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WBCS classification, function,identification,structure


By: GautamSirvi (63 month(s) ago)

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WHITE BLOOD CELLS (WBCs) or LEUKOCYTES By ---Dr Raghuveer Choudhary Associate Professor Department of Physiology Dr S.N.Medical College,Jodhpur



Blood Cells:

3 Blood Cells RBCs, Red blood cells or erythrocytes WBCs, white blood cells or Leukocytes Platelets (thromobocytes)

Blood Components: Cells:

Blood Components: Cells Erythrocytes Red Blood Cells (RBC) O 2 & CO 2 transport White Blood Cells (WBC) Immune defense Phagocytosis Platelets: clotting

Blood Cells and Platelets:

Blood Cells and Platelets

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6 Cell Type Leukocytes ( lecuko - white) (White blood cells, WBCs) Description Spherical, nucleated cells Cells/mm 3 (µl) of blood 4000-11,000 Types Granulocytes Neutrophils Eosinophils Basophils Agranulocytes Lymohocytes Monocytes Leuckocytes


INTRODUCTION Leucocytes : Mobile units of the body ’ s defence mechanism Formed in the : Bone marrow Lymphoid tissue. Rapidly deployed through the blood to areas where: Infection & Inflammation are seen.

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Leukocytes (White Blood Cells) The leukocytes, also called white blood cells, They are formed partially in the bone marrow ( granulocytes and monocytes and a few lymphocytes ) and partially in the lymph tissue ( lymphocytes and plasma cells ).

Leukocytes (White Blood Cells) :

Leukocytes (White Blood Cells) After formation, they are transported in the blood to different parts of the body where they are needed in areas of serious infection and inflammation, thereby providing a rapid and potent defence against infectious agents.

Types of White Blood Cells. :

Types of White Blood Cells. Five types of white blood cells are normally present in the blood. They are 1. Polymorphonuclear neutrophils , 2. Polymorphonuclear eosinophils , 3. Polymorphonuclear basophils , 4. Monocytes , 5. Lymphocytes,


LEUCOCYTES: Types - Granulocytes - Neutrophil, 50-70% - Eosinophils, 1-4% - Basophils <1% - Agranulocytes - Monocytes 2-8% - Lymphocytes 20-40% - S & L Total leucocyte Count Adults= 4000-11000/c.mm of blood At Birth=20,000/c.mm


Leucopenia TLC <4000/ cmm Causes- Starvation,Typhoid fever,Viral infections,Bone marrow depression Irradiation of the body by x-rays or gamma rays , or exposure to drugs and chemicals that contain benzene or anthracene nuclei, is likely to cause aplasia of the bone marrow. Some common drugs, such as chloramphenicol (an antibiotic), thiouracil (used to treat thyrotoxicosis ), and even various barbiturate hypnotics, can cause leukopenia ,


Leucocytosis TLC >11000/ cmm Physiological cause Newborn Pregnancy,Lactation Exercise,Stress Pathological Causes Pyogenic Infections Burns Malignancy Allergic Reactions


Leukaemia A cancerous condition of blood in which TLC increases >50,000/ cmm & is associated with presence of immature WBCs in peripheral smear. which is usually characterized by greatly increased numbers of abnormal white blood cells in the circulating blood. T ypes of Leukemia . Leukemias are divided into two general types: lymphocytic leukemias and 2. myelogenous leukemias .

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Effects of Leukemia on the Body The first effect of leukemia is metastatic growth of leukemic cells in abnormal areas of the body. Leukemic cells from the bone marrow invade the surrounding bone , causing pain and a tendency for bones to fracture easily . Almost all leukemias eventually spread to the spleen, lymph nodes, liver, and other vascular regions , regardless of whether the origin of the leukemia is in the bone marrow or the lymph nodes. Common effects in leukemia are the development of infection, severe anemia , and a bleeding tendency caused by thrombocytopenia (lack of platelets) .


Granulocytes The first three types of cells, the polymorphonuclear cells, all have a granular appearance, for which reason they are called granulocytes, or, in clinical terminology, " polys ," because of the multiple nuclei.

