Leukocytes D.Rezazadeh Department of Medical Laboratory Science Kermanshah,Faculty of Paramedical.
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Transcript of Leukocytes D.Rezazadeh Department of Medical Laboratory Science Kermanshah,Faculty of Paramedical.
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Leukocytes
D.RezazadehDepartment of Medical Laboratory Science
Kermanshah,Faculty of Paramedical
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RBC Membrane Proteins
peripheral alpha & beta
spectrin bands 1 & 2
actin integral (lots of
sialic acid) glycophorin A band 3
inorganic anion transport channel
cation pump regulates Na+/K+ at ratios 1:12/25:1
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RBC Membrane Lipids
phospholipids lipid bilayer
unesterified cholesterol
facilitates selective cation permeability
1:1 equilibrium w/plasma cholesterol
increases in membrane cholesterol = target cells!!
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4
HEMATOPOIETIC FACTORS
NAME CELLULAR SOURCE CELL TYPES PRODUCED IN
INCREASED NUMBERS
Erythropoietin (EPO) Kidney cells, Kupffer cells
rbc
G-CSF Monocytes, fibroblasts, endothelial cells
n
M-CSF Monocytes, fibroblasts, endothelial cells
m
GM-GSF T cells, monocytes, fibroblasts, endothelial
cells
n, m, e, meg, rbc
IL-1 Macrophages, endothelial cells, fibroblasts
n, m, e, b, meg, rbc
IL-3 T cells n, m, e, b, meg, rbc
IL-4 T cells b
IL-5 T cells e
IL-6 Macrophages, endothelial cells, fibroblasts
n, m, e, b, meg, rbc
n, neutrophils; m, monocytes; e, eosinophils; b, basophils; meg, megakaryocytes; rbc, red blood cellsIL – interleukin , CSF – colony stimulating factor
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NEUTROPHILS
Myelopoiesis - the production and development of myeloid cells in the bone marrow Growth factors (colony-stimulating
factors - CSF’s) and interleukins regulate blood cell development by:
Mediating proliferation Differentiation into cell lines Maturation of hematopoietic progenitor
cells
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NEUTROPHILS
Three series of granulocytes (named for colors taken up by granules in Wright’s stain) Neutrophils
Granules stain equally with eosin and methylene blue
Normal in circulation = 42 - 75% Eosinophils
Granules stain mainly red Normal in circulation = 0-4%
Basophils Granules stain mainly with methylene blue Normal in circulation = 0-1%
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Maturation of the neutrophilic series
Myeloblast Size: 10-20µm Cytoplasm
Slight amount/blue
No granules Nucleus
Round/Central or eccentric
Reddish purple
1-3 nucleoli Fine meshlike
chromatin with no clumping
N:C ratio = 4:1
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Maturation of the neutrophilic series
Promyelocyte (progranulocyte)
Size: 10-20µm Cytoplasm
Increased amount/blue Fine, azurophilic,
nonspecific granules present
Nucleus Round/Central or
eccentric Reddish-purple Fine, meshlike
chromatin beginning to clump
1-2 nucleoli N:C ratio = 3:1
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Maturation of the neutrophilic series
Myelocyte (last myeloid cell capable of division)
Size: 10-18µm Cytoplasm
Increased, bluish-pink (“dawning of neutrophilia”)
Fine azurophilic, specific granules
Nucleus Round or
oval/eccentric Reddish-purple Chromatin loosely
clumped No nucleolus
N:C ratio = 2:1 or 1:1
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Maturation of the neutrophilic series
Metamyelocyte (also called “meta” or “juvenile”)
Size: 10-18µm Cytoplasm
Moderate amount Specific granules
(neutrophilic, eosinophilic or basophilic)
Nucleus Indented (kidney-bean
shaped) Central or eccentric Purple Clumped chromatin No nucleolus
N:C ratio = 1:1
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Maturation of the neutrophilic series
Band Size: 10-16µm Cytoplasm
Moderate amount Specific granules
(see meta) Nucleus
Elongated, narrow band (sausage) shape with uniform thickness
Central or eccentric Clumped, coarsely
granular chromatin
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WBC'S (LEUKOCYTES)
NEUTROPHILS: Most abundant 50-70%
Function: phagocytosis granules with hydrolytic
enzymes defensins - antibiotic
proteins,
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EM: specific granule: 70% small, 0.3-0.4 um ovoid or irregular in shape contain: lysozyme, defensin azurophilic granule: 30% large, round or ovoid electron dense lysosome: acid phosphatase, peroxidase,
acidic hydrolase
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---function: phagocytose bacterium: specific g.-kill B azurophilic g.- digest B---life span: in blood 1 day, in Tissue 2-3
days
