Complement System

 Complement was discovered by Jules Bordet as a heat-labile component of normal plasma that
causes the opsonisation and killing of bacteria.
 Complement system is composed of more than 25 different proteins produced by hepatocytes,
macrophages and intestinal epithelial cells. Fibroblasts and intestinal epithelial cells make C1, while
the liver makes C3, C6, and C9.
 They are present in the circulation as inactive molecules/ zymogens (proenzymes).
 Complement proteins work in a cascade, where the binding of one protein promotes the binding of
the next protein in the cascade (activation of one protein enzymatically cleaves and activates the
next protein in the cascade).
 Complement components are numbered in the order in which they were discovered. During
activation, some complement components are split into two parts.
 The larger part of the molecule called "b" while the smaller fragment called "a" may diffuse away.
 In most cases it is the "b" fragment binds to the surface of the cell to be lysed (the fragments of C2
are an exception to this rule: C2a binds to the membrane while C2b is freed into serum or tissue
spaces).
 Inactivated fragments are indicated by a small "i". Enzymatically active forms are symbolized by a
bar over the letter or number.
 Activation of complement results in the production of several biologically active molecules, which
contribute to nonspecific immunity and inflammation.
 Complement is not antigen-specific and it is activated immediately in the presence of pathogen, so
it is considered part of innate immunity.
 Since antibody also activates some complement proteins, complement activation is also part of
humoral immunity.
Activation of the Complement System
 There are three ways to activate the complement system, involving different molecules initially but
converging to produce the same effector molecules.
 Each involves activation of enzymes that cleave their substrates to form a cascade, so that the
complement response is amplified.
1. The Classical Pathway
2. The Mannose-Binding Lectin Pathway
3. The Alternative Pathway
 All three pathways produce C3 convertase, an enzyme which triggers further effects downstream. The
effects of C3 convertase are discussed below.
Classical Pathway
This pathway involves complement components C1, C4 and C2. The pathway is triggered by
antibody-antigen complexes binding to C1, which itself has three subcomponents C1q, C1r and C1s.
The pathway forms a C3 convertase, C4b2a, which splits C3 into two fragments; the large fragment,
C3b, can covalently attach to the surface of microbial pathogens and opsonise them; the small
fragment, C3a, activates mast cells, causing the release of vasoactive mediators such as histamine.
Alternative Pathway
This pathway involves various factors, B, D, H & I, which interact with each other, and with C3b, to
form a C3 convertase, C3bBb, that can activate more C3, hence the pathway is sometimes called „the
amplification loop‟. Activation of the loop is promoted in the presence of bacterial and fungal cell
walls, but is inhibited by molecules on the surface of normal mammalian cells.
Mannose-binding Lectin Pathway
This pathway is activated by the binding of mannose-binding lectin (MBL) to mannose residues on
the pathogen surface. This in turn activates the MBL-associated serine proteases, MASP-1 and
MASP-2, which activate C4 and C2, to form the C3 convertase, C4b2a.
Lytic Pathway
This pathway is initiated by the splitting of C5, and attachment of C5b to a target. C6, C7, C8 and C9
unite with C5b, and this membrane-attack complex (MAC), when inserted into the outer membrane
of some bacteria, can contribute to their death by lysis. Red cells which have antibody bound to the
cell surface can also activate the classical and lytic pathways, and become susceptible to lysis.
 REGULATION OF COMPLEMENT ACTIVATION:
 C1 Inhibitor (also known as serpin) inhibits the production of C3b by combining with and
inactivating Cq1rs
complex. This prevents formation of the C3 convertase, C4b2b.
 Protein H inhibits the production of C3b by inhibiting the binding of Factor B to membrane-bound
C3b, thereby preventing cleavage of B to Bb and production of the C3 convertase, C3bBb. Factor I
inhibits the production of C3b by cleaving C3b into inactive forms.
 Regulatory proteins promote or inhibit complement activity and protect self cells from lysis. C5b67
can bind
indiscriminately to any cell membrane leading to their lysis. Such an indiscriminate damage to by-
standing cells is prevented by protein S (vitronectin) that binds to C5b67 complex and blocks its
indiscriminate binding to cells other than the primary target.
 Decay accelerating factor (DAF) accelerates breakdown of C3 convertase.
 Homologous restriction factor (HRF) prevents insertion of C8 and C9 into membranes.
Biological Effects Of Complement
A) Cytolysis [C5b6789] (Note: the bar identifies an activated complex)
Destruction of target cells by lysis of the cell membrane. This is termed cytotoxicity in the case of nucleated
cells, hemolysis for red blood cells, or bacteriolysis in the case of bacteria. (NOTE: Not all bacterial and
eukaryotic cells are susceptible to complement- dependent lysis).
B) Anaphylotoxin activity (= "vasoactive" or "phlogistic") [C3a, C5a]
Stimulation of mast cells to release histamine and other substances, resulting in increased capillary
permeability and local accumulation of fluid in the tissue.
C) Chemotaxis [C5a, C5b67]
Attraction of polymorphonuclear neutrophils (PMN's) to a local site of inflammation.
D) Opsonization (= "immune adherence") [C3b]
Facilitation of phagocytosis by macrophages or PMN's via cell-surface receptors specific for complement
components ("complement receptors", or "CRs")
 E) Tissue damage [C5b6789; PMN's]
Both the lytic complex and the inflammatory PMN's can cause considerable damage to normal tissues, for
instance in an Arthus Reaction or in Immune Complex Disease.
Biological Consequences of Complement Activation
i. Direct killing. The formation of functional MAC on cell surfaces causes cell lysis.
ii. Inflammation. Anaphylatoxins (C3a and C5a) increase vascular permeability, allowing influx of cells
into damaged tissue.
iii. Removal of necrotic and apoptotic cells. C3b deposits on dying cells, signaling rapid clearance by
phagocytic cells (neutrophils). Once a cell is dead, it can no longer be protected from opsonization.
iv. Clearance of immune complexes. C3b deposits on antibody-antigen complexes and causes their
dissociation and clearance.
v. Neutralization of viral infection. Complement on antibody-antigen complexes forms a thick coat, thus
blocking viral entry into cells.
vi. Enhancement of B-cells. During an infection, B-cells are activated by binding of C3b to complement
receptors.
Clearance of immune complexes, neutralization of viral infection, and enhancement of B-cells are all
consequences of the acquired system.
Deficiency Of Complement:
Deficiency defects are transmitted as phenotypically autosomal-recessive traits. Genetic deficiencies in the
alternative pathway are very rare. Deficiency of properdin is X-linked. Deficiency of the C1 inhibitor is
inherited as an autosomal dominant. All patients with complement deficiency are more or less unduly
susceptible to infection and to development of immune complex disease.

