WHITE BLOOD CORPUSCLES (WBCs)

OR
LEUCOCYTES

- Dr Kiran Thorat
• Leucocytes / WBCs perform defense functions of the body

• There are two series of leucocytes:

1. Myeloid Series
2. Lymphoid Series
• Each cell line has separate pathway of development from distinct
primitive cells
• Classification – 2 types -

A) Granulocytes &
B) Agranulocytes
A) Granulocytes

1) Neutrophils - 50 – 70% 3000 – 6000/cmm

2) Eosinophils - 1 – 4 % 150 – 300/cmm

3) Basophils - 0 – 1% 10 – 100/cmm
B) Agranulocytes

1) Lymphocytes - 20 – 40% 1500 – 2700 / cmm

2) Monocytes - 2 – 8% 300 – 600 / cmm

• Leucocytes have three phases in their life:
1. Marrow Phase
2. Circulatory Phase
3. Tissue Phase
• Marrow phase - WBCs develop from progenitor cells where they proliferate
& grow into mature leucocytes & get released into
circulation

• Circulatory phase - WBCs circulate for few hours before entering into
tissues where they live a longer period of their life
• Tissue phase - WBCs enter the tissues where, monocytes become
macrophages

- Usually, life span of WBCs in –

- marrow phase - 6 to 10 days

- circulatory phase - 6 to 8 hours &

- tissue phase - 4 to 5 days

• Leucopoiesis – production maturation & development of leucocytes
• Regulation of Leucopoiesis –

- mainly regulated by cytokines, especially CSFs, ILs & TNFs

- Also by hemopoietic growth factors (HGFs) – relesed by T lymphocytes,

monocytes, fibroblasts, endothelial cells, liver & kidney

• CSFs that influence leucopoiesis are GM-CSF, G-CSF & M-CSF -

- secreted by fibroblasts, vascular endothelial cells, monocytes, T

lymphocytes & macrophages

- (+) differentiation & proliferation of PHSCs into committed cells

- also helps in differentiation of CFU into granulocyte & monocyte

precursors & their further development into mature cells
• Interleukins -

• IL-1, IL-6 & IL-3 - promote maturation of stem cells

• IL-5 / eosinophilic growth factor - helps in development of eosinophil

• IL-3, IL-4 - facilitate development of basophils

• IL-2 - inhibits myelopoiesis

• Tumor Necrosis Factor (TNF) –

- One of the proteins capable of inducing necrosis of tumor cells

- produced mailny by macrophages & also by lymphocytes, NK cells,

neutrophils, mast cells, eosinophils & neurons

- helps in proliferation & differentiation of stem cells

• TLC - At birth - 20,000/cmm of blood
Adults - 4000 - 11000/cmm

• Leucopenia – starvation, typhoid, viral & protozoal infections, BM

depression

• Leukocytosis – newborn, evening (min. in the morning), exercise, inj of E or

NE, stress, Pregnancy, menstruation & lactation,
administration of steroids, any pyogenic or pyrogenic
infection
• Leukemia -
• Physiological leukocytosis –

- ↑ in number of circulating leucocytes in response to physical exercise or to

↑ ed levels of adrenalin in healthy humans

- mechanism underlying this phenomenon & its physiological implications

are not known
- adrenalin is the most important mediator in regulation of leukocytosis

- Before entering the circulating blood leucocytes are stored intra- &/or extra-
vascularly in several organs like lungs, spleen, lymphatic tissue &
haematopoietic BM
• Neutrophils -

• Morphology - 10 –14 μm

Nucleus - purple, multilobed,

3 lobed nucleus functionally mature
• Cytoplasm –

- slight blue & granular

- granules (acidophilic & basophilic), proteins &

lipids

- glycosidases, sulphatases, phosphatases ,

proteolytic enzymes

- NADPH oxidase, Sodium dismutase (SOD),

myeloperoxidase
• Phagocytic Properties
of Neutrophils/WBCs -

1) Margination
2) Diapedesis
3) Chemotaxis
4) Amoeboid motion
5) Opsonization
6) Phagocytosis
7) Degranulation
• Phagocytic properties of Neutrophils -