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Granule type Protein
specific granules (or "secondary granules")
alkaline phosphatase, lysozyme, NADPH oxidase, collagenase, Lactoferrin and Cathelicidin
azurophilic granules (or "primary granules")
myeloperoxidase, bactericidal/permeability-increasing protein (BPI), Defensins, and the serine proteases neutrophil elastase and cathepsin G
tertiary granules cathepsin and gelatinase
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WBC'S (LEUKOCYTES) EOSINOPHILS: 2-4%
of WBC's Function: Allergies,
parasitic worms granules w/lysosomes
and digestive enzymes ( enzymes for bacteria)
attack parasitic worms - release enzymes onto the worms
lessen allergies by phagocytizing immune (Ab/antigen) complexes
inactivate certain inflammatory chemical
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EM: granules: round or ovoid with cube-liked electron dense crystal contain: -ACPase-histaminase-peroxidase
---function: counteract the infection of parasite reduce allergic reaction---life span: in blood 18 hour, in Tissue 6-12
days
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WBC'S (LEUKOCYTES)BASOPHILS: 0.5% of WBC'S Function: liberate heparin,
histamines histamine containing
granules = inflammatory chemical that vasodilates
attracts other WBC's to site of inflammation
mast cells - like basophils are found in Tissue = tissue basophils?
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WBC'S (LEUKOCYTES) LYMPHOCYTES: 25-
40% of WBC'S Function: immune
responses and memory, mainly found in lymph tissue
T cells - immune response to viral infected and tumor cells
B cells - give rise to plasma cells which produce Ab's
more later
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EM: /free ribosome /mito. /RER---classification: TC: thymus dependent lymphocyte, 75%,
involve in cellular immune reaction and regulate immune response
BC: bone marrow dependent lymphocyte, 10-15%, become into plasma cell, involve in humoral immune response
large granular cell: KC (killer cell) and NKC( natural killer cell), 10%
---function: involve in immune response
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WBC'S (LEUKOCYTES)
MONOCYTES: 3-8% of WBC's Function: phagocytosis in tissues differentiate into macrophages evident in chronic infections -
Tuberculosis defense vs. viruses and certain bacteria activate lymphocytes
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EM: azurophilic granule: lysosome- contain
peroxidase, ACPase,non-spacial esterase and lysozyme
---function: actively mobile and chemotaxis---life span: in blood 1-5 days, in Tissue
which become into macrophage- mononuclear phagocytic system, MPS
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3) blood platelet: cytoplasmic fragment of megakaryocyte in bone marrow
---normal number: 150-450X109/L ---structure: LM: disc-liked → irregular in
shape Stimulation
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EM: /cell coat: glycosaminoglycan and glycoprotein/tubular systems: opening tubule system- increasing the exchange
area, facilitate the intaking and releasing dense tubule system- SER, collecting Ca2+ and
synthesizing prostaglandin/granules: specific granule:, with dense core, contains
clotting factor, fibrinogen, acidic hydrolase dense granule: contains serotonin(5-
hydroxytryptamine), ADP,ATP, Ca2+ and adrenalin
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---function: involve in clotting and stopping the
bleed: -aggregation -release components of granule,
make protrombin become into thrombin, then later make fibrinogen become into fibrin
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* stromal cells: ---including reticular cell, fibroblast,
macrophage, endothelial cell and lipid-laden cell
---function: supporting secrete cellular factors, regulate the
proliferation and differentiating of hemopoietic cells
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Maturation of the neutrophilic series
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Function of the neutrophilic series
Neutrophils - defense against bacterial infections
Eosinophils Regulate inflammation Respond to antigenic stimulation in chronic
allergies Interact with larval stages of some helminthic
parasites Basophils
Histamine in granules plays a role in acute, systemic allergic reactions (sudden release of histamine can cause anaphylactic shock)
Granules also contain heparin
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MONOCYTES
Monopoiesis - the production and development of monocytes, macrophages and their precursors. Monocytes are produced in the bone marrow. Their primary function is phagocytosis.
