Resistance of the Body to Infection 1.

Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

Resistance of the Body to Infection

1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

1. Leukocytes - White Blood Cells, “WBCs”

 Leukocytes (WBCs) are the mobile units of the body’s protective system.
 The importance (value) of WBCs is their transport to areas of serious infection and inflammation,
thus providing a rapid and potent defense against infectious agents.
 Granulocytes and monocytes and B-lymphocytes are formed in the bone marrow.
 Lymphocytes and plasma cells are formed in the lymph tissue.

1.1. General characteristics of Leukocytes

 Count: 4500-11000/µL of blood.
 Presence: after formation, they are transported in the blood to different tissues of the body where
they are needed.
 6 types of WBCs are normally present in the blood:
Granulocytes group:
1. Polymorphonuclear neutrophils [granular cytoplasm, more than 3 lobe nucleus, count 62%]
2. Polymorphonuclear eosinophils [granular cytoplasm, 2 lobe nucleus, count 2.3%]
3. Polymorphonuclear basophils [granular cytoplasm , horse shoe nucleus, count 0.4%]
A granulocyte group:
4. Monocytes [Large size, clear cytoplasm, large acentric nucleus, count 5.3%]
5. Lymphocytes [small size, clear cytoplasm, large round nucleus, count 30%]
6. Plasma cells [present in lymphoid tissues]

Platelets: small cell fragments [count 150,000 – 450,000/µL of blood]

 Phagocytic cells: granulocytes and monocytes engulf and destroy microbes by releasing
antimicrobial or inflammatory agents.
 T-lymphocytes and B-lymphocytes “plasma cells” are parts of the immune system.
 Platelets: play major role in blood clotting (hemostasis)

1.2. Genesis of WBCs

Lymphogenous tissues: the lymph glands, spleen, thymus, tonsils, and pockets of lymphoid
tissue in the bone marrow and Peyer’s patches underneath the epithelium in the gut wall.

1.3. Life Span of WBCs

 Granulocytes in blood: 4-8hours; in the tissue: 4-5 days ; in inflamed tissue: few hours.
 Monocytes: in blood: 10-20hours; in the tissue: mobile tissue macrophage _: months unless
destroyed; fixed tissue macrophage _: years.
 Lymphocytes: total life weeks – months depending on the body’s need.
 Lymphocytes circulate between the blood and lymph.
 Platelets: in blood _ 10 days (10% of platelets replaced per day, average 30,000/day)
1-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

2. Neutrophils and Macrophages Defend Against Infections

 Neutrophils and macrophages attack and destroy invaders (phagocytes).
 Blood neutrophils are mature; blood monocytes are immature; macrophages are mature.
 Mature means: equipped with phagocytic ability, proteolytic enzymes and oxidizing agents (OH-,
H2O2, O2- - all called reactive oxygen species “ROS”).
 Monocytes enter the tissue and begin to swell (increasing their diameters 60-80µm “5folds”, seen by
naked eye) and become macrophage (mature cell)
 Neutrophils and monocytes
 Can squeeze through the pores of the blood capillaries by diapedesis.
 Move through the tissues by ameboid motion (as great as 40 μm/min)
 Attracted to inflamed tissue area by chemotaxis
 Diapedesis: cells send their pseudopods through the pores of the capillaries that are smaller than
their cell size and then cross the membrane into the tissue.
 Chemotactic agents [chemical agents in the tissue that move the cells toward their site] are:
1. Some of the bacterial or viral toxins
2. Degenerative products of the inflamed tissues
3. Several reaction products of the “complement complex” activated in inflamed tissues
4. Several reaction products caused by plasma clotting in the inflamed area, as well as other
substances.
 High concentration of chemotactic agents “concentration gradient” near the source, attracts WBCs
“unidirectional movement”.
 Note:
 Almost no tissue area is more than 50 micrometers away from a capillary.
 Chemotaxis is effective up to 100 micrometers away from an inflamed tissue.

