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BLOOD and
       HEMOPOIESIS

Assoc. Prof. Dr. Karim Al-Jashamy
         IMS/MSU 2010
Formed Elements of Blood

• Erythrocytes (RBC)
• Leukocytes (WBC)
  – Granulocytes
  – Agranulocytes
• Platelets (Thrombocytes)
• Blood smear, stain using Wright.
• Aspirate bone (sternum, iliac crest) for bone
  marrow smear.
Functions of Blood
• Transport of:
    – Gases, nutrients, waste products
    – Processed molecules
    – Regulatory molecules
•   Regulation of pH and osmosis
•   Maintenance of body temperature
•   Protection against foreign substances
•   Clot formation
                                            19-3
Composition of
Blood
Plasma
• Liquid part of blood
  – Pale yellow made up of 91% water, 9% other
• Colloid: Liquid containing suspended
  substances that don’t settle out
  – Albumin: Important in regulation of water
    movement between tissues and blood
  – Globulins: Immune system or transport
    molecules
  – Fibrinogen: Responsible for formation of blood
    clots

                                                     19-5
Formed Elements
• Red blood cells (erythrocytes)
• White blood cells (leukocytes)
  – Granulocytes
     • Neutrophils
     • Eosinophils
     • Basophils
  – Agranulocytes
     • Lymphocytes
     • Monocytes
• Platelets (thrombocytes)

                                   19-6
Production of Formed Elements
• Hematopoiesis or hemopoiesis: Process of
  blood cell production
• Stem cells: All formed elements derived
  from single population
  – Proerythroblasts: Develop into red blood cells
  – Myeloblasts: Develop into basophils,
    neutrophils, eosinophils
  – Lymphoblasts: Develop into lymphocytes
  – Monoblasts: Develop into monocytes
  – Megakaryoblasts: Develop into platelets
                                                     19-7
Erythrocytes
         • Structure
            – Biconcave, anucleate
         • Components
            – Hemoglobin
            – Lipids, ATP, carbonic
              anhydrase
         • Function
            – Transport oxygen from
              lungs to tissues and
              carbon dioxide from
              tissues to lungs
                                 19-8
ERYTHROCYTE
• Numerous (5 x 106) /
  ml.
• Mature cell has no
  nucleus, organelles.
• Transports O2 and
  CO2.




SEM RBC
RBC

Biconcave shape increases surface area 20-30%.
Readily deform and pass through capillaries.
Cytoskeleton is unique. Spectrin is major protein.
Hemoglobin.


Energy from anaerobic respiration of glucose. (No
  mitochondria present)

Lifespan is 120 days. Removed by spleen and liver.

                                                     19-10
TEM RBC




Reticulocyte with a        RBC in capillary
few organelles.


                                              19-11
Hemoglobin
Hemoglobin (haemoglobin and abbreviated Hb or Hgb) is the iron-
    containing oxygen-tran also spelled sport metalloprotein in the
    red blood cells of vertebrates.

In mammals, the protein makes up about 97% of the red blood cell's
      dry content, and around 35% of the total content (including
      water).

Hemoglobin has an oxygen binding capacity of between 1.36 and
     1.37 ml O2 per gram of hemoglobin, which increases the total
     blood oxygen capacity seventyfold.



                                                                 19-12
 Hemoglobin is also found in outside red blood cells and
their progenitor lines.

 Other cells that contain hemoglobin include

   dopaminergic neurons in the substantia nigra,
   macrophages, alveolar cells, and mesangial cells in the
   kidney.


 In these tissues, hemoglobin has a non-oxygen carrying
function as an antioxidant and a regulator of iron
metabolism.                                           19-13
Hemoglobin




• Consists of:
  – 4 globin molecules: Transport carbon dioxide
    (carbonic anhydrase involved), nitric oxide
  – 4 heme molecules: Transport oxygen
      • Iron is required for oxygen transport
Normal values
Hemoglobin is measured in grams per deciliter of blood. The normal
levels are                           1.5-3 years - 12±.5
                                     5 y - 12.7±1
    Birth - 17±2                     10 y - 13.2±1
    1 day - 19±2                     Men - 15.5±1
    2-6 d - 19±2.5                   Women - 13.7
    14-23 d - 15.5±1                 Pregnant women: 11 to 12
    24-37 d - 14±2                   g/dl
    40-50 d - 13±2
                               Women: 12.1 to 15.1 g/dl
    2-2.5 month- 11.5±1
                               Men: 13.8 to 17.2 g/dl
    3-3.5 m - 11±1
                               Children: 11 to 16 g/dl
    5-7 m - 11.5±1
                               Pregnant women: 11 to 12 g/dl
    8-10 m - 11.7±.5
                                                                     19-15
    11-13.5 m - 12±.5
Erythropoiesis