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MONOCYTES
Other names for macrophages Histiocytes in the loose connective tissue Kupffer cells in the liver Osteoclasts in the bone Microglial cells in the nervous system Also scattered throughout the body in all organ
tissues (spleen, lung, abdomen, etc.) One monocytes have grown into macrophages, they do not normally reenter the bloodstream except in cases of inflammation
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Maturation of the monocytic series
Monoblast Size: 12-20µm Cytoplasm
Blue, moderate amount
No granules Nucleus
Round, oval or slightly folded
Eccentric Reddish-purple Chromatin fine and
thready Vacuoles may be
present 1-2 nucleoli
N:C ratio = 4:1
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Maturation of the monocytic series
Promonocyte (difficult to discern) Size: 21-20µm Cytoplasm
Paler gray-blue and abundant “Bleb-like” pseudopods at periphery May contain fine red “dust-like” particles Vacuoles may be present
Nucleus Round with chromatin creases or brain-like folding Central Reddish-purple with light clumping of chromatin 0-2 nucleoli
N:C ratio = 3:1 or 2:1
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Promonocyte
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Maturation of the monocytic series
Mature monocyte (Normal in circulation: 2-9%) Size: 15-18µm Cytoplasm
Pale gray-blue and abundant Bleb-like pseudopods at periphery, variable shape Numerous fine pale red dustlike granule scattered
throughout Vacuoles common
Nucleus Increased folding or elongated Central Purplish with finer clumped chromatin No nucleoli
N:C ratio = 2:1 or 1:1
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Monocyte
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LYMPHOCYTES
Normal adult value: 20-45% Produced in the bone marrow
(primary lymphoid tissue)
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LYMPHOCYTES
Growth factors cause lymphs to differentiate. They proliferate and mature in the primary lymphoid organs:
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LYMPHOCYTES
Thymus - T cells Function
Cellular immune response (they do not produce antibodies) Regulate antibody reactions by helping or suppressing
activation of B lymphs. T cells act as “helper cells” (HIV or AIDS virus causes a destruction of these helper cells), which are instrumental in aiding B cells in antibody production, and “suppressor cells”, which act as a “thermostat” to shut off the system or keep it under control. The helper to suppressor ratio is very important and is normally 2:1.
Protect against viral, bacterial, fungal or protozoan infection Responsible for chronic organ graft rejection. (T cells have
many unique antigens on their cell surfaces, some of which are the HLA - human lymphocyte antigen - markers. Cytotoxic and helper T cells act against these HLA markers if they are foreign to the host organism. HLA markers are instrumental in chronic graft rejection and that is why tissue typing is done to establish a good donor match.)