2-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

2.1. Phagocytosis
 Definition: Cellular ingestion (engulfment) of the offending (invading) agent.
 Selectivity: Selective for foreigns but not for normal cells and body structures. It depends on:
1. Most natural tissue structures have smooth surfaces [rough surfaces are phagocytized]
2. Most natural tissue structures have protective protein coats [dead tissues and foreign particles have no
protective coats]
3. Opsonization: Means, immune antibodies adhere to the foreign membranes making them especially
susceptible to phagocytosis.
 Mechanism:
 Antibody binds to foreign antigen.
 The C3 (product of the complement cascade) combine with the antibody.
 The C3 molecule binds their receptors on phagocyte (initiating phagocytosis).
 While the foreign particle on the surface, the
membrane of phagocyte invaginates creating
phagosome.
[Phagosome: is a free-floating phagocytic vesicle
contains the phagocytized particle]
 A single neutrophil can engulf 3-20 bacteria before
becomes inactivated and dies (engulf only
bacteria)
 A single macrophage can engulf 100 bacteria and
can engulf larger particles (RBCs, malarial
parasites)
 Macrophages can extrude the residual digestion
products and survive and function for many more
months. [Macrophages are more powerful phagocytes than neutrophils]

 Digestion mechanism
 Lysosomes and cytoplasmic granules immediately fuse with the phagosome.
 Digestive enzymes and bactericidal agents are dump into the phagocytic vesicle (phagosome + lysosome).
[The phagocytic vesicle now becomes a digestive vesicle and digestion begins immediately]
 Lysosomes of neutrophils and macrophages contain digestive proteolytic enzymes.
 The lysosomes of macrophages (but not of neutrophils) also contain large amounts of lipases, which digest
the thick lipid membranes possessed by some bacteria such as the tuberculosis bacillus.

 Neutrophils and Macrophages Can Kill Bacteria

 Some bacteria have protective coats or other factors that prevent their destruction by digestive enzymes.
 So, bactericidal agents of neutrophils and macrophages can kill most bacteria [even when the lysosomal
enzymes fail to digest them].
 These bactericidal agents are powerful oxidizing (lethal in small amounts) agents [formed by enzymes in the
membrane of lysosomes, phagosomes or peroxisome].
 These are: superoxide (O2−), hydrogen peroxide (H2O2), and hydroxyl ions (OH−), hypochlorite (HOCl)
[Hypochlorite, formed by reaction of H2O2 and chloride ions by the action of lysosomal myeloperoxidase]
 Tubercle bacilli (tuberculosis) resist digestion by lysosomes… why?
 Tubercle bacilli have special resistant coats and secrete substances that resist the killing effects by
neutrophils and macrophages.

3. Monocyte-Macrophage Cell System (Reticuloendothelial System)

 Reticuloendothelial system: Combination of monocytes, mobile macrophages, fixed tissue
macrophages, and a few specialized endothelial cells in the bone marrow, spleen, and lymph nodes.
 Monocyte: Immature WBCs in blood; mobile macrophages: Mature mobile monocyte in tissue; Fixed
tissue macrophages: mature monocytes attached to the tissue and life for months or years.

3-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

 Fixed macrophages:
 Phagocytes, that when appropriately stimulated, can break away from their attachments and
become mobile.
 Respond to chemotaxis and all the other stimuli related to the inflammatory process.
 Thus, all tissue areas have “monocyte-macrophage system”.
[The term reticuloendothelial system = monocyte-macrophage system]

Fixed tissue macrophages

1. Histiocytes: Tissue Macrophages in the skin and subcutaneous tissues.
2. Macrophages in the lymph nodes: Entrapped by the reticular meshwork of lymph nodes
3. Alveolar macrophages in the lungs
 They digest the digestible particles and release the digestive products into the lymph
 Indigestible particles are encapsulated by macrophages that form a “giant cell” capsule around the
particle until it can be slowly dissolved [frequently formed around tuberculosis bacilli, silica dust
particles, and carbon particles].
4. Kupffer Cells: Macrophages in the liver sinusoids
 Kupffer cells prevent (none of the) bacteria pass into the general systemic circulation.
 Kupffer cells phagocytized single bacterium in less than 0.01 second.
5. Macrophages of the spleen and bone marrow
 These macrophages entrapped by the reticular meshwork of the spleen and bone marrow.
[The spleen is similar to the lymph nodes, except that blood, instead of lymph, flows through the tissue
spaces of the spleen]