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The granulocytes and monocytes protect the body against invading organisms mainly by ingesting them-that is, by phagocytosis . The lymphocytes and plasma cells function mainly in connection ith the immune system. .


N EUTROPHILS Most numerous Leucocytes ( 50 – 70%) Are 10 – 14 Microns in diameter. Have a constantly changing shape due to amoeboid movements. The Nucleus can have 1 – 6 lobes connected by a fine strand. The Cytoplasm contains fine granules.


N EUTROPHILS Nucleus - purple in colour , multi-lobed (2-6) connected by chromatin thread. Cytoplasm - Pinkish Granules –Fine, closely packed violet pink. Not seen seperately , do not cover nucleus.




Neutrophils Also called polymorphonuclear leukocytes 50–70% of circulating WBCs; 3,000 – 6,000/cumm. Purple cytoplasm granules with lysosomal enzymes and bactericides (hydrogen peroxide superoxide) Phagocytes that are the first to attack bacteria, engulf and digest pathogens with defensins Release prostaglandins and leukotrienes (inflammation and alarm call) Form pus 22

GRANULOCYTES : Neutrophils:

GRANULOCYTES : Neutrophils 50–70% of circulating WBCs; 3,000 – 6,000/ cumm . Size 10-14 micron Cytoplasm- purple in colour Granules – fine - Types - Pri /non-sp/ azuorophilic - Sec/sp/ n,e,b ( lectoferin,alkaline phosphatase ) - Contents – NADPH- Oxidase , SOD, Meyloperoxidase Nucleus – Color, Lobes, , Arneth Count, Shift I 5-10%, II 25-30%, III 45-47%, IV 16-18%, V 2% Functions - First line attack & kill bacteria by phagocytosis Pools – BM 90%, IV 3%, T 7%

Cytoplasmic Granules:

Cytoplasmic Granules -Fine, azurophilic (Stain with both Eosin & Methylene blue) in nature. Contain enzymes such as : Cathepsins . Phosphatases . Nucleases. Granules serve as lysosomes . MPO,


N EUTROPHILS & MONOCYTES: Functions They seek, attack and destroy invading bacteria, viruses and other injurious agents Neutrophils attack and destroy bacteria and viruses, even in the blood. Monocytes are immature until they enter the tissues. There, they swell up to 80 Microns, develop lysosomes , and become Macrophages, capable of defence .

Neutrophils & Macrophages:

Neutrophils & Macrophages Diapedesis : They squeeze through the pores of the blood vessels. Amoeboid movement : They move at rates several times their own length! Chemotaxis : Directed movement – cells move to wards infected areas.


N EUTROPHILIA Increased neutrophil count >7400/ cmm , can be due to: Physiological- Exercise, Pregnancy,Lactation , Menstruation Pathological- Acute Pyogenic Infections,Tissue destruction,M.I ., Trauma,Surgery , Burn,DKA,Lithium


Neutropenia Decrease in neutrophils <2500/ cmm Physiological- In infancy (40%) Typhoid fever, Viral infections, BM depression, Malaria, Kala- azar,Measels , Hepatitis, Anti-thyroid drugs, Anti- inflamatory drugs, Anti convulsant drugs Chloramphenicol,cotrimoxazole .


EOSINOPHILS 1– 4% of the Leucocytes. Size-10-14 micron Have a typical ‘ Spectacle shaped ’ , bilobed nucleus. Have coarse bright pink staining granules in the cytoplasm.