Respiratory burst (sharp ↑in O2 consumption

by neutrophil during phagocytosis for
production of ROS)
↓
activation of NADPH Oxidase to form
superoxide
↓
superoxide is converted in to H2O2 by SOD
↓
H2O2 is used by myeloperoxidase to oxidize
chloride ions into HOCl
↓
H2O2 & HOCL mediate bactericidal activity
• Physiological basis of the use of H2O2 & HOCL in clinical practice –

1. Washing of wounds with H2O2 while dressing

2. Microbiological slides are dipped Na-hypochlorate solution after being

used

3. IV infusion of dilute HOCl was actually used for treatment of bacterial

sepsis in battlefield hospitals during World War I, before the advent of
penicillin & other antibiotics
• Functions of Neutrophils

1) Phagocytosis

2) Production of endogenous pyrogens

 • Disorders of Phagocytic functions

1) Agranulocytosis / Granulopenia 5) Congenital Neutropenia

2) Myeloid leukemia 6) Congenital myeloperoxidase

deficiency
3) Neutrophil hypomotility

4) Chronic granulomatous disease

of childhood
• Neutrophilia -

• Physiological – exercise, inj. Epnephrine, pregnancy,menstruation

• Pathological - acute infections, tissue destruction,burns,hemorrhage, MI

• Neutropenia - children, viral infection, typhoid/paratyphoid fever, BM

depression
• Arneth count & its clinical significance –

• Age of Neutrophil α number of lobes of its nucleus & is named for Josef
Arneth

N1 – 5% N4 – 18%
N2 – 30% N5 & N6 – 2%
N3 – 45%
• Clinical significance of Arneth count –

- It reflects the state of BM activity

• N1+N2+N3 ˃ 80% - Regenerative shift – pyogenic infections, hemorrhage,

BM hyperplasia

• N4+N5+N6 ˃ 20% - Degenerative shift – Pernicious anemia, Vitamin

deficiencies, BM aplasia
• Eosinophils - 10 – 14 μm

Nucleus - purple in color,

spectacle shaped

Cytoplasm - acidophlic, light pink,

granules coarse,
brick red in colour
• Functions -

1. Antiallergic – E collect at allergy site & release enzymes like histaminase

to limit the effects of inflammatory mediators like histamine

2. Antiparasitic – E attack parasites that are too large to be engulfed by

phagocytosis → granules secrete chemicals like MBP,
ECP, EPO, EDN → toxic to larvae of parasites
 Eosinophilia Eosinopenia

1. Helminthic infections 1. Glucocorticoid therapy

2. Allergic diseases 2. Cushing’s syndrome
3. Drug reactions (drug allergy) 3. Aplastic anemia
4. Eosinophilic leukemia 4. Drug-induced
5. Tropical pulmonary eosinophilia agranulocytosis
6. Addison’s disease
7. Eosinophilia-myalgia syndrome
• Basophils - 10 – 14 μm, granules coarser &
less in number, blue & present
in cytoplasm & over the nucleus

- Nucleus kidney or S shaped

• Functions -
1) Secrete histamine & ECF-A

2) Secrete heparin
• Basophilia - chickenpox, smallpox, TB, influenza

• Basopenia - gluco-corticoids, drug induced

• Mast cells - are round or elongated with non
segmented nucleus

- Granules are numerous, smaller in size

- secrete histamine, heparin, chondroitin

sulphate, carboxypeptidase, cathepsin, ECF-A
& neural protease

- Two types - mucosal mast cells (mast cell

present in the mucosa) &
connective tissue mast cell
- Basophils & mast cells are mainly involved in allergic reactions

