Leucocytes (White blood cells)

1. Granulocytes or polymorphonuclear leucocytes (polymorphs) – granules in cytoplasm

a. neutrophils
b. eosinophils
c. basophils

2. Non- granulocytes (mononuclear leucocytes) – without granules in cytoplasm

a. lymphocytes
b. monocytes

Neutrophils
Morphology
• Size: 10 – 12 m in diameter
• Nucleus: 1- 4 or > lobes
• Granules: 0.02-0.5 m in diameter
purplish (take up both acidic and basic stains)

Functions of Neutrophils

Neutrophils have amoeboid movement (most active among WBCs) and strong phagocytic power.
The neutrophils seek out, ingest and kill bacteria and are called the body's first line of defence against bacterial
infections.
When bacteria enter the body increased production of neutrophils from bone marrow and inflammatory response
occur.
Neutrophils leave the circulation by diapedesis.
C5a, leukotriene and polypeptides from lymphocytes, mast cells and basophils attract neutrophils to the infected
area (chemotaxis).
IgG and complement proteins facilitate phagocytosis (opsonization).

Killing mechanisms

a. Endocytosis (phagocytosis)
Bacteria bind to receptor in neutrophil cell membrane triggering endocytosis (phagocytosis).

b. Respiratory burst
A sharp increase in O2 uptake (respiratory burst) and activation of NADPH oxidase occur. This produces toxic
oxygen metabolites (superoxide anions, O2-).

NADPH + H+ + 2O2→ NADP + 2H+ + 2O2-

2 O2- + 2H+ SOD

H2O2 + O2

H2O2 and O2- are oxidants and effective bactericidal agents.

c. Exocytosis (degranulation)
By exocytosis, lysosomes discharge their contents into phagocytic vacuole and into extracellular medium
(degranulation).These contents contain proteases that break down bacterial components, defensins and
myeloperoxidase. Myeloperoxidase convert Cl-, Br-, I- and SCN- to hypothalous acid (eg. HOCl, potent oxidant).
Toxic oxygen metabolites and HOCl produce an effective killing zone around neutrophils and inside the phagocytic
vacuole.

Neutrophils release thromboxanes, leukotrienes and prostaglandins for inflammatory process.

Eosinophils
Morphology
Size: 10- 12 µm in diameter
Nucleus: Bilobed
Granules:
• Large, coarse (0.7 – 1.3 µm in diameter)
• Take-up acidic stain (red)
• Lysosomal enzymes
• Plasminogen
• Histamine

Functions of Eosinophils

Properties : they show amoeboid movement but phagocytic activity less than neutrophils.

Function :

1. Eosinophils attack parasites. Eosinophil granules contain major basic protein (MBP) that damage the larva stage
of parasites.
2. Eosinophils produce leukotriene C4 and PAF (platelet activating factor) that are involved in allergic reaction.
Some products inactivate the mediaters released from mast cell.

The circulating eosinophil level is often elevated in allergic diseases (e.g. hay fever, bronchial asthma, food allergy),
skin diseases and worm infestation.

Basophils
Morphology
Size: 8 - 10 µm in diameter
Nucleus: multi-lobed
Granules:
• Large, coarse (1 µm in diameter)
• Take-up basic stain (blue)
• 5-HT (Serotonin)
• Histamine
• Heparin
Function:
Basophils release histamine and other inflammatory mediators when activated by histamine releasing factor
secreted by T lymphocytes. They are essential for immediate – type of hypersensitivity reactions (e.g. severe
anaphylactic shock).

Monocyte
Morphology
Diameter: 16-22 m (the largest cell of WBC)
Nucleus: Single large nucleus, Horse-shoe shaped,Eccentrically placed
Cytoplasm: Clear bluish cytoplasm
Properties:Amoeboid movement (phagocytic power)
Functions: Monocytes functions as macrophages

The macrophage system

Macrophages are mobile cells that are capable of wandering through tissues and engulfing micro-organism, effete
erythrocytes and tissue debris. (Scavenger function)
Macrophages originate from bone marrow. Monocytes enter the circulation from bone marrow and circulate for
about 72 hours. Then enter the tissue and are transformed into tissue macrophages. In the tissue, they may persist
for about 3 months.