4. Inflammation: Role of Neutrophils and Macrophages

4.1. Inflammation
 Tissue can injure by: physicals (trauma, extreme heat and cold, electricity, radiation), chemicals
(Metals, solids, liquids or gases), and biological agents (bacteria, parasite and viruses).
 Injured tissue releases inflammatory mediators:
(1) Histamine (released by mast cells)
(2) Bradykinin (released by damaged membranes)
(3) Serotonin (released by platelets)
(4) Prostaglandins (released by damaged membranes)
(5) Complement system reaction products (in the blood that enter the tissue)
(6) Reaction products of the blood clotting system
(7) Lymphokines (released by sensitized T-lymphocytes)
 Injured and surrounding uninjured tissues undergo complex histological changes called inflammation.
 Inflammation is characterized by:
Vascular changes:
 Vasodilation of the local blood vessels (excess local blood flow)
 Swelling of the tissue cells
 Increased capillary permeability -leakage “exudation” of large quantities of fluid into the interstitial
spaces
 Clotting of the fluid in the interstitial spaces [due to leakage of fibrinogen and other proteins from the
capillaries]
Cellular changes
 Migration of large numbers of granulocytes and monocytes into the tissue (infiltration).
 Several of these inflammatory mediators strongly activate the macrophage system
 Within a few hours, the macrophages begin to devour “engulf” the destroyed tissues
 At times, however, the macrophages also further injure the still-living tissue cells.

4-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

“Walling-Off” Effect of Inflammation

 One of the first results of inflammation is to “wall-off” the area of injury from the remaining tissues.
 Tissue spaces and the lymphatics in the inflamed area are blocked by fibrinogen clots
 Thus, after a while, fluid barely flows through the spaces.
 This walling-off process delays the spread of bacteria or toxic products.
 The intensity of the inflammatory process is usually proportional to the degree of tissue injury.
[The walling-off process develops rapidly or slowly according to the intensity of tissue damage]
 Example: develop rapidly in Staphylococcal infection. Why?
 Staphylococci invade tissues and release extremely lethal cellular toxins (high tissue damage).
 Inflammation develops rapidly (rapidly than staphylococcal multiplication and spread).
 Rapid walled off staphylococcal infection prevent local infection from spreading through the body.
 Example: develop slowly in Streptococcal infection. Why?
 Streptococci do not cause such intense local tissue destruction.
 The walling-off process develops slowly over many hours, while many streptococci reproduce and
migrate.
 As a result, streptococci often have a far greater tendency to spread through the body and cause
death than do staphylococci, even though staphylococci are far more destructive to the tissues.

4.2. Macrophage and Neutrophil Responses during Inflammation

First line of defense against infection: Tissue Macrophages.
 Infectious agent evokes inflammatory response.
 Products of inflammation activate mobile macrophages to immediately begin phagocytic actions
 Next, fixed tissue macrophages break loose from their attachments and become mobile during the first hour
or so.
 The numbers of these early mobilized macrophages often are not great, but they are lifesaving.

Second line of defense against infection: Neutrophils.

 Within the 1st hour from beginning of inflammation, inflammatory cytokines and others attract neutrophils.
 Large numbers of neutrophils begin to invade the inflamed area from the blood.
 Neutrophils are mature cells they begin their scavenger functions of killing bacteria and removing foreign
matter
 Inflammatory cytokines - tumor necrosis factor “TNFα” and interleukin-1 “IL-1” causes:
1. Margination
 Increased expression of adhesion molecules on the surface of endothelial cells in the capillaries and
venules.
[Adhesion molecules = selectins and intercellular adhesion molecule–1 (ICAM-1)]
 Adhesion molecules react with complementary integrin molecules on the neutrophils
[The neutrophils stick to the capillary and venule walls in the inflamed area]
 This enhances diapedesis
[By loosen the intercellular attachments between the endothelial cells of the capillaries and small venules]
[These allowing openings large enough for neutrophils to crawl directly from the blood into the tissue
spaces]
2. Chemotaxis
 Neutrophils directed to the injured tissues.
 Thus, within several hours after tissue damage begins, the area becomes well supplied with neutrophils.

Acute Increase in the Number of Neutrophils in the Blood—“Neutrophilia”

 Neutrophilia: increased blood neutrophil count.
 Within a few hours after the onset of acute, severe inflammation, blood neutrophils count sometimes
increases 4-fold to 5-fold (15,000 to 25,000/µL) ___ differential count >90%

5-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

 Mechanism: Products of inflammation enter the blood stream  transported to the bone marrow 
mobilize stored bone marrow neutrophils into the blood.
 Neutrophilia makes more neutrophils available to the inflamed tissue area.