- Mediators such as histamine & ECF-A released by degranulation produce

antimicrobial & anti-host effects

- usual stimulus for basophil & mast cell degranulation is an allergen

SCF - Stem Cell Factor
• Lymphocytes – Classification –

A) Morphological – Large & Small

• Large - 10 – 14μm, large & centrally placed

nucleus, clear & thin rim of blue
cytoplasm

• Small - 8 - 9 μm, nucleus circular &

eccentrically placed, cytoplasm clear
blue
B) Functional classification –

- Effector cells primarily involved in adaptive / acquired immune response

are the T & B lymphocytes

- Total lymphocytes count in blood = 1500 – 2700 / cmm

= 1.3 – 4.0 × 109 / Liter

T cells – 50 – 70 % &
B cells – 10 - 20 %
• ‘Natural killer’ (NK) cells – A third population of lymphocytes

- so-called because being able to kill neoplastic or virally infected cells

without prior exposure or sensitization

- are atypical lymphocytes & generally considered to be part of the innate

response
• Lymphocytosis - children, chronic infections,
lymphatic leukaemia, viral infections

• Lymphocytopenia - hypoplastic BM, AIDS

• Monocyte - 10 -18 μm, irregular shaped,
nucleus round or kidney
shaped, eccentric

- cytoplasm clear , pale blue

- occasionally contain fine purple dust

like granules (Azur granules) & gives it
ground glass appearance
• Functions –

1. Phagocytosis & microbial killing - kills intracellular pathogens like bacteria

viruses & parasites by active phagocytosis

• Monocyte is the second line of defense against infections

2. Release of cytokines – that carry out different physiological functions –

• IL-1 & IL-6 - essential for coactivation of immunological responses

• TNF-α & interferons - facilitate killing of viruses & other microbial

organisms

• Complement factors - augment the local tissue defense responses

• Growth factors like -

- GM-CSF, M-CSF & erythroid colony potentiating factor - promote

leucopoiesis & erythropoiesis &

- transforming growth factors (TGF), PDGF & fibroblast growth factors (FGF)

• Various enzymes like - collagenase, elastase, plasminogen activator etc -

participate in wound healing & tissue remodeling
• Monocytosis - TB, syphilis, leukemia

• Monocytopenia - hypoplastic BM
• Difference between phagocytosis by Neutrophil & Monocyte

Neutrophil Monocytes
- Antibacterial - Antibacterial & antiviral
- Engulf only smaller - Can engulf big organisms as well
bacteria
- Can engulf up to 20 - > 100 bacteria & viruses can be
bacteria engulfed
- They die after the - Do not die , remain as tissue
engulfment macrophages
• Macrophages –

- exact mechanism of
differentiation of monocyte to
macrophage is not known

- on becoming macrophage there

is increase in cell size, number
of cytoplasmic granules &
vacuoles
- average diameter - 25 to 50 µm, nucleus
eccentrically placed with one or two
nucleoli

- Cytoplasm contains multiple large

azurophilic granules
- contain all the surface receptors that are
present in monocytes

- In chronic tissue inflammations,

macrophages are converted into
multinucleated giant cells that are highly
phagocytic & microbicidal
• Dendritic Cells / Ag presenting
cells –

- Few monocytes are transformed

into highly specialized
mononuclear cells called
dendritic cells

- They are specialized in antigen

capture, rather than in
phagocytosis
- unlike macrophages, they lack
receptors for Ig, complements &
CSFs & specific granules in
cytoplasm

- hence they are weakly phagocytic

- present in blood & BM

- count 0.1 to 1 % of total

mononuclear cells
• Life span of WBCs -

• When active life span is less than when inactive

• Neutrophil - when inactive 24 hrs & when active less than 12 hrs, 7-10
days in tissues

• Eosinophil & Basophils - when inactive 8-12 days in tissues & when
active few hrs in blood
• Lymphocytes - from a few hours to a few years depending on whether it
is active or present inactive as a memory cell

• Monocytes – in circulation is 2-8 days & in tissues monocytes (now called

macrophage) can stay longer up to 20 days