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Phagocytosis
17/10/2010
Isma’eel
Mohammad & Chemical
Matalqa Mediators
Abu-A’ssi
9 20
v
‫بسم اهلل الرمحن الرحيم‬
Sunday, October 17, 2010
Pathology lecture by Dr.Isma’eel Matalqa
Chapter two; Inflammation
Done by: Mohammad Abu-A’ssi

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Phagocytosis & Chemical Mediators

Some notes about this lecture:
1. Key concepts are written BOLD.
2. Additional notes from the doctor and my own
clarifications are written in italics.
3. Sometimes the doctor was showing a picture or a figure on
the slide, and he was just reading what is written on them,
so these figures are included in this summary.

The doctor declared two announcements before he started

the lecture:
1. Regarding students who did not register for pathology
lab, their names are on the pathology department, and these
students should register before they exceed the allowed number of
absence, otherwise they will be considered failed in the theoretical
subject.
2. Regarding the first exam, it will be on Saturday,
October 30, 2010, and all the lectures before that date are included
in the first exam, which are 6 lectures for cell injury, and other 6
for inflammation. The doctor ensured that each student should be
committed with the information sent to him

PHAGOCYTOSIS

Phagocytosis is the process of ingestion and digestion by

cells of solid substances (other cells, bacteria, microbes, necrotic
tissue or foreign material), so necrotic tissue or foreign material
will be engulfed (phagocytoted), then it will be degraded or
digested be phagocytic cells by several mechanisms.
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There are steps of phagocytosis:
1. Recognition of the foreign material, necrotic tissue,
microbes, etc. by attachment and binding to cellular
receptors. There are cellular receptors at the surface of the
leukocytes which will bind to these foreign bodies which are
usually coated by circulating proteins, they are called
Opsonins such as IgG (immunoglobulin G), C3b (a
complement protein) and collectins. The process of protein
coating to foreign bodies is called Opsoninaization.

Circulating proteins (opsonins) coat the foreign material to

define it as “foreign” and make it recognizable for phagocytic
cells, which then identify it, bind to it by its cellular receptors
and start the process of phagocytosis.

2. Engulfment
3. Fusion of phagocytic vacuoles with lysosomes
4. Killing or degradation of ingested material (iNOS and ROS)

Recognition and attachment by receptors:

Mannose receptors: bind to terminal mannose residues on
microbes cell walls.
They are present at the surface of leukocytes, and they are an
additional mechanism to recognize microbes. (They do not bind to
coating proteins but to specific molecule,i.e. mannose, which is
present only is some microbes and absent in mammalian cells)
Mammalian cells are not recognized by mannose receptors
because they contain terminal sialic acid and N-acetyl
galactosamine. So they are not recognized by these types of
receptors which are present on the leukocytes.

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 Other receptors are:
• Scavenger receptors: oxidized LDL, and microbes.
• Opsonin receptors (high affinity): IgG, C3b,
collectins

Summary for the steps of phagocytosis:

The microbes are first coated with opsonins (which have a high
affinity for binding to microbes), then the leukocytes recognize the
coated microbes and bind to it by opsonin receptors on their
surface, engulfment then occurs, the plasma membrane of the
phagocyte zips up around the microbe. After invagination, the
phagosome (the endocytotic vacuole containing the microbe) will
fuse with lysosomes to form the phagolysosome(which contains
both microbe and lysosomal enzymes). Once the microbe and the
enzymes are together, they stimulate several degradation and
bacteriocidal (bacterial-killing) mechanisms. Most important
mechanisms are the iNOS and ROS. Stimulation of iNOS
(inducible Nitric Oxide Synthase) will lead to the activation of NO
(Nitric Oxide). Stimulation of phagocyte oxidase will lead to the
activation of ROS (Reactive Oxygen Species). These two
mechanisms participate in the killing of microbes in addition to
lysosomal enzymes. In many cases these mechanisms are not really
adequate to kill the bacteria. So these mechanisms are usually
associated with other mechanisms ( we will come on them later) to
completely kill and degrade the microbe.