DEFICIENCY CLINICAL CORRELATION

Autoimmune collagen vascular disease, susceptibility to infectious
C3 (any pathway)
diseases
Classical pathway
Autoimmune collagen vascular disease, susceptibility to infectious
C1q, C1r, C1s, C4, C2
diseases
Alternate pathway
Factor B Meningococcemia
Factor D Recurrent pyogenic infections
Properdin Recurrent pyogenic infections, Meningococcemia
Lectin pathway Recurrent infections
Membrane attack complex
C5 Recurrent disseminated Neisseria infections, SLE
C6, C7, C8 Recurrent disseminated Neisseria infections
C9 Susceptibility to infectious diseases
C1 inhibitor Hereditary angionurotic oedema (Quincke's disease)
Factor I, Factor H Autoimmune collagen vascular disease, Recurrent pyogenic infections
CR1 SLE
DAF, HRF Paroxysmal nocturnal hemoglobinuria
Cells of the Innate Immune System
There are many types of white blood cells, or leukocytes, that work to defend and protect the human body.
In order to patrol the entire body, leukocytes travel by way of the circulatory system.
The following cells are leukocytes of the innate immune system:
monocytes
monocytes are produced in the bone marrow, circulate for a short period then localise in various tissues
becoming specific macrophages
Functions of Monocytes
The following are some of the functions of monocytes:
1. Monocytes are one of the most important components of the innate immune system as these
differentiate into populations of dendritic cells and macrophages, which are involved in the
regulation of cellular homeostasis.
2. Monocytes also regularly patrol the body for pathogens and regulate an immune response during
infection and inflammation.
3. Monocytes function as phagocytic cells and antigen-presenting cells in the peripheral blood to
remove microorganisms, antigens, and dead or damaged cells.
4. Different subsets of monocytes produce different cytokines that recruit additional cells and proteins
to affected areas to generate an effective immune response.
5. Monocytes are highly plastic and heterogenous as these can change their functional phenotypes
depending on the environmental stimuli.
6. A particular subset of monocytes, called transitional monocytes, is involved in the activation of T
cells.
Macrophages
Macrophages, commonly abbreviated as “Mφ”, are efficient phagocytic cells that can leave the circulatory
system by moving across the walls of capillary vessels. The ability to roam outside of the circulatory system
is important, because it allows macrophages to hunt pathogens with less limits. Macrophages can also
release cytokines in order to signal and recruit other cells to an area with pathogens.