Components of Macrophage system

1. Blood – monocytes
2. Bone – osteoclasts
3. Brain – microglia
4. Liver – Kuffer cell
5. Lungs – alveolar macrophages
6. Lymph nodes, bone marrow and spleen – reticulum cells
7. Connective tissues – histiocytes

Function of macrophages or monocytes

1. Phagocytosis and digestion
a. Defence : ingestion and killing of microorganisms
b. Scavenger
: disposal of old, damage or dying cells. e.g. aged RBCs, polymorphonuclear leucocytes, tissue debris and firbrin
: inorganic particulate material such as carbon and dust particle
: organic material such as thorns and spicules
c. Antigen-processing and presentation to T and B lymphocytes.
2. Secretory function: secretion of cytokines and up to 100 different substances.
IL-1 – support lymphocyte production and antibody production
CSF – regulate haemopoiesis
PGE
Clot promoting facotors
Lymphocytes
Morphologically
1. Large Lymphocyte
Size: 10 µm in diameter
Nucleus: Large
Cytoplasm: plenty
2. Small Lymphocyte
Size: 6 - 8 µm in diameter
Nucleus: Rounded
Cytoplasm: scanty
Functionally
B lymphocytes, T lymphocytes (80%) and non B non T lymphocytes (the natural killer cells)
Functions:
Lymphocytes are key constituents of the immune system.
NK cells are involved in natural or innate immunity.
B and T lymphocytes are involved in acquired or adaptive immunity.

Role of Lymphocytes in Immune System

Lymphocytes are key constituents of the immune system.
Morphologically lymphocytes can be divided into small and large lymphocytes.
Functionally, they can be classified into B lymphocytes, T lymphocytes and non-T non-B lymphocytes (Natural
killer cells).
The majority of lymphocytes in the circulation are T lymphocytes (80%).

T cells differentiate into

1. Helper T cells or Inducer T cells
2. Cytotoxic T cells
3. Suppressor T cells
4. Memory T cells
B cells differentiate into Plasma cells and memory B cells.

There are two types of immunity.

1. Natural or innate immunity
2. Acquired or adaptive immunity which involves humoral immunity and cellular immunity.

NK cells are involved in natural or innate immunity.

B and T lymphocytes are involved in acquired or adaptive immunity.

NK cells are involved in Natural or Innate Immunity. It is first line of defense against infections. It also attacks
tumours and foreign cells. The receptors in NK-lymphocytes recognize bacteria. They exert their effects by way of
complement and other systems and by osmosis lysis or apoptosis (programmed cell death). NK cells secrete
cytokines and activate the acquired immune response.
B lymphocytes and T lymphocytes are involved in Acquired or adaptive immunity. The key to acquired immune
system is a remarkable ability of lymphocytes to produce antibodies (for B cells) and cell surface receptors (for T
cells). The immunity system remembers and produces more rapid and greater response on second exposure.

B lymphocytes are involved in Humoral Immunity. They produce circulating antibodies (Immunoglobulin). There
are 5 types of immunoglobulin (IgG, IgA, IgM, IgD, IgE). It is the major defense against bacterial infection.
These antibodies protect their host by
• binding to and neutralizing some protein toxins
• opsonizing bacteria (facilitating phagocytosis)
• fixing and activating complement (helping cell lysis)
• blocking attachment of some virus to cells
• activating NK cells

T lymphocytes are involved in cellular Immunity. It is responsible for delayed allergic reaction, rejection of
transplant tissues, defense against tumours and major defense against infection due to viruses, fungi and a few
bacteria such as tubercal bacillus.
Helper T cell I promotes differentiation of T lymphocytes into cytotoxic T cells.
Helper T cell II promotes differentiation of B lymphocytes into plasma cells.
Cytotoxic T cells destroy transplanted and other foreign cells. They kill by inserting perforins and by initiating
apoptosis.
Suppressor T cells immunoregulate by suppression of both T and B lymphocytes.
Memory B and T cells do not make initial response but are readily converted to effector cells (plasma cells and
cytotoxic T cells) by a later encounter with the same antigen. Unlike other lymphocytes, they persist in the body
without dividing for months or even years.
Completement system