EOSINOPHIL Nucleus - Blue-violet,2-3 lobed,often bilobed Cytoplasm - Eosinophilic,light pink- red,granular Granules -Large coarse,uniform size, brick red to orange, seen seperately , do not cover nucleus


Eosinophils >400/ cmm Eosinophilia or increased count occurs in: P arasitic infestations.( Hook worm,Ascariasis ) A llergic conditions.( Bronchial asthma,Urticaria,Hay fever) T PE: Tropical Pulmonary Eosinophilia . S kin disorders- Eczema,contact dermatitis,pempigus , EOSINOPHILS





Eosinophils :

Eosinophils Also called acidophils 1–4% of circulating WBCs;150-300/cumm. Acidophilic cytoplasm, pink in colour Attack large parasites by excreting toxic compounds Sensitive to allergens Control inflammation with enzymes that counteract inflammatory effects of neutrophils and mast cells 35

GRANULOCYTES : Eosinophils:

GRANULOCYTES : Eosinophils Size & number 10-14 micron, 150-300/ cmm Cytoplasm- Acidophilic cytoplasm, pink in colour Granules - coarse bright pink staining granules in the cytoplasm. - Contents - MBP, - ECP, , Histaminase & Arylsulfatase Nucleus Spectacle shaped ’ , bilobed nucleus. Functions - Mild Phagocytic - Attack & kill Parasites by enz ,( MBP, Eo2, ECP, ) - Reduce ill effects of allergic inflammation by detoxifying infl. Subs. released by mast cells - phagocytosing Ag- Ab complexes.

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Eosinophils are weak phagocytes , and they exhibit chemotaxis , but in comparison with the neutrophils , it is doubtful that the eosinophils are significant in protecting against the usual types of infection. Eosinophils , however, are often produced in large numbers in people with parasitic infections, and they migrate in large numbers into tissues diseased by parasites.

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Eosinophils attach themselves to the juvenile forms of the parasite and kill many of them. They do so in several ways: (1) by releasing hydrolytic enzymes from their granules, which are modified lysosomes ; (2) probably by also releasing highly reactive forms of oxygen that are especially lethal to parasites; and (3) by releasing from the granules a highly larvacidal polypeptide called major basic protein .


Basophils Are very few in number: < 1%. Have a large indented nucleus which is obscured by cytoplasmic granules. Granules are coarse and basic staining: blue . They are abundant and protrude through the cell membrane.


GRANULOCYTES : Basophils Features – Size 10-14 micron ,10-100/ cmm Nucleus -Blue violet,irregular shape,not clearely seen because overlaid by granules. Cytoplasm — basophilic,bluish,granular Granules –Large Very Coarse , variable size, deep purple coloure , seen seperately , completely fill the cell & cover the nucleus Contents - His, Brk , Sero , Hep , Lysoenz , ECF-A


GRANULOCYTES : Basophils Functions - Release Natural anticoagulant Heparin [activate LPL] - Role similar to Mast cells - Role in some allergic reactions mediated by IgE (Immediate Hypersensitivity Reactions) & - Releasing His, Brk , Sero , Hep , Lys Enz to produce local vas. & tissue allergic reactions. - Release ECF-A which phagocytose ag-ab complex & dampens allergic reactions

Basophils :

Basophils Less than 1% of circulating WBCs;10-100/cumm. Slightly basophilic cytoplasm, appears blue Plenty of granules are present which obscure the nucleus boundary. Small cells that accumulate in damaged tissue Release histamine to dilate blood vessel and heparin prevent blood clotting Similar to mast cells (found in the tissues) 43




Basophilia Count >100/ cmm Small pox Chicken pox Polycythemia vera Hypersensitivity reactions Haemolytic reactions CML


AGRANULOCYTES : Monocytes Features – 2–8% of circulating WBCs, 300-600/ cumm Size -10-18 microns(1.5-3 times of RBC) - Nucleus- Blue violet,large single,May be indented,hourse shoe shape,or kidney shape, eccentric in position Cytoplasm - Abundent,more or equal to nucleus amount, light blue [C/N Ratio]50:50 Granules - absent

Monocytes :

Monocytes 2–8% of circulating WBCs, 300-600/cumm Largest WBC, Light blue staining, single, kidney shaped nucleus which is eccentric in position Enter peripheral tissues and become macrophages Engulf large particles and pathogens Secrete substances that attract immune system cells and fibroblasts to injured area 48




Function- Monocytes Active phagocytosis Enter in tissues to become tissue macrophases Kill tumour cells Form imp. IL-1,TNF2,Compliments,transferin ,proteases, acid hydrolases Monocytosis count >800/ cmm TB,Typhoid,Monocytic leukemia,Malaria


LYMPHOCYTES: Morphologically, LARGE Lymphocytes: Sized about 12 – 15 µ Thin cytoplasmic rim Large spherical nucleus No cytoplasmic granules.


