complement system and their functions. FDCs,

Follicular dendritic cells; MAC, membrane attack complex.

CHAPTER 8 Effector Mechanisms of Humoral Immunity 173

in response to chemokines produced in these tissues.

Also, some of the IgA is produced by a subset of B cells,

called B-1 cells, best studied in rodents, which also

have a propensity to migrate to mucosal tissues; these

cells secrete IgA in response to nonprotein antigens,

without T cell help.

Intestinal mucosal B cells are located in the lamina

propria, beneath the epithelial barrier, and IgA is produced in this region. To bind and neutralize microbial

pathogens in the lumen before they invade, the IgA

must be transported across the epithelial barrier into

the lumen. Transport through the epithelium is carried

out by a special Fc receptor, the poly-Ig receptor, which

is expressed on the basal surface of the epithelial cells.

This receptor binds IgA, endocytoses it into vesicles,

and transports it to the luminal surface. Here the receptor is cleaved by a protease, and the IgA is released into

the lumen still carrying a portion of the bound poly-Ig

receptor (the secretory component). The attached secretory component protects the antibody from degradation

by proteases in the gut. The antibody can then recognize

microbes in the lumen and block their binding to and

entry through the epithelium. IgA-mediated mucosal

immunity is the mechanism of protection from poliovirus infection that is induced by oral immunization with

the attenuated virus.

The gut contains a large number of commensal

bacteria that are essential for basic functions such as

absorption of food and, therefore, have to be tolerated

by the immune system. IgA antibodies are produced

mainly against potentially harmful and proinflammatory bacteria, thus blocking their entry through the gut

epithelium. Harmless commensals are tolerated by the

immune system of the gut by mechanisms that are discussed in Chapter 9.

Neonatal Immunity

Maternal antibodies are transported across the placenta to the fetus and across the gut epithelium of

neonates, protecting the newborn from infections.

Newborn mammals have incompletely developed

immune systems and are unable to mount effective

immune responses against many microbes. During their

early life, they are protected from infections by antibodies acquired from their mothers. This is an example of

naturally occurring passive immunity. Neonates acquire

maternal antibodies by two routes. During pregnancy,

maternal IgG binds to the FcRn expressed in the placenta, and is transported into the fetal circulation.

After birth, infants ingest maternal IgA antibodies that

are secreted into their mothers’ colostrum and milk.

Ingested IgA antibodies provide mucosal immune protection to the neonate. Thus, neonates acquire the antibody profiles of their mothers and are protected from

infectious microbes to which the mothers were exposed

or vaccinated.

IgAproducing

plasma cell

J chain

Dimeric

IgA

Poly-Ig

receptor with

bound IgA Secreted

IgA

Proteolytic

cleavage

Endocytosed

complex of

IgA and polyIg receptor

Microbe

Lamina propria Mucosal epithelial cell Lumen

Fig. 8.13 Transport of immunoglobulin A (IgA) through epithelium. In the mucosa of the gastrointestinal

and respiratory tracts, IgA is produced by plasma cells in the lamina propria and is actively transported through

epithelial cells by an IgA-specific Fc receptor, called the poly-Ig receptor because it recognizes IgM as well.

On the luminal surface, the IgA with a portion of the bound receptor is released. Here the antibody recognizes

ingested or inhaled microbes and blocks their entry through the epithelium.

174 CHAPTER 8 Effector Mechanisms of Humoral Immunity

EVASION OF HUMORAL IMMUNITY BY

MICROBES

Microbes have evolved numerous mechanisms to evade

humoral immunity (Fig. 8.14). Many bacteria and viruses

mutate their antigenic surface molecules so that they can no

longer be recognized by antibodies produced in response to

the original microbe. Antigenic variation typically is seen in

viruses, such as influenza virus, human immunodeficiency

virus (HIV), and rhinovirus. HIV mutates its genome at

a high rate, and therefore different strains contain many

variant forms of the major antigenic surface glycoprotein of

HIV, called gp120. As a result, antibodies against exposed

determinants on gp120 in any one HIV subtype may not

protect against other virus subtypes that appear in infected

individuals. This is one reason why gp120 vaccines are not

effective in protecting people from HIV infection. Bacteria such as Escherichia coli vary the antigens contained in

their pili and thus evade antibody-mediated defense. The

trypanosome that causes sleeping sickness expresses new

surface glycoproteins whenever it encounters antibodies

against the original glycoprotein. As a result, infection with

this protozoan parasite is characterized by waves of parasitemia, each wave consisting of an antigenically new parasite that is not recognized by antibodies produced against

the parasites in the preceding wave. Other microbes inhibit

complement activation, or resist opsonization and phagocytosis by concealing surface antigens under a hyaluronic

acid capsule.

VACCINATION

Now that we have discussed the mechanisms of host

defense against microbes, including cell-mediated immunity in Chapter 6 and humoral immunity in this chapter,

it is important to consider how these adaptive immune

responses can be induced with prophylactic vaccines.

Vaccination is the process of stimulating protective

adaptive immune responses against microbes by exposure to nonpathogenic forms or components of the

microbes. The development of vaccines against infections

has been one of the great successes of immunology. The only

human disease to be intentionally eradicated from the earth

is smallpox, and this was achieved by a worldwide program

Mechanism of

immune evasion

Example(s)

Antigenic

variation

Inhibition of

complement

activation

Blocking by

hyaluronic acid

capsule

Many viruses (e.g.,

influenza, HIV)

Bacteria (e.g.,

Neisseria gonorrhoeae,

Escherichia coli)

Protozoa (e.g.,

Trypanosoma cruzi)

Many bacteria

Streptococcus

Fig. 8.14 Evasion of humoral immunity by microbes. This figure shows some of the mechanisms by

which microbes evade humoral immunity, with illustrative examples. HIV, Human immunodeficiency virus.

CHAPTER 8 Effector Mechanisms of Humoral Immunity 175

of vaccination. Polio is likely to be the second such disease,

and as mentioned in Chapter 1, many other diseases have

been largely controlled by vaccination (see Fig. 1.2).

Several types of vaccines are in use and being developed (Fig. 8.15).

• Some of the most effective vaccines are composed of

attenuated microbes, which are treated to abolish pathogenicity while retaining their infectivity and antigenicity.

Immunization with these attenuated microbes stimulates the production of neutralizing antibodies against

microbial antigens that protect vaccinated individuals

from subsequent infections. For some infections, such

as polio, the vaccines are given orally to stimulate mucosal IgA responses that protect individuals from natural

infection, which occurs by the oral route.

• Vaccines composed of microbial proteins and polysaccharides, called subunit vaccines, work in the same way.

Some microbial polysaccharide antigens (which cannot

stimulate T cell help) are chemically coupled to proteins

so that helper T cells are activated and high-affinity

antibodies are produced against the polysaccharides.

These are called conjugate vaccines, and they are excellent examples of the practical application of our knowledge of helper T cell–B cell interactions (see Chapter 7).

Immunization with inactivated microbial toxins and

with microbial proteins synthesized in the laboratory

stimulates antibodies that bind to and neutralize the

native toxins and the microbes, respectively.

One of the continuing challenges in vaccination is to

develop vaccines that stimulate cell-mediated immunity

against intracellular microbes. Injected or orally administered antigens are extracellular antigens, and they induce

mainly antibody responses. Many newer approaches are

being tried to stimulate cell-mediated immunity by vaccination. One of these approaches is to incorporate microbial

antigens into viral vectors, which will infect host cells and

produce the antigens inside the cells. Another technique is

to immunize individuals with DNA encoding a microbial

antigen in a bacterial plasmid. The plasmid is ingested by

host APCs, and the antigen is produced inside the cells.

Many of these strategies have been successfully tested in

animal models, but few have shown clinical efficacy to date.

Type of vaccine Examples Form of protection

Live attenuated,

or killed, bacteria

Live attenuated

viruses

Subunit (antigen)

vaccines

Conjugate

vaccines

Haemophilus

influenzae,

Streptococcus

pneumoniae

(pneumococcus)

Hepatitis virus

(recombinant proteins)

Clinical trials have

been done

Clinical trials ongoing

for several infections

Antibody response

Antibody response;

cell-mediated immune

response

Antibody response

Helper T cell–

dependent antibody

response to

polysaccharide

antigens

Synthetic

vaccines

Viral vectors

DNA vaccines

BCG, cholera

Polio, rabies

Tetanus toxoid,

diphtheria toxoid

Antibody response

Cell-mediated and

humoral immune

responses

Cell-mediated and

humoral immune

responses

Fig. 8.15 Vaccination strategies. A summary of different types of vaccines in use or tried, as well as the

nature of the protective immune responses induced by these vaccines. BCG, Bacille Calmette-Guérin; HIV,

human immunodeficiency virus.

176 CHAPTER 8 Effector Mechanisms of Humoral Immunity

SUMMARY

• Humoral immunity is the type of adaptive immunity that is mediated by antibodies. Antibodies prevent infections by blocking the ability of microbes

to invade host cells, and they eliminate microbes by

activating several effector mechanisms.

• In antibody molecules, the antigen-binding (Fab)

regions are spatially separate from the effector (Fc)

regions. The ability of antibodies to neutralize microbes

and toxins is entirely a function of the antigenbinding regions. Even Fc-dependent effector functions

are activated only after antibodies bind antigens.

• Antibodies are produced in lymphoid tissues and

bone marrow, but they enter the circulation and are

able to reach any site of infection. Heavy-chain isotype switching and affinity maturation enhance the

protective functions of antibodies.

• Antibodies neutralize the infectivity of microbes and

the pathogenicity of microbial toxins by binding to

and interfering with the ability of these microbes and

toxins to attach to host cells.

• Antibodies coat (opsonize) microbes and promote their

phagocytosis by binding to Fc receptors on phagocytes.

The binding of antibody Fc regions to Fc receptors also

stimulates the microbicidal activities of phagocytes.

• The complement system is a collection of circulating and cell surface proteins that play important

roles in host defense. The complement system may

be activated on microbial surfaces without antibodies (alternative and lectin pathways, mechanisms of

innate immunity) and after the binding of antibodies

to antigens (classical pathway, a mechanism of adaptive humoral immunity).

• Complement proteins are sequentially cleaved, and

active components, in particular C4b and C3b,

become covalently attached to the surfaces on which

complement is activated. The late steps of complement

activation lead to the formation of the cytolytic MAC.

• Different products of complement activation promote phagocytosis of microbes, induce cell lysis, and

stimulate inflammation. Mammals express cell surface and circulating regulatory proteins that prevent

inappropriate complement activation on host cells.

• IgA antibody is produced in the lamina propria of

mucosal organs and is actively transported by a special Fc receptor across the epithelium into the lumen,

where it blocks the ability of microbes to invade the

epithelium.

• Neonates acquire IgG antibodies from their mothers through the placenta, using the FcRn to capture

and transport the maternal antibodies. Infants also

acquire IgA antibodies from the mother’s colostrum

and milk by ingestion.

• Microbes have developed strategies to resist or evade

humoral immunity, such as varying their antigens and

becoming resistant to complement and phagocytosis.

• Most vaccines in current use work by stimulating the

production of neutralizing antibodies.

REVIEW QUESTIONS

1. What regions of antibody molecules are involved in

the functions of antibodies?

2. How do heavy-chain isotype (class) switching and

affinity maturation improve the ability of antibodies

to combat infectious pathogens?

3. In what situations does the ability of antibodies to

neutralize microbes protect the host from infections?

4. How do antibodies assist in the elimination of

microbes by phagocytes?

5. How is the complement system activated?

6. Why is the complement system effective against

microbes but does not react against host cells and

tissues?

7. What are the functions of the complement system,

and what components of complement mediate these

functions?

8. How do antibodies prevent infections by ingested

and inhaled microbes?

9. How are neonates protected from infection before

their immune system has reached maturity?

Answers to and discussion of the Review Questions are

available at Student Consult.

177

Self–Nonself Discrimination in the

Immune System and Its Failure

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