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LYMPHOCYTES
Bone marrow - B cells Function
Humoral (antibody) immune response by transforming into antibody-producing plasma cells
Defense against encapsulated bacteria such as streptococci
Mediate hyperacute organ rejection
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Secondary lymphoid tissue Lymph node, spleen, Peyer’s patches in
intestine and mucosal tissues such as tonsils
Active immune response occurs in above tissues where lymphs communicate and interact with antigen-presenting cells, phagocytes and macrophages
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Maturation sequence of lymphocytes
Lymphoblast (which is preceded by a lymphoid stem cell)
Cell size: 10-20µm Cytoplasm
Blue/scanty No granules, Auer
rods are never present
Nucleus Purple, smooth
chromatin Round, central or
eccentric 1-2 nucleoli
N:C ratio = 4:1
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Maturation sequence of lymphocytes
Prolymphocyte (difficult to distinguish)
Size: 9-18µm Cytoplasm
Blue, scanty Usually granules are
absent, but a few azurophilic granules may be present
Nucleus Purple, condensed
chromatin Round or indented,
eccentric 0-1 nucleoli
N:C ratio = 4:1
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Maturation sequence of lymphocytes
Mature lymphocyte Cell size: 7-10µm (a typical
normal lymph has a nucleus that is the size of a normal RBC)
Cytoplasm Light blue, scanty to
moderate Few azurophilic granules
may be present Nucleus
Purple, dense, clumped chromatin
Round or indented, eccentric
No nucleoli
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Large lymphocytes versus monocytes
Size Large lymph: 12-15µm Mono: 15-18µm
Nucleus Large lymph: clumped, condensed Mono: lacy, brainlike folds
Granules Large lymph: large azurophilic, easy to count Mono: red, fine
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Maturation sequence of lymphocytes
Variant lymphs Other terms used are reactive, atypical, Downey
cell, transformed, virocyte, plasmacytoid Caused by antigenic stimulants such as viruses,
post-transfusion reactions and organ transplants
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Reactive lymphocyte
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Plasma cells
Plasma cells Function is the synthesis and excretion of antibodies
(immunoglobulins) Normally not present in the peripheral blood; comprise
2% of bone marrow cells. (May be seen in the peripheral blood in the disease called multiple myeloma, a disease of uncontrolled production of immunoglobulins.)
End stage of the B lymphocyte Appearance
Size: 10-18µm Cytoplasm is dark blue with perinuclear halo and may contain vacuoles
indicating antibody synthesis Nucleus is round, eccentric, dark purple with dense clumped chromatin
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Plasma cells
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Thank you
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Sites of Haemopoiesis Yolk sac
Liver and spleen
Bone marrow Gradual replacement
of active (red) marrow by tissue inactive (fatty)
Expansion can occur during increased need for cell production
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Extravascular Erythrocyte Destruction Normal end-of-life
event Inherited RBC Defects
Membrane abnormalities
Enzyme deficiencies Hemoglobinopathies Thalassemias
Acquired RBC Defects B12 Deficiency
Immune-mediated destruction
Drug-induced Autoimmune
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IntravascularErythrocyte Destruction Intravascular hemolysis
Normal end-of-life event
Complement Activation PNH PCH
Physical/mechanical Trauma
DIC Prosthetic heart
valves Toxins
Arsenic poisoning Bacterial Infections
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Lab Evidence of Hemolysis
Extravascular Urine urobilinogen Fecal urobilin Yellow-colored
plasma
Intravascular Increased Reticulocyte
Index Decreased haptoglobin Decreased hemopexin Presence of
Methemoglobin/methalbumin
Increased indirect bilirubin Increased urobilinogen Decreased RBC Increased iron stores
Hemosiderinuria Ferritin
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C/EBPα,ε G.CSF نوتروفیلC/EBPα,ε,β IL5 ائوزینوفیل
IL3,Kit ligand بازوفیلPU.1 , C/EBPβ M.CSF مونوسیت
PAX5,IKAROS IL4 لنفوسیتBGATA_3,notch1 IL7,IL2 لنفوسیتT
C/EBPγ SCF,IL15 NKسلGATA_1,NFE2 EPO,GMCSF,IL3 اریتروئید
GATA_1,GATA_2 TPO,IL3,IL11,IL6 مگاکاریوسیتیک
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Lymphocytes and Lymphoid Tissues
Primary lymphoid tissues
(thymus and bone marrow), Secondary lymphoid tissues
(lymph nodes, Peyer’s patches, and spleen) Tertiary lymphoid tissues
(skin and mucosal)
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Thymus
Cortex
Medulla
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Thymicprocessingof T-cells
See notes
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