LYMPHOCYTES: Small Lymphocytes: Sized about 7-9 µ. ( Smallest Leucocytes) Thin cytoplasmic rim & Large spherical nucleus. No granules visible . Gives ink spot appearance

Large and small lymphocyte in a smear:

Large and small lymphocyte in a smear 53


LYMPHOCYTES Features - Size & Number 1500-2700/ cmm - - Cell Types - Small & Large, B & T Cell [NK, Helper, Suppressor & Memory Cells] - Functions - Immune Reactions, Hypersensitivity Reactions & Immuno-regulation



Lymphocytes :

Lymphocytes T cells, B cells and NK cells 20–40% of circulating WBCs;1500-2700/cumm Note the small amount of cytoplasm Small, just slightly larger than RBCs Migrate in and out of blood Most of them are in connective tissues and lymphatic organs (spleen, lymph nodes) Respond to specific antigens 57



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Produced in bone marrow Lymphocytes Remain & mature in B one Migrate & mature in T hymus Lymphocyte T cell B cell

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Mature B & T cells migrate to lymphoid organ

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Physiological Classification : T and B ‘ T ’ LYMPHOCYTES : Thymus trained or schooled cells Responsible for Cell mediated immunity . Provide protection against intracellular pathogens LYMPHOCYTES :


Trained in the Bone marrow(Bursa Fabricius in birds) Responsible for Humoral Immunity.(Immunity through Antibody production) Protect the body from encapsulated pyogenic bacteria like Pneumococcus & streptococci. LYMPHOCYTES :

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Thymus Spleen Tonsil Lymph Node Bone Marrow Lymphoid Organs 1 2 3 4 5

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Lymphoid Organ


LYMPHOCYTES: The only human WBCs whose site of development is not just BM, but also tissues referred to as primary and secondary lymphoid organs In humans, the primary lymphoid organs are the thymus and bone marrow, the secondary organs include the spleen, Peyer ’ s patches of the GI tract, the Waldermyer ring of the tonsils and adenoids, the lymph nodes and modules scattered throughout the body


Lymphocytosis In children (60%) Chronic infections Lymphatic Leukemia Diptheria Mumps Hepatitis Viral infections


WHITE BLOOD CELLS(LEUKOCYTES) Leukocytes : have nuclei and other organelles, not involved in oxygen transport. Functions: Defend against pathogens Remove toxins and wastes Attack abnormal cells WBC in blood vs. tissue Very small numbers in blood : 4000 to 11000 per microliter Outnumbered 700:1 by RBCs 68

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Only 1% of WBC are in blood Most WBCs are not found in blood but instead in connective tissue proper and in lymphatic system organs. Circulating WBCs WBCs can migrate out of bloodstream into tissues Have amoeboid movement (using actin) Attracted to chemical stimuli (positive chemotaxis) Some are phagocytic: neutrophils, eosinophils and monocytes. 69

Life Span of the White Blood Cells :

Life Span of the White Blood Cells The life of the granulocytes after being released from the bone marrow is normally 4 to 8 hours circulating in the blood and another 4 to 5 days in tissues where they are needed .

Life Span of the White Blood Cells :

Life Span of the White Blood Cells The monocytes : Have a short transit time, 10 to 20 hours in the blood, before wandering through the capillary membranes into the tissues. Once in the tissues, monocytes swell to larger sizes to become tissue macrophages , and, in this form, can live for months unless destroyed while performing phagocytic functions. These tissue macrophages are the basis of the tissue macrophage system, which provides continuing defense against infection.

Life Span of the White Blood Cells :

Life Span of the White Blood Cells Lymphocytes : Enter the circulatory system continually, along with drainage of lymph from the lymph nodes and other lymphoid tissue. After a few hours, they pass out of the blood back into the tissues by diapedesis . Then, still later, they re-enter the lymph and return to the blood again and again; thus, there is continual circulation of lymphocytes through the body. The lymphocytes have life spans of weeks or months; this life span depends on the body's need for these cells.