• A system of plasma proteins (enzymes)

• Activated by immune responses, contact with microorganisms & tumor cells
• Mediate innate as well as acquired immune responses
• Complement the effects of antibodies
• Opsonization, Chemotaxis & Lysis of cells by perforins

Recognition of self
T and B cells do not form antibodies against the cells of self (recognize the self).
Reasons
1. Clonal deletion – during embryo development, eliminate the self antigens in the thymus.
2. Clonal anergy – prolonged hyporesponsive state of activated B and T cells on high concentrations of antigen in
the absence of lymphokines.
2. Inhibited by suppressor T cells
Failure to eliminate antiself antibodies → Autoimmune diseases
eg. diabetes mellitus, myasthenia gravis, multiple sclerosis, hyperthyroidism (Grave’s disease)

Cytokines
• Hormone-like chemical messengers
• Secreted by lymphocytes, macrophages and many other cells
• Act generally in a paracrine fashion
• Regulate immune responses, stimulate haemopoiesis, increased circulating neutrophils and their
adhesion to vessel wall, increased capillary permeability, hepatic protein synthesis and synthesis and
secretion of CRH and cause fever
• Once the amino acid sequence is known, its name is changed to interleukin.
• Cytokines that cause chemotaxis are called chemokines.

Acquired Immune Deficiency Syndrome

• HIV virus, retro virus
• decrease in Helper T lymphocytes (CD4)
• failure of proliferation of T and B lymphocytes

Leucopoiesis (Formation of leucocytes)

Sites
• Granulocytes: Bone marrow
• Lymphocytes: Bone marrow (adults), Yolk sac, liver, spleen (fetus)
• Monocytes: Bone marrow

They are formed extravascularly and enter the circulation by diapedesis.

75% of cells in the bone marrow – leucopoiesis (shorter lifespan)
25% of cells in the bone marrow – erythropoiesis (longer lifespan)
Total WBC = 4 000 – 11 000 cells/μL
Differential %
- Neutrophil (50 - 70%)
- Lymphocyte (20 - 40%)
- Monocyte (2 - 8%)
- Eosinophil (1 - 4%)
- Basophil (0 - 0.4%)

Variations in WBC count

Type of Cell > Normal < Normal
Total WBC Leucocytosis Leucopenia
Lymphocyte Lymphocytosis Lymphocytopenia
Monocyte Monocytosis Monocytopenia

Factors controlling leucopoiesis – CSF (colony stimulating factors)

Physiological Variations
Increased in:
1. Pregnancy
2. Parturition
3. Newborn
4. After meal
5. Afternoon tide – diurnal variation
6. After exercise

Pathological Variations
1. Neutrophilia → Acute infections, bleeding, burns, bone fracture and trauma
2. Eosinophilia → Parasitic infestations, asthma and allergic diseases, skin and parasitic infestations
(ascariasis, filariasis)
3. Lymphocytosis → Chronic infections, typhoid, TB, syphilis
4. Monocytosis → Malaria, typhoid and paratyphoid
5. Basophilia → Myeloproliferative disorders and anaphylatic shock
6. Leucopenia → Bone marrow depression, chronic malnutrition and starvation
7. Steroids effect → ↑neutrophil count, ↓lymphocyte count and eosinophil

Lifespan of WBCs
Variable
• Neutrophils → 2 days to 2 weeks
• Monocytes → 3 days in circulation
• Macrophages → 3 months in tissues
• Lymphocytes → Several years
• Aged leucocytes → Destroyed in the liver, spleen, bone marrow and lymph nodes by Macrophages
• In the battle field → Many leucocytes die + Necrotic tissue → Pus