Types of Macrophages
Macrophages can be classified into one of the two opposing phenotypes; classically activated or M1
macrophages and alternatively activated or M2 macrophages.
1. Classically activated macrophages or M1 macrophages
 Macrophages stimulated with a toll-like receptor in the presence of interferon-γ result in the
formation of M1 macrophages.
 These macrophages have an enhanced capacity to present antigen, produce nitric oxide and secrete a
large number of chemokines,
 These macrophages are essential in defense against bacteria which can be damaging to the host as a
result of collateral damage brought about by the defense mechanisms they utilize.
2. Alternatively-activated macrophages or M2 macrophages
 Macrophages activated as a result of exposure to IL-4, IL-3 produced by CD4+ T cells from the
alternatively activated macrophages or M2 macrophages.
 These macrophages are usually produced in the response to parasites and fungi. These express high
amounts of cytosolic arginase and extracellular matrix-related proteins.
 M2 macrophages have the ability to limit inflammation and also play an essential role in tissue repair
and wound healing.
The following are some of the functions of macrophages;
 1. Macrophages are phagocytes involved in the removal of dead cells and cellular debris as a part of
homeostasis. Phagocytosis is one of the principal mechanisms involved in innate immunity.
2. Macrophages also present antigens to other immune cells as a part of initiating an immune response.
These also secrete different chemokines and a wide variety of powerful substances that influence the
activation of cells of adaptive immunity.
3. Macrophages are also involved muscle repair, growth, and regeneration after the inflammation of
different sites.
4. M2 macrophages are also called wound healing macrophages as these limit the extent of the
inflammation and allow tissue repair and regeneration.
5. Since macrophages are scavengers, they continuously remove dead erythrocytes from blood. The
process results in the storage of iron released during the process in the form of ferritin, thus playing a
role in iron homeostasis.
Examples
Macrophages are also of different types depending on the type of tissue they are found on.
1. Alveolar macrophages in the lung.
2. Histiocytes in connective tissues.
3. Kupffer cells in the liver
4. Mesangial cells in the kidney.
5. Microglial cells in the brain.
6. Osteoclasts in the bone.
Mast cells
Mast cells are cells of the immune system of the hematopoietic lineage that remain widely distributed in
vascularized tissues throughout the body.
 Mast cells, unlike other immune cells, do not remain in the blood but remain localized in mucosal
and epithelial tissues throughout the body.
 Mast cells are mononuclear cells consisting of small secretory granules that range in size from 0.2 to
0.8 micrometers. In some cells, the granules are dense enough to obscure the appearance of the
nucleus.
 The cells are oval or irregularly shaped with a single central nucleus. Within the nucleus, densely
packed peripheral chromatin can be observed.
 Most of the cytoplasm is occupied by cytoplasmic granules but few secondary lysosomes. The cells
also have small, finger-like projections from the cell membrane.