• Production of red blood cells
   – Stem cells   proerythroblasts    early erythroblasts
     intermediate    late    reticulocytes
• Erythropoietin: Hormone to stimulate RBC production
                                                            19-16
Hematopoiesis




                19-17
Hemoglobin Breakdown




                       19-18
Leukocytes




             19-19
Leukocytes
• Protect body against
  microorganisms and
  remove dead cells and
  debris
• Movements
   –   Ameboid
   –   Diapedesis
   –   Chemotaxis
   –   Passive Immunity
   –   Active Immunity
   –   Antigen – Antibody

 Diapedesis: The movement or passage of blood cells, especially white blood cells,
 through intact capillary walls into surrounding body tissue
• Types
  – Neutrophils: Most common; phagocytic cells destroy
    bacteria (60%)

  – Eosinophils: Detoxify chemicals; reduce
    inflammation (4%)

  – Basophils: Alergic reactions; Release histamine,
    heparin increase inflam. response (1%)
  – Lymphocytes: Immunity 2 types; b & t Cell types.
    IgG-infection, IgM-microbes, IgA-Resp & GI, IgE-
    Alergy, IgD-immune response
  – Monocytes: Become macrophages                    19-21
NEUTROPHIL
Neutrophils, mature and almost!
Neutrophil Characteristics
• 60-70% of leukocytes
• diameter 10-12 µm
• nucleus 2-8 lobes

• chromatin in dense coarse lumps
• 'drumstick' on lobe in 3% of neutrophils in
  females (Barr body)
Neutrophils




Mature and younger
Cells (lobes)
                            Movement by pseudopodia.
                            Engulfed bacterium.
Neutrophils (Polymorphs)
• Produced in bone marrow. Granulocyte.
• Highly motile, phagocytic.
• Acute inflammatory response to tissue
  injury; ingest, destroy damaged tissue &
  bacteria.
• Lifetime activity consists of one burst of
  phagocytosis!
TEM Neutrophil
5 lobed nucleus.

Primary granules are lysosomes.
Secondary granules (secretory)
contain substances for
inflammatory processes (comple-
ment activation, leucocyte
adhesion,
bacterial cell wall lysis).
Few other organelles
(mitochondria).
Glycogen for glycolysis in O2-
depleted areas.
LYMPHOCYTE
Lymphocyte Characteristics
• 20-25% of leukocytes

• Diameter 6-8 µm.

• Nucleus spheroid or ovoid.

• Chromatin in dense lumps.

• Cytoplasm scarce and stained pale blue
Functions of Lymphocyte
• Central role in immunological defense.
• Most in circulating blood are inactive.
• Large lymphocytes (9-15m) are active B
  cells en route to tissues where they become
  plasma cells.
• T lymphocytes form in red marrow and
  move to thymus.
Lymphocytes




Large (activated) and small (inactive) lymphocytes.
Lymphocyte

A few mitochondria and
other organelles are present.
MONOCYTE
Monocyte Characteristics
• 3-8% of leukocytes.
• Largest leukocyte (20 µm).
• Nucleus indented and pale.
• Cytoplasm abundant and basophilic, non-
  uniform (foamy) appearance.
• Cytoplasm may contain a few fine
  azurophilic granules.
Functions of Monocytes
• Migrate to tissues and become microphages.
• Respond to necrotic material, invading
  microorganisms, and inflammation.
• Large content of hydrolytic enzymes.
• Great capacity for phagocytosis!
• Concept of a single functional unit, the monocyte-
  macrophage system consisting of Kupffer cells of
  liver, microglia of CNS, osteoclasts.
Monocytes


Mature cell has greater
Indentation of nucleus.
SEM Monocyte

Granules are similar to
lysosomes (acid phosphatase,
peroxidase).
Numerous golgi,
mitochondria, ribosomes.
BASOPHIL
Basophil Characteristics
• Less than 1% of leukocytes.
• Diameter 14 µm.
• Forms in red bone marrow.
• Nucleus large and bilobed.
• Chromatin is more finely textured so nucleus is
  more pale staining.
• Cytoplasm filled with large dark-blue staining
  granules (basophilic) which may obscure nucleus
  (blackberry appearance).
Basophil
Function: immunological response to
parasites. Contain many mediators of
inflammatory response. Closely related
 to mast cells. Basophils and mast cells
 bind to IgE produced in response to
allergens. Triggers rapid exocytosis of
granule contents (degranulation).