Third line of defense against infection: Second macrophage invasion

 Monocytes from the blood enter the inflamed tissue and after 8 hours enlarge to become
macrophages.
 The storage pool of monocytes in the bone marrow is much less than that of neutrophils.
 Thus, the buildup of macrophages in the inflamed tissue area is slower than of neutrophils (requiring
several days to become effective).
 After several days to several weeks, the macrophages finally come to dominate the phagocytic cells of
the inflamed area (because of greatly increased bone marrow production of new monocytes)
 Macrophages can phagocytize far more bacteria (about 5-times than neutrophils) and larger particles,
including even neutrophils and large quantities of necrotic tissue (i.e. clear the inflamed tissue)
 Also, the macrophages play an important role in initiating the development of antibodies.

Fourth Line of Defense against infection: Increased production of granulocytes and monocytes
by the bone marrow
 Stimulus from inflamed tissue stimulates progenitor cells of the marrow.
 These cells are committed to form granulocytes and monocytes.
 The process requires 3 to 4 days for these cells to leave the bone marrow.
 As long as the stimulus continues, the bone marrow continues production of these cells.
 The production rate can continue for months and even years.
 The production rate can be 20-50 times the normal.

4.3. Feedback Control of the Macrophage and Neutrophil Responses

 5 factors have dominant roles in feedback control of the macrophage response to inflammation:

 Factors in 3, 4, and 5 are the main cause of increased bone marrow production of granulocytes and
monocytes.
 GM-CSF stimulates both granulocyte and monocyte production.
 G-CSF and M-CSF stimulate granulocyte and monocyte production, respectively.
 The feedback: Stimulus  inflammation  activate cells  bone marrow response production of
defensive WBCs  help remove the cause of the inflammation  remove the stimulus  stop bone
marrow production

6-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

7-10
 Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

4.4. Formation of Pus

 After engulfment of large numbers of bacteria and necrotic tissue:
 All the neutrophils eventually die
 Many, if not most, of the macrophages eventually die.
 Several days after inflammation begin, a cavity is often excavated in the inflamed tissues containing:
 Varying portions of (a) necrotic tissue (b) dead neutrophils and dead macrophages (c) tissue fluid
 This mixture is of (a), (b) and (c) commonly known as pus
 Several days after suppressing the infection
 The dead cells and necrotic tissue are autolyze.
 The end products are eventually absorbed into the surrounding tissues and lymph
 Most of the evidence of tissue damage is gone

5. Eosinophils
 Count: 2% of all the blood leukocytes.
 Phagocytic power: weak
 Chemotaxis: yes
 Cytoplasmic granules: are modified lysosomes, contain hydrolytic enzymes and histaminase
 Their blood count increase (eosinophilia):

1. Parasitic infection
 Eosinophils preventing excess spread of the parasitic infection
Mechanism:
a) Attach to parasitic surface (in larval stage) through special surface molecules
b) Release killing substances (hydrolytic enzymes, ROS and larvacidal polypeptide)
[ROS= lethal reactive oxygen species; larvacidal polypeptide also called major basic protein]
Why they migrate there?
 Tissue parasitic antigenicity causes the release of chemotactic agents
 Eosinophils migrate from blood into tissues diseased by parasites
 Note: Most parasites are too large to be phagocytized by eosinophils or other phagocytes.
 Example of parasitic diseases
 Schistosomiasis: invade any body’s tissue (in many developing countries of Asia, Africa, and South
America)
 Trichinosis: Invade body’s muscles by the Trichinella parasite (“pork worm” – eating undercooked
infested pork)

2. Allergic reactions
 Eosinophils preventing excess spread of the local inflammatory process by:
a) Detoxifying histamine released by the mast cells and basophils
b) Probably also phagocytize and destroy allergen-antibody complexes
Mechanism:
 Eosinophils migrate from blood into tissues showed allergic reactions such as:
 Peribronchial tissues of the lungs in people with asthma
 The skin after allergic skin reactions

Why they migrate there?

 Tissue mast cells and blood basophils participate in allergic reactions:
 They release histamine in tissue and blood (local vasodilation; sensory nerve sensitization)
 They release eosinophil chemotactic factor (eosinophils to migrate toward the inflamed allergic tissue)

8-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

6. Basophils
 Count: 0.4% of all the blood leukocytes.
 Phagocytic power: weak
 Chemotaxis: yes
 Blood basophils are similar to tissue mast cells
 Basophils and mast cells cytoplasmic granules contain
(a) Heparin (b) histamine (3) bradykinin (4) serotonin (5) slow-reacting substance of anaphylaxis
(6) number of lysosomal enzymes
 Both cells liberate heparin into the blood – prevent blood clotting or coagulation.
 Both cells liberate histamine into the blood – local vasodilation
 Basophil blood count increase (basophilia) during allergic reactions:
Mechanism
 Chemotactic agents include:
(a) Lymphokines “from activated lymphocytes” (b) Complement fragments “C5a” (c) Leukotrienes
“LTB4”
 The antibodies immunoglobulin E (IgE) that causes allergic reactions attached to mast cells and
basophils
 Antigen-antibody reaction causes cell rupture and release of granular substances
 These substances cause local vascular and tissue reactions that cause many, if not most, of the allergic
manifestations.