Types of Mast Cells

 Mast cells in human beings can be differentiated into two types; mucosal and connective tissue mast
cells.
o The connective tissue mast cells are found primarily in the loose connective tissue and skin,
but these can be found in other connective tissues as well.
o The mucosal type mast cell is found in the gastrointestinal mucosa and peripheral airways.
Functions of Mast Cells
The following are some of the functions of mast cells in the body;
1. Mast cells secrete different pro-angiogenic factors like VEGF, bFGF, TGF-beta, and IL-8. The cells
also release proteases and heparin, which induce permeability of the microvasculature, inducing
angiogenesis.
2. Mast cells are among the first line of defense against antigens entering the body. These cells are thus
important for maintaining the homeostasis of the body, especially the gastrointestinal tract.
 3. In innate immunity, receptors on mast cells bind to antigens, causing a release of inflammatory
mediators like IL-4, TNFα, and IL-6.
4. These cells also help in providing immunity against viral antigens by the release of IFN-α and IFN-β.
5. In adaptive immunity, mast cells process and present antigen via MHCI and MHCII, and these also
activate dendritic cells, which are professional antigen-presenting cells.
6. The most important and prominent function of mast cells is the release of newly synthesized
mediators necessary for the immune response.
7. Mast cells are also involved in an allergic reaction where the over-activation of mast cells can result
in hyper-allergic reactions.
Neutrophils
Neutrophils are a type of white blood cell with multi-lobed nuclei and stainable cytoplasmic granules.
 These are the most abundant granulocytes, occupying about 40-60% of the total number of white
blood cells in the blood.
 Neutrophils are mostly circular ranging in size from 12-15 µm (in humans, the average size is 8 µm).
 Their shape changes into amoeboid once they are activated so that they can extend their pseudopodia
to attack invaders.

Neutrophils Functions
 Neutrophils are the most abundant granulocytes that makeup about 40% of white blood cells and
60% of the immune cells in the blood.
 Neutrophils are the first responders to infection, and they phagocytose bacteria into phagosomes
before hydrolyzing and destroying them.
 These cells also secrete a range of proteins that have antimicrobial effects as well as tissue
remodeling potential.
 Neutrophils have a short lifespan and thus destroy themselves during the degradation of foreign
invaders. New neutrophils are then produced continuously in the bone marrow.
 The neutrophils of another subpopulation, cager neutrophils, perform a transport function of
delivering foreign particles to the target site for the action of killer neutrophils.
Eosinophils
Eosinophils are motile phagocytic cells that play an important homeostatic role in providing defence against
parasitic infections.
 Eosinophils are bone marrow-derived granulocytes that remain in the bloodstream for a shorter
period of time and mostly reside in tissues.
 Eosinophils are granulocytes, measuring in size between 10-16 µm in diameter. The cells contain a
segmented or bi-lobed nucleus where the nuclei: cytoplasm ratio is about 30%.
 An important morphological feature of eosinophils is the content of their cytoplasmic granules,
which contain specific cationic proteins.
 The specific granules of eosinophils are the principal distinctive feature of the cells. These granules
contain a unique crystalloid core, a dense matrix surrounded by a membrane. The core is responsible
for the cardinal properties of eosinophils.
Functions of Eosinophils
The following are some of the functions of eosinophils:
1. The most obvious function of eosinophils is their role in host defense against parasitic infections.
2. Besides, eosinophils are also known to release lipid, peptide, and cytokines that mediate
inflammation and host defense.
3. Some eosinophils also function as antigen-presenting cells as these can be induced to express MHC
class II proteins.
4. Eosinophils are effector cells that release lipid mediators like leukotriene C4, lipoxins, and PAF that
contribute to the acute manifestations of allergic or immunological responses.
5. Eosinophils also have the potential to regulate mast cell functions through the release of granule
proteins and cytokines.
6. A panel of cytokines like IL-2, IL-4, IL-6, IL-12 are released by eosinophils which are capable of
promoting T cell proliferation and activation of Th1 or Th2 polarization.
7. Eosinophils in the lungs has been recognized as a significant contributor to airway hyperreactivity
and asthma.