 This is the cause of immediate hypersensitivity reaction
characteristic of allergic rhinitis, some forms of asthma, urticaria,
anaphylactic shock).
Basophil




Bilobed nucleus
Granules (S) contain heparin, leukotrienes, histamine.
Mitochondria, ribosomes, glycogen in cytoplasm.
EOSINOPHIL
Eosinophil Characteristics
• Up to 5% of leukocytes.
• Diameter 12-15 µm.
• Nucleus usually bilobed.
• Chromatin clumped but not as dense as in
  neutrophil.
• Cytoplasm filled with numerous large
  eosinophilic (acidophilic) granules which
  stain pale-pink.
Functions of Eosinophils
• Phagocytic for antigen-antibody complexes.
• Defense against parasites.
• Release granules against parasites which are
  injured by enzymes.
• Undergo chemotaxis in response to bacteria
  but preferentially respond to basophils and
  mast cells.
SEM Eosinophil

Specific granules (S) stains
reddish.
S granules contain many
hydrolytic enzymes.
Contains glycogen, some
mitochondria, rER,
sER.
Thrombocytes

               • Cell fragments
                 pinched off from
                 megakaryocytes in red
                 bone marrow
               • Important in
                 preventing blood loss
                 – Platelet plugs
                 – Promoting formation
                   and contraction of clots


                                       19-46
Hemostasis

• Arrest of bleeding
• Events preventing excessive blood loss
  – Vascular spasm: Vasoconstriction of damaged
    blood vessels
  – Platelet plug formation
  – Coagulation or blood clotting



                                                  19-47
Platelets




Very complex with organelles but no nuclei.
Function: form plugs in damaged vessels, promote clot
formation, secrete substances that are involved in repair of
vessels.
Platelet Plug Formation




                          19-49
Coagulation
        • Stages
          – Activation of
            prothrombinase
          – Conversion of
            prothrombin to
            thrombin
          – Conversion of
            fibrinogen to fibrin
        • Pathways
          – Extrinsic
          – Intrinsic
                               19-50
Clot Formation




                 19-51
Fibrinolysis


          • Clot dissolved by
            activity of plasmin,
            an enzyme which
            hydrolyzes fibrin




                               19-52
Blood Grouping

• Determined by antigens (agglutinogens) on
  surface of RBCs
• Antibodies (agglutinins) can bind to RBC
  antigens, resulting in agglutination
  (clumping) or hemolysis (rupture) of RBCs
• Groups
  – ABO and Rh

                                              19-53
ABO Blood Groups




                   19-54
Agglutination Reaction




                         19-55
Rh Blood Group
• First studied in rhesus monkeys
• Types
  – Rh positive: Have these antigens present on
    surface of RBCs
  – Rh negative: Do not have these antigens present
• Hemolytic disease of the newborn (HDN)
  – Mother produces anti-Rh antibodies that cross
    placenta and cause agglutination and hemolysis
    of fetal RBCs
                                                      19-56
Erythroblastosis Fetalis




                           19-57
Diagnostic Blood Tests
            • Type and crossmatch
            • Complete blood count
               – Red blood count
               – Hemoglobin measurement
               – Hematocrit measurement
            • White blood count
            • Differential white blood
              count
            • Clotting

                                     19-58
Blood Disorders

• Erythrocytosis: RBC     • Hemophilia
  overabundance           • Thrombocytopenia
• Anemia: Deficiency of   • Leukemia
  hemoglobin              • Septicemia
  –   Iron-deficiency
                          • Malaria
  –   Pernicious
  –   Hemorrhagic         • Infectious
  –   Hemolytic             mononucleosis
  –   Sickle-cell         • Hepatitis