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How White Blood Cells Enter the Tissue Spaces: By Diapedesis . Neutrophils and monocytes can squeeze through the pores of the blood capillaries by diapedesis . 2. by Ameboid Motion. Both neutrophils and macrophages can move through the tissues by ameboid motion. 3. Attracted to Inflamed Tissue by Chemotaxis . Many chemical substances in the tissues cause both neutrophils and macrophages to move toward the source of the chemical. This phenomenon is known as chemotaxis .

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How White Blood Cells Enter the Tissue Spaces

Phagocytosis :

Phagocytosis The most important function of the neutrophils and macrophages is phagocytosis , which means cellular ingestion of the offending agent. Phagocytes must be selective of the material that is phagocytized ; otherwise, normal cells and structures of the body might be ingested.

Phagocytosis :

Phagocytosis B. Whether phagocytosis will occur depends especially on three selective procedures .: First most natural structures in the tissues have smooth surfaces , which resist phagocytosis . But if the surface is rough, the likelihood of phagocytosis is increased. Second, most natural substances of the body have protective protein coats that repel the phagocytes. Conversely, most dead tissues and foreign particles have no protective coats, which makes them subject to phagocytosis . Third, the immune system of the body develops antibodies against infectious agents such as bacteria. The antibodies then adhere to the bacterial membranes and thereby make the bacteria especially susceptible to phagocytosis .

Phagocytosis :

Phagocytosis C. The Complement 3 molecules attach to receptors on the phagocyte membrane, thus initiating phagocytosis . This selection and phagocytosis process is called opsonizatio n .

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Phagocytosis is the ingestion of microorganisms or other matter by a cell. Many white blood cells engulf invasive microorganisms by the process of phagocytosis . The steps in phagocytosis are: 1 . Chemotaxis is the process by which phagocytes are attracted to microorganisms. 2 . Attachment : The phagocyte then adheres to the microbial cell. This adherence may be facilitated by opsonization – coating the microbe with plasma proteins. 3 . Ingestion : Pseudopods of phagocytes engulf the microorganism and enclose it in a phagosome to complete ingestion. 4. Digestion : Lysosomes fuse with the phagosome to form a digestive vacuole. The microbe is killed and digested.

Role of Phagocytosis:

Role of Phagocytosis The engulfment, digestion, and subsequent processing of microorganisms by macrophages and neutrophils . 1- Chemotaxis & attachment a- Attraction by chemotact . Subst. ( microbes , inflam . tissues) b- Attachment by receptors on surfaces of phagocytes 2- Ingestion * Phagocytic pseudopodia surround organism forming phagosome * Opsinins and co factors enhance phagocytosis * Fusion with phag . granules and release digestive ,toxic contents

Role of Phagocytosis:

Role of Phagocytosis 3- Killing (two microbicidal routes) a- Oxygen depended system (powerful microbicidal agents) Oxygen converted to superoxide, anion, hydrogen peroxide , activated oxygen and hydroxyl radicals .( Myeloperoxidase ) b- Oxygen-independent system (anaerobic conditions) Digestion and killing by lysozyme. Lactoferrin , low pH, cationic proteins and hydrolytic and proteolytic enzymes

Second Line of Defense:

Second Line of Defense Figure 16.8a Phagocytosis

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.A single neutrophil can usually phagocytize 3 to 20 bacteria before the neutrophil dies. Phagocytosis by Macrophages. Macrophages are the end-stage product of monocytes that enter the tissues from the blood. They are much more powerful phagocytes than neutrophils , often capable of phagocytizing as many as 100 bacteria. They have the ability to engulf much larger particles ,

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Also, after digesting particles, macrophages can extrude the residual products and often survive and function for many more months . O nce Phagocytized , Most Particles Are Digested by Intracellular ENZYMES. Both neutrophils and macrophages contain an abundance of lysosomes filled with proteolytic enzymes especially geared for digesting bacteria and other foreign protein matter. 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.

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