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ARNETH COUNT_1

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ARNETH COUNT_1

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WHITE BLOOD CORPUSCLES

 Mobile units of body’s protective system


 Soldiers defending against microorganism
 Seek out & destroy foreign invaders
 Phagocytosis
 Forming Ab & sensitised lymphocytes
 Differ from RBC
 Formed in bone marrow & lymphoid organs
 Average lifespan is short
 Myeloid : Erythroid – 75% : 25%
CLASSIFICATION

WBC

Granulocytes Agranulocytes

Neutrophil Eosinophil Basophil Monocyte Lymphocyte


NORMAL VALUES
Total count – adults – 4000 – 11,000 cells/cu.mm.
- at birth 10,000 – 25,000 cells/cu.mm.
Cell Differential Absolute count
Count (cells/cu.mm.)
Neutrophil 50 – 70% 2000 – 7000
Eosinophil 1 – 4% 50 – 500

Basophil 0 – 1% 10 – 100

Monocyte 2 – 8% 200 – 800

Lymphocyte 20 – 40% 1500 - 4000


Genesis of WBC
Pluripotent Stem Cell

Myelocytic Lineage Lymphocytic Lineage

Myeloblast Monoblast Lymphoblast

Promyelocyte Promonocyte Prolymphocyte

Myelocyte Monocyte Lymphocyte

Metamyelocyte

Band forms

Mature Granulocyte
LEUCOPOIESIS
Myeloblast
12-18µm;large ,pale nucleus; no granules;
2-5 nucleoli +;mitosis +;no motility
Promyelocyte
Size ↓;round nucleus; primary granules+;
No nucleolus
Myelocyte
Nucleus small & flattened; coarse chromatin;
cytoplasm extensive; primary & secondary granules+
Metamyelocyte
Nucleus indented; mitosis -;more of sec granules
than primary granules; amoeboid movement +
Band metamyelocyte

Mature granulocyte
SUMMARY OF GRANULOPOIESIS

Decrease in cell size


Differentiation
Appearance of specific granules
Appearance of nuclear lobes
Loss of divisibility
Development of functional ability
FACTORS AFFECTING
GRANULOCYTE MACROPHAGE –
COLONY STIMULATING FACTOR (CSF-
GM)
CSF-G, CSF-M
INTERLEUKINE-1,3,6 (IL1, IL3, IL6)
TUMOR NECROSIS FACTOR (TNF)
NEUTROPHIL
 Size-10 – 14 µm.
 DC – 50 – 70%
 In children, N 20%<adults
 Microphage actively
phagocytic.
 7 hrs – blood 4-5 days-tissue
 Multilobed nucleus
 Fine purple granules in pink
cytoplasm.
Secondary granules
Primary granules Lactoferrin – Fe binding Protein
Lysozyme Cationic Bactericidal Protein
Peroxidase VIT. B12 Binding protein
ARNETH COUNT
* Neutrophil count based on lobes of nucleus
* Indicates stages of development
* N1-5%,N2-30%,N3-45%,N4-18%,N5&6-2%
* N1 +N2+N3 >80% - Shift to left
* Regenerative shift
Hyperactive bonemarrow (pyogenic infection,
haemorrhage)
* N4 +N5+N6>20% - Shift to right
* Degenerative shift
Hypoactive bone marrow (pernicious anemia, aplasia
of bone marrow)
ARNETH COUNT

N1 N2 N3

N4 N5
EOSINOPHIL
• Size 10 -14 µm
• DC - 1-4%
• Bilobed nucleus
• Life span 8-12 days
• Eosin coloured coarse granules in cytoplasm
• Granules obscure cytoplasm
(Major Basic Protein , Eosinophilic Cationic Protein,
Histaminase, Arylsulphatase, Eosinophil peroxidase,
Lysophospholipase)
• Functions
Defends body from allergic reactions
(Neutralise allergen, larvicidal, parasiticidal)
Weakly phagocytic
Limits the effects of antigen antibody reactions
BASOPHIL
 Size 10 – 14 µm
 DC- 0-1%
 Similar to mast cells
 Multilobed, large, irregular nucleus
 Deep blue coarse cytoplasmic granules
 Granules obscure nucleus & cytoplasm
(Heparin, Histamine, Slow reacting substance)

Functions:
 Phagocytic
 Heparin-prevents intravascular clotting
 Histamine & SRS-take part in IgE mediated allergic
reactions along with mast cells.
MONOCYTES
♦ Size 10-18µm
♦ DC - 2-8%
♦ Large, pale cells
♦ Pale staining round / indented
eccentric nucleus
♦ Pale blue & clear cytoplasm, fine
azur granules
♦ Remain in circulation -10-20 hrs
♦ Go to tissues – swell up - Tissue
macrophages - survive for 3
months – continual defence –end
as multinucleated giant cells
FUNCTIONS OF MONOCYTES

 Actively phagocytic – Mononuclear macrophage-


engulf 100 bacteria
 Participate in immune response –process and
present antigen to T & B lymphocyte
 Monocytes on stimulation secrete IL -1,
promotes proliferation and maturation of T
lymphocytes
 Destroy senescent RBC - initiate catabolism of
Hb
LYMPHOCYTES
Small (80%) 7-10 µm
Large (20%)10-14 µm
• 20-40 %
• Round non granular cells
• Large round centrally placed nucleus
• Formed in bone marrow & in lymph organs
T- Thymus (80%)
B- Bursal equivalents(20%)
Site of Formation --- Circulation---
Tissues
Life span - weeks to months
FUNCTIONS –
• Mediate immune responses
T- Cellular Immunity
B- Humoral Immunity
Applied aspects
Neutrophilia