                                               19-59

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Anatomy

  • 1. BLOOD and HEMOPOIESIS Assoc. Prof. Dr. Karim Al-Jashamy IMS/MSU 2010
  • 2. Formed Elements of Blood • Erythrocytes (RBC) • Leukocytes (WBC) – Granulocytes – Agranulocytes • Platelets (Thrombocytes) • Blood smear, stain using Wright. • Aspirate bone (sternum, iliac crest) for bone marrow smear.
  • 3. Functions of Blood • Transport of: – Gases, nutrients, waste products – Processed molecules – Regulatory molecules • Regulation of pH and osmosis • Maintenance of body temperature • Protection against foreign substances • Clot formation 19-3
  • 5. Plasma • Liquid part of blood – Pale yellow made up of 91% water, 9% other • Colloid: Liquid containing suspended substances that don’t settle out – Albumin: Important in regulation of water movement between tissues and blood – Globulins: Immune system or transport molecules – Fibrinogen: Responsible for formation of blood clots 19-5
  • 6. Formed Elements • Red blood cells (erythrocytes) • White blood cells (leukocytes) – Granulocytes • Neutrophils • Eosinophils • Basophils – Agranulocytes • Lymphocytes • Monocytes • Platelets (thrombocytes) 19-6
  • 7. Production of Formed Elements • Hematopoiesis or hemopoiesis: Process of blood cell production • Stem cells: All formed elements derived from single population – Proerythroblasts: Develop into red blood cells – Myeloblasts: Develop into basophils, neutrophils, eosinophils – Lymphoblasts: Develop into lymphocytes – Monoblasts: Develop into monocytes – Megakaryoblasts: Develop into platelets 19-7
  • 8. Erythrocytes • Structure – Biconcave, anucleate • Components – Hemoglobin – Lipids, ATP, carbonic anhydrase • Function – Transport oxygen from lungs to tissues and carbon dioxide from tissues to lungs 19-8
  • 9. ERYTHROCYTE • Numerous (5 x 106) / ml. • Mature cell has no nucleus, organelles. • Transports O2 and CO2. SEM RBC
  • 10. RBC Biconcave shape increases surface area 20-30%. Readily deform and pass through capillaries. Cytoskeleton is unique. Spectrin is major protein. Hemoglobin. Energy from anaerobic respiration of glucose. (No mitochondria present) Lifespan is 120 days. Removed by spleen and liver. 19-10
  • 11. TEM RBC Reticulocyte with a RBC in capillary few organelles. 19-11
  • 12. Hemoglobin Hemoglobin (haemoglobin and abbreviated Hb or Hgb) is the iron- containing oxygen-tran also spelled sport metalloprotein in the red blood cells of vertebrates. In mammals, the protein makes up about 97% of the red blood cell's dry content, and around 35% of the total content (including water). Hemoglobin has an oxygen binding capacity of between 1.36 and 1.37 ml O2 per gram of hemoglobin, which increases the total blood oxygen capacity seventyfold. 19-12
  • 13.  Hemoglobin is also found in outside red blood cells and their progenitor lines.  Other cells that contain hemoglobin include dopaminergic neurons in the substantia nigra, macrophages, alveolar cells, and mesangial cells in the kidney.  In these tissues, hemoglobin has a non-oxygen carrying function as an antioxidant and a regulator of iron metabolism. 19-13
  • 14. Hemoglobin • Consists of: – 4 globin molecules: Transport carbon dioxide (carbonic anhydrase involved), nitric oxide – 4 heme molecules: Transport oxygen • Iron is required for oxygen transport
  • 15. Normal values Hemoglobin is measured in grams per deciliter of blood. The normal levels are 1.5-3 years - 12±.5 5 y - 12.7±1 Birth - 17±2 10 y - 13.2±1 1 day - 19±2 Men - 15.5±1 2-6 d - 19±2.5 Women - 13.7 14-23 d - 15.5±1 Pregnant women: 11 to 12 24-37 d - 14±2 g/dl 40-50 d - 13±2 Women: 12.1 to 15.1 g/dl 2-2.5 month- 11.5±1 Men: 13.8 to 17.2 g/dl 3-3.5 m - 11±1 Children: 11 to 16 g/dl 5-7 m - 11.5±1 Pregnant women: 11 to 12 g/dl 8-10 m - 11.7±.5 19-15 11-13.5 m - 12±.5
  • 16. Erythropoiesis • Production of red blood cells – Stem cells proerythroblasts early erythroblasts intermediate late reticulocytes • Erythropoietin: Hormone to stimulate RBC production 19-16
  • 17. Hematopoiesis 19-17
  • 19. Leukocytes 19-19
  • 20. Leukocytes • Protect body against microorganisms and remove dead cells and debris • Movements – Ameboid – Diapedesis – Chemotaxis – Passive Immunity – Active Immunity – Antigen – Antibody Diapedesis: The movement or passage of blood cells, especially white blood cells, through intact capillary walls into surrounding body