Bacteriocidal agents: agents that kill the bacteria.

Baceriostatic agents: agents that stop the growth of the
bacteria.

This figure (next page) summarizes the steps:

Note different types of receptors….

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 The generation of the oxygen metabolites is the most
important step in the degradation of microbes. This process is
intiated by a trigger, which is usually an infection, that’s why we
see increase in oxygen metabolites during infections especially
microbial. So during infection, oxygen consumption is increased
because of the increased activity of NADPH oxidase, which
converts O2 to O2- (superoxide ion) and H+;

Under the effect of Dismutase, O2- reacts with H+ to form

hydrogen peroxide H2O2; which is one of the bacteriocidal agents:

If halides are available, like Cl-, which is present within the

cells and in their vicinity, hydrogen peroxide will react with them
forming hydrochlorous radical HOCl-

This reaction is catalyzed be the enzyme Myeloperoxidase

(MPO), this enzyme is the cause of the color of eosinopheric
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granules present in neutrophils. Hydrochlorous radical HOCl- is
the most efficient bacteriocidal agent in the cell. HOCl- is used also
in industry, in manufacturing the chemical compounds of cleaning
and sterilization (Hypex, Flash….)

A summary of the generation of oxygen metabolites:

Ma 3alena,,

How do leukocytes kill infectious agents??

1. Oxygen burst products, which is what we have just discussed
(oxygen metabolites)
2. Bacteriocidal permeability increasing protein, which are
produced by the neutrophils.
3. Lysozyme.
4. Major basic protein, which is a specific protein produced by
the granules within the eosinophils. These kinds of proteins
specifically attach to parasites, and that’s why when you have
a parasitic infection, you will have a larger than normal
number of eosinophils.
5. Defensins.
6. Lactoferrin.

*** In all types of microbial infections we see an increased

number of WBCs generally, but each type of infection is
characterized by a large increase in one of the WBCs, for
example:
Bacterial infection -> large number of neutrophils
Parasitic infection -> large number of eosinophils

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Viral infection -> large number of lymphocytes

Any defect or abnormality in any of these enzymes, receptor

proteins or steps of phagocytosis, might lead to serious diseases.
For example:
1. Patients with Leukocyte Adhesion Deficiency 1 (LAD
1) disease, usually, suffer from recurrent infections in the
lungs, skin, uterine, etc. The cause of this disease is a
defect in CD18 –β subunit of integrin.
2. In the LAD 2 disease, which has the presentation but
more or less in a milder form, the deficiency is in Sialyl-
Lewis X, which is a receptor found on the surface of the
leukocytes.
3. In neutrophil-specific granule deficiency the defect
is absence of specific granules, and we have just
mentioned the importance of neutrophilic granules and
their role in secretion some critical compounds like the
enzyme Myeloperoxidase.
4. Chronic Granulomatous Disease (CGD) has two
types; X-liked CGD and autosomal recessive CGD.
The main cause of both types is a defect in NADPH
oxidase, but in X-liked CGD the defect is in the
membranous NADPH oxidase, while in autosomal
recessive CGD the defect is in the cytoplasmic NADPH
oxidase. The defect in NADPH oxidase leads to decreased
number of oxygen metabolites, but the body partially
overcomes this problem by recruiting more histocytes
(macrophages), so the problem is partially solved. Such
condition of the histocytes will lead to the formation of
“granulomas” (collection of activated macrophages),
and that’s why we call it chronic granulomatous disease.

*** X chromosome is the sexual chromosome that carries

genes (Y chromosome also carries some genes but much less
than X so it is considered to be empty of genes)
*** Autosomal chromosomes are the 22 somatic
chromosomes

5. Myeloperoxidase (MPO) deficiency leads to the

absence of MPO-H2O2 system.
6. Chediak-Higashi disease is caused because of a
deficiency in organelle trafficking. You saw that one of the
steps of phagocytosis is the fusion of lysosomes and
phagosomes, such movement is called organelle
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 trafficking. So if there is a deficiency in organelle
trafficking, the fusion will not occur.

These defects are either inherited or because of a mutation in

a specific gene.

Q) What is the difference between histocytes and macrophages??

A) Monocytes, histocytes and macrophages are all the same, but we
give them different names according to the site of action, if they are
in the blood we call them monocytes, if they are in the tissues we
call them macrophages or histocytes.

In addition to the previous diseases, we have also some

acquired diseases. These diseases are not related to genes, but we
acquire them during our life.
Acquired defects in leukocyte functions:
 Chemotaxis defects
• Burns, diabetes, sepsis, etc.
 Adhesion
• Hemodialysis, diabetes
 Phgocytosis and microbicidal activity
• Leukemia, sepsis, diabetes, malnutrition, etc

CHEMICAL MEDIATORS

There are two sources for chemical mediators; circulating

plasma proteins and cells. There are different types of circulating
plasma proteins which form together different systems, most
important systems are:
 Coagulation / fibrinolytic system (i.e. coagulation cascade)
 Complement system
 Kinins

Cell derived mediators are all elements normally sequestered

in granules, so they are already formed and stored within these
granules which are present in the cytoplasm. The classical example
of these mediators is Vasoactive amines. The other type of cell
derived mediators is the newly synthesized mediators in response
to stimulation, such as PGs (prostaglandins), LT (leukotrienes), O2
species, NO (nitrous oxide), Cytokines and PAF (platelet-activating
factor).( So all these mediators are not formed unless there is
stimulation for their synthesis ). If we have these mediators
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continuously synthesized, this will lead to harmful effects on the
tissue. After these mediators finish their function, some antibodies
will eliminate or inactivate them; otherwise, serious effects to the
tissue might happen.

The following figure is a very important figure which the

doctor ensured the students to study it well and understand it;
otherwise it will be difficult to understand at any stage later.

There are general characteristics of chemical mediators of

inflammation; you need to understand them well to be able to
understand the steps of inflammation later in this chapter. These
general characteristics are:
• They bind to specific cellular receptors, or have enzymatic
activity which can be used to stimulate other cells to
respond. So this is their way of action.
• They may stimulate target cells to release secondary
mediators with similar or opposing functions, for example,
if there is a mediator, e.g. cytokine A, released by
macrophages, it will induce neutrophils to produce cytokine
B which increase and exaggerate anti-inflammatory
reactions, so it has the same function as cytokine A.
Cytokine A also induces neutrophils to produce cytokine C,
which has exactly the opposing function of cytokine A.
• May have limited targets, or wide spread activities
• Short lived function, because of:
 Short half-life (Arachedonic acid metabolites)

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  Inactivated by enzymes (kininases inactivate
bradykinins)
 Eliminated (antioxidants eliminate O2 species –
which are a type of cell derived mediators)
 Inhibited (complement inhibitory proteins)
• If these chemical mediators remain unchecked, unopposed
or activated, they will cause harmful effects. So chemical
mediators have beneficial and harmful effects on tissues,
and the body’s task is to maintain the balance between
beneficial and harmful effects.

Vasoactive Amines, as we said, are the most important

class of cell derived chemical mediators. The main two types of
vasoactive amines are serotonin and histamine. Release of
histamine can be initiated in response to many stimuli, like:
 Physical injury
 Binding of IgE to Fc receptors. In allergy (hypersensitivity),
IgE binding to Fc receptors is stimulated. This, in turn,
induces mast cells to secrete large amounts of histamine. In
treatment of allergy, patients are administered anti-
histamine drugs, which are either histamine receptor
blockers, “mast cell stabilizers” which are sprays (nasal
sprays) that stabilize mast cells, or steroids.
 Anaphylatoxins (C3a, C5a) binding
 Histamine releasing proteins derived from PMNs

***PMNs stands for Polymorphonuclear Neutrophils, another

name for neutrophils.

 Neuropeptides (substance P)
 Cytokines (IL-1, IL-8)

The release of serotonin is mainly activated by:

 Platelets aggregation
 PAF

THAT’S ALL FOLKS

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