Physiological Pathological

Exercise
Acute pyogenic infections
Pregnancy
Rheumatic fever
Parturition Myocardial infarction
Emotional stress
↓ Neutropenia

Physiological Pathological

Typhoid
Rare Paratyphoid
Exposure to cold Aplastic anemia
Viral infection -measles
Monocyte

Monocytosis ↓ Monocytopenia

Malaria
Rare
Kala-azar
Bone marrow failure
Monocytic leukemia
Aplastic anemia
Basophil

Basophilia ↓ Basophilopenia

Chronic myeloid leukemia Rare


Polycythemia Aplastic anemia
Lymphocyte

Lymphocytosis ↓ Lymphocytopenia

Chronic infections Immunosuppressive therapy


Infectious Mononucleosis ACTH therapy
Lymphoma Hodgkin’s disease
Viral infection Bone marrow failure
DEFENSIVE FUNCTIONS OF
NEUTROPHIL AND MONOCYTE
NEUTROPHIL
Attack and destroy bacteria ,virus & injurious agents , Can
destroy even in circulation
MONOCYTES
A little ability to kill in circulation
Become macrophages (80 µm) in tissue
- Actively phagocytic
PHAGOCYTOSIS – Cellular ingestion of offending agent

Basis of selection
Rough surface
No protective protein coat
Antibodies & Complement adhere to bacterial membrane
STEPS INVOLVED IN
DEFENCE
 Margination
 Diapedesis
 Chemotaxis
 Opsonisation
 Phagocytosis
• Phagosome formation
• Degranulation
• Respiratory Burst
• Discharge of myeloperoxidase
• Digestion by intracellular enzyme
 Walling off effect
MARGINATION
Neutrophils attracted to endothelial surface by
SELECTINS
Bind to neutrophil adhesion molecules of INTEGRIN
family

DIAPEDESIS
Squeeze through pores < cell size
involves microtubules and microfilaments
AMOEBOID motion- Velocity 40µm per min

OPSONISATION
Complements and antibodies attach to bacteria and
render them tasty
Facilitates phagocytosis
CHEMOTAXIS
 Chemoattractants
Bacterial toxins
Degenerative products of inflamed tissues
Reaction products of activated complement
Reaction products caused by plasma clotting
 Chemotactic substances attract Neutrophil &
Monocyte to infected area
 Depend on concentration gradient
 Greater near source
CHEMOTAXIS & PHAGOCYTOSIS
DIAPEDESIS
Phagocytosis
Capture, Ingest & Destroy
• Neutrophils – Microphages
• Monocytes - Macrophages
• Projects pseudopodia
• Enclose bacteria
• Invaginates
• Breaks from cell membrane
• Fusion of lysosome with phagosome
• Exocytosis of granules
• Digestion by intracellular enzymes
PHAGOCYTOSIS
PHAGOCYTOSIS
Inflammation
o Vasodilation
o Capillary permeability
o Clotting of fluid in the interstitium
o Migration of Granulocytes & Monocytes
o Swelling of tissue cells
Defence mechanisms
 Tissue macrophages (within minutes)
 Neutrophils (within hours)
 Monocytes from blood (within hours –
days)
 Granulocytes & Monocytes (days –
weeks)
Differences
Neutrophils Macrophages
Can phagocytose 5 -20 Even 100
bacteria
Begins phagocytosis A little ability to kill in
immediately circulation
Less powerful More powerful
Particles >bacteria Can engulf even RBC
Shorter lifespan Can extrude residual
Die in the process products & can live for
more months
Activated by immune
system
Do not die
RETICULOENDOTHELIAL SYSTEM
Monocyte macrophage system(MMS)
Mononuclear phagocytic system (MPS)
 Generalised phagocytic system
 Special group of cells scattered in different parts
• Powerful phagocytes
• Stained by supravital stains
 Monocytes – macrophages
 Liver – Kupffer cells
 CNS – Microglia
 Lungs – Alveolar macrophages
 Skin – Langerhans cells
 Bone – Osteoclasts
 Lymph nodes, spleen & bone marrow – Dendritic cells
 Connective tissue – Histiocytes [Fixed & wandering]
Functions of RES
 More powerful phagocytic action than Neutrophil
 Engulf larger particle
 Engulf more numbers (100)
 Survive even after ingestion of bacteria
 Ingest & destroy RBC & release bilirubin
 Also destroy dead WBC’s & platelets
 Ingest & process antigen & present it to Lymphocyte(play a role in
Immunity)
 Produce IL-1-activates T cell
CSF-GM-regulates erythropoiesis
 Initiate development of antibody
 Activated by lymphokines of T lymphocytes.Migrate to area of
inflammation & engulf bacteria
 Sites of hemopoiesis during fetal life(spleen & liver)
Functions of Spleen
 Haemopoietic organ
 Culling – Destroying RBC’s B4 lifespan
 Pitting – Removing Heinz bodies
 Platelet destruction
 Reservoir of blood
 Splenectomy – prone for malaria
APPLIED ASPECTS
Reticulosis
lymphadenopathy Infectious mononucleosis
splenomegaly Hodgkin’s lymphoma
hepatomegaly

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