7. Leukopenia (overview)
 Definition: Low blood count of WBCs (low bone marrow production)
 Consequences: leaves the body unprotected against many bacteria and others invaders.
 Significance:
 All the mucous membranes of the body are constantly exposed to large numbers of bacteria.
[Mucous membranes: mouth, entire GIT especially the colon, vagina, urethra, eye, respiratory tract]
[Bacteria like: various spirochetal, pneumococcal, streptococcal bacteria, E. coli]
 Any decrease in the number of WBCs immediately allows invasion of adjacent tissues by bacteria
 Within 2 days after the bone marrow stops producing WBCs “leukopenia”:
 Mucus membrane ulceration
 Severe respiratory infection might develop
 Bacteremia: Bacteria from the ulcers rapidly invade surrounding tissues and the blood
 Septicemia: Bacterial toxins released into the blood
 Without treatment, death often ensues in less than a week after acute total leukopenia begins.

 Pathophysiology: bone marrow depression “bone marrow aplasia”

 Causes:
 Irradiation of the body by x-rays or gamma rays
 Exposure to drugs and chemicals that contain benzene or anthracene nuclei
 Chloramphenicol (an antibiotic)
 Thiouracil (used to treat thyrotoxicosis)
 Various barbiturate hypnotics on rare occasions
 Prognosis:
 After moderate injury and removal of the cause  recovery
 Undestroyed myeloblasts, and hemocytoblasts are capable of regenerating the bone marrow
 Treatment:
 Transfusions + antibiotics and other drugs to ward off infection
 Within weeks to months the new bone marrow return normal blood cell concentrations

9-10
Resistance of the Body to Infection 1. Leukocytes, Granulocytes, the Monocyte-Macrophage System, and Inflammation

8. Leukemias
 Definition: Uncontrolled production of WBCs
 Blood picture: greatly increased numbers of abnormal WBCs in the circulating blood
 Pathophysiology: cancerous mutation of a myelogenous or lymphogenous cell.
 Types: Lymphocytic and Myelogenous leukemia

1. The lymphocytic leukemia

 Caused by cancerous production of lymphoid cells
 Usually beginning in a lymph node or other lymphocytic tissue
 Spreads to other areas of the body

2. Myelogenous leukemia
 Cancerous production of young (partially differentiated) myelogenous cells
 Begins in the bone marrow
 Spreads throughout the body
 WBCs are produced in many extramedullary tissues—especially in the lymph nodes, spleen, and liver
 Subtypes: neutrophilic leukemia, eosinophilic leukemia, basophilic leukemia, or monocytic leukemia.

 Features:
 Leukemia cells are bizarre (unusual)
 Undifferentiated
 Not identical to any of the normal WBCs
 Nonfunctional “especially the very undifferentiated cells”
 Prognosis
 Acute leukemia (lethal):
The cells are undifferentiated
Leading to death within a few months if untreated
 Chronic leukemia:
 The cells are more differentiated
 Develop slowly over 10-20 years

8.1. Effects of Leukemia on the Body

1. Metastasis
 Growth in abnormal areas of the body
2. Bone pain and eventually bone fracture
 Invade the surrounding bones
3. Development of infection
 All leukemias eventually spread to the spleen, lymph nodes, liver, and other vascular regions
4. Severe anemia and thrombocytopenia “bleeding tendency”
 Lack of the normal RBC and platelets production
5. Cachexia:
 Loss of body weight and muscle mass, and weakness (energy depletion)
 Caused by excessive use of metabolic substrates by the growing cancerous cells.
 Deficiency of specific amino acids and vitamins due to over consumption by rapidly reproduced leukemic
cells
 Excessive utilization of amino acids by the leukemic cells causes breakdown of muscle fibers.
 Thus, while the leukemic tissues grow, other tissues become debilitated.
 After metabolic starvation has continued long enough, this factor alone is sufficient to cause death.

10-10