The Complement System

Objective:
• Definition of Complement
The Complement System
• Biologic Effects of Complement
• Complement Pathways
• Regulation of the Complement System
• Complement Deficiencies and Pathogen Evasion

Dr. Suzan Yousif

COMPLEMENT
- The complement system includes serum and membrane bound proteins that function in both innate and
adaptive host defense systems.

- These proteins are highly regulated and interact via a series of proteolytic cascades.
- Several complement components are proenzymes, which must be cleaved to form active enzymes.
- The components of the classical pathway are numbered from C1 to C9, and the reaction sequence is C1-C4-
C2-C3-C5-C6-C7-C8-C9.

Biologic Effects of Complement

Activation of complement results in four major outcomes:
(1) cytolysis 5
phigrise is
->
(2) chemotaxis
(3) opsonization -> Asosx,
(4) anaphylatoxins. -> allergy -> Ad
1. Cytolysis is the lysis of cells, such as bacteria, virus infected cells, and tumor cells. This process
Se
occurs through the development of the membrane attack complex (MAC) (C5b, 6, 7, 8, 9), which
inserts into the membrane of an organism or cell. The MAC leads to loss of osmotic integrity and cell
lysis.
2. Chemotaxis is a process in which an immune cell, usually a phagocyte, is attracted to and moves toward a
soluble factor. For example, C5a is a potent chemotactic agent that stimulates movement of neutrophils and
monocytes toward sites of antigen deposition.

3. Opsonization is a process in which microorganisms and antigen–antibody complexes become coated with
molecules that bind to receptors on phagocytes. Phagocytosis is more efficient in the presence of C3b
because of the presence of C3b receptors on phagocytes.

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4. Anaphylatoxins promote vasodilation and increased vascular permeability. Both C3a and C5a are potent

6 Is
promoters of vasodilation and vascular permeability. These two complement components also stimulate
mast cells and basophils to release histamine. This function of complement results in an increased blood
>
->
flow to the site of infection, allowing more complement, antibodies, and immune cells to enter the site of
infection.

peniclin; 9
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~
~

7
• Complement Pathways
• There are three major pathways that activate complement, the classical pathway,
alternative pathway, and mannan-binding lectin (MBL) pathway.

• Each of these pathways results in the formation of the Membrane Attack Complex
-
(MAC). I
. W -

- -

• All three pathways lead to the release of C5 convertase, which breaks down C5 into -
-

C5a and C5b.

• As mentioned previously, C5a is an anaphylatoxin as well as a chemotactic factor.

• C5b binds to C6 and C7 to form a complex that inserts into the membrane bilayer.

• C8 then binds to the C5b–C6–C7 complex followed by polymerization of up to 16 C9

molecules to produce the MAC.

• The MAC now generates a channel or pore in the membrane and causes cytolysis by
allowing free passage of water across the cell membrane.
• The Classical Pathway
• C1, which binds to the Fc region of an immunoglobulin, is composed of three proteins, C1q, C1r, and C1s. C1q is an
I

aggregate of polypeptides that bind to the Fc portion of IgG and IgM.

-
- -
-

• The antibody–antigen immune complex bound to C1 activates C1s, which cleaves C4 and C2 to form C4b2b. =>
-
-

• The latter is an active C3 convertase, which cleaves C3 molecules into two fragments, C3a and C3b. C3a is an
anaphylatoxin.

• C3b forms a complex with C4b2b, producing a new enzyme, C5 convertase, which cleaves C5 to form C5a and C5b.

• This leads to MAC and then to cell lysis.

• Only IgM and IgG activate or fix complement via the classical pathway. Furthermore, only IgG subclass 1, 2, and 3 fix

*
complement; IgG4 does not.
-

• An example of the classical complement pathway in action can be observed in Herpes simplex virus (HSV) infections. HSV
replication within cells is accompanied by the insertion of virus proteins on the cell surface. A specific antiHSV antibody
can bind to the surface of the infected cell by its Fab site. The Fc portion of the antigen–antibody complex is now exposed
and ready for C1 to attach. The classical pathway is now activated and the infected cell is destroyed by MAC
• The Alternative Pathway
• Infectious agents can activate the complement system
by triggering the cellular production of factors B, D, and
properdin. jid
4. B
• These factors cleave C3 and generate C3 convertase.
*
• C3 convertase (C3bBb) that was generated during the
alternative pathway produces more C3b.

• The additional C3b binds to the C3 convertase to form

C3bBbC3b. A

• This enzyme is the alternative pathway C5 convertase

that generates C5b, leading to production of the MAC
described earlier. I
• Mannan-Binding Lectin Pathway
• In recent years, the concept of an additional pathway of complement
activation has emerged, and a third pathway has been identified as the MBL
pathway. Its main constituent is a plasma protein MBL. I -

• MBL binds to sugar residues such as mannose found in microbial surface

2 X:G-bacteria

polysaccharides such as LPS. The MBL complex, when bound to microbial

surfaces, can activate C4 and C2.

• The remaining steps in the pathway are the same as the classic pathway of
complement activation.
 Regulation of the Complement System
• Several serum proteins regulate the complement system at different stages:

s
(1) C1 inhibitor binds to and inactivates the serine protease activity of C1r and C1s;
I -

(2) factor I cleaves C3b and C4b, thereby reducing the amount of C5 convertase available;
-
-

-
-

(3) factor H enhances the effect of factor I on C3b

-

(4) factor P (properdin) protects C3b and stabilizes the C3 convertase of the alternative pathway.

❖Regulation is also provided by proteins that have the ability to accelerate the decay of the
complement proteins, such as decay-accelerating factor, a membrane-bound protein found on
most blood cell surfaces that can act to accelerate dissociation of the C3 convertases of all three
pathways
• Complement Deficiencies and Pathogen Evasion
• Many genetic deficiencies of complement proteins have been described, and these generally lead to
enhanced susceptibility to infectious disease (eg,
• C2 deficiency frequently leads to serious pyogenic bacterial infections.
• Deficiency in components of the MAC greatly enhances susceptibility to neisserial infections.
• Deficiencies in components of the alternative pathway are also known (eg, properdin deficiency is
associated with greater susceptibility to meningococcal disease). -
• There are also deficiencies in complement regulating proteins. For example, lack of the C1 inhibitor protein
leads to hereditary angioedema.
• The complement system is an important host protective system. Some pathogens have evolved a
mechanism to evade it. For example, some microbes have developed surfaces that interfere with
opsonization by C3b or interfere with insertion of the MAC.
• Complement activation can also be inhibited by the presence of microbial generated proteins, such as,
protein A, and protein C, that bind IgG Fc. Finally, microbes can generate enzymes that degrade
complement components.
• Organisms that possess these inhibitory properties are usually more pathogenic.
 The Major Histocompatibility Complex

Objective:
• Definition of MHC
• Types of MHC
• Antigen Processing and Presentation

Dr. Suzan Yousif

• The major histocompatibility complex (MHC) was first detected as the genetic locus encoding the
-

glycoprotein molecules (transplantation antigens) responsible for the rapid rejection of tissue grafts
transplanted between genetically nonidentical individuals.

• It is now known that MHC

-
molecules bind peptide antigens and present them to T cells. Thus, these
transplantation antigens are responsible for antigen recognition by the T-cell receptor.

• In this respect, the T-cell receptor is different from antibody. Antibody molecules interact with antigen
directly, whereas the T-cell receptor only recognizes peptide antigens presented by MHC molecules on the
APC.

• The T-cell receptor is specific for antigen, but the antigen must be presented on a self-MHC molecule. ·
• The T-cell receptor is also specific for the MHC molecule. If the antigen is presented by another allelic ->
-

form of the MHC molecule in vitro (normally in an experimental situation), there is no recognition by the
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T-cell receptor. This phenomenon is known as MHC restriction.

• In humans, the MHC is a cluster of extensively studied genes located on chromosome 6. Among the many
important genes in the human MHC, also known as HLA, are those that encode the class I, class II, and class III
---

MHC proteins.

-
--- - - - -

---
...
-

Tymar nicrosisFactor
• MHC class I proteins are encoded by the HLA-A, -B, and -C genes.

• These proteins are made up of two chains:

(1) a transmembrane glycoprotein of MW 45,000, noncovalently

-
associated with
- --

~
(2) a non–MHC-encoded polypeptide of MW 12,000 that is known -

as  2-microglobulin.
RBC s
-
• Class I molecules are to be found on virtually all nucleated cells in
-

the body. Key exceptions are observed on cells in the retina and -

excep
->

brain. -
-
-

• The class I MHC locus also includes genes encoding proteins

involved in antigen processing (eg, transporters associated with
antigen processing [TAPs]).
• Class II proteins are encoded by the HLA-D region.
• there are three main families: the DP-, DQ-, and DR-encoded
-
-
-

molecules.
• This locus retains control of immune responsiveness and
different allelic forms of these genes confer striking differences
in the ability to mount an immune response against a given
dis

4
-
antigen.
- ~

• The HLA-D locus-encoded proteins are made up of two

noncovalently associated transmembrane glycoproteins of
-
approximately MW 33,000 and MW 29,000.
29533
-
• Unlike class I proteins, they have a restricted tissue distribution
-

and are chiefly found on macrophages, dendritic cells, B cells,

and other APCs. However, their expression on other cells (eg, -

endothelial cells or epithelial cells) is induced by IFN-. * -

• The genes of the MHC exhibit a remarkable genetic variability. The MHC is polygenic in that there are several genes for
-

each class of molecule. The MHC is also polymorphic. Thus, a large number of alleles exist in the population for each of
the genes. Each individual inherits a restricted set of alleles from his or her parent. Sets of MHC genes tend to be
inherited as a block or haplotype. There are relatively infrequent cross-over events at this locus.
18
#)
• Major histocompatibility class proteins show a broad specificity for peptide antigens, and many different
peptides can be presented by any given MHC allele (one peptide is bound at a time). The  helices that
-

form the binding cleft are the site of the amino acid residues that are polymorphic in MHC proteins (ie,
-

those that vary among alleles). This means that different alleles can bind and present different peptide
antigens. For all these reasons, MHC polymorphism has a major effect on antigen recognition.

• Analysis of the function of T cells with respect to interaction with MHC molecules reveals that peptide
antigens associated with class I MHC molecules are recognized by CD8-positive cytotoxic T lymphocytes,
whereas MHC class II–associated peptide antigens are recognized by CD4- positive helper T cells.

class 6 - CD
8
MHC
CD Y
MAC cls 2 ->
Antigen Processing and Presentation
• Antigen processing and presentation are the means by which antigens become associated with self-MHC
molecules for presentation to T cells with appropriate receptors.

-
• Proteins from exogenous antigens, such as bacteria, are internalized via endocytic vesicles into APCs such
as the various types of dendritic cells and macrophages. Then, as illustrated in Figure, they are exposed to
cellular proteases in intracellular vesicles. Peptides, approximately 10–30 amino acid residues in length, are
generated in endosomal vesicles. The endosomal vesicles can then fuse with exocytic vesicles containing
class II MHC molecules.
• The class II MHC molecules are synthesized, as are other membrane glycoproteins, in the rough
endoplasmic reticulum and then they proceed out through the Golgi apparatus. A third polypeptide, the
invariant chain (Ii), protects the binding site of the class II  dimer until the lowered pH of the
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compartment created after fusion with an endosomal vesicle causes a dissociation of the Ii chain.
• The MHC classII–peptide antigen complex is then transported to the cell surface for display and recognition
by a T-cell receptor of a CD4 T cell. The CD4 T cells can now provide help to other-
T cells and B cells.
-
2x:XX
• Several viruses attempt to defeat the immune response by interfering with the antigen-processing
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pathways. For example, an HIV Tat protein is able to inhibit expression of class I MHC molecules. A
herpesvirus protein binds to the TAPs, preventing transport of viral peptides into the endoplasmic
reticulum, where class I molecules are being synthesized. A consequence of these inhibitory
mechanisms is that the infected cells are not recognized by cytotoxic lymphocytes.

• Some superantigens are able to bind to MHC molecules outside the peptide-binding cleft. One
consequence is that whereas an individual peptide complexed to an MHC molecule will normally
stimulate only a small percentage of the T cells in an individual, superantigens cause up to 10% of T cells
to be nonspecifically activated. Examples of superantigens include certain bacterial toxins, including the
-

staphylococcal enterotoxins, toxic shock syndrome toxin, and group A streptococcal pyrogenic exotoxin
A. These antigens bind to the “outside” of the MHC protein and to the T-cell receptor. They are active at
-

very low concentrations (10–9 mol/L) and cause T cells expressing particular V
-
en
-
• sequences to be stimulated and to release large amounts of cytokines, including IL-1 and TNF. It is the
release of large amounts of cytokines (cytokine storm) from activated T lymphocytes that explain in part
the pathogenesis of diseases caused by organisms expressing superantigens.

• Understanding the details of antigen processing has helped to clarify our thinking about T-cell function.
An explanation for why T cells do not respond to carbohydrate antigens may be due to their inability to fit
well in the groove. Moreover, why T cells recognize only linear antigenic determinants may be explained
by the fact that T cells respond only to proteolytically processed antigen. Therefore, whether an antigen
is destined for class I or class II presentation depends on the intracellular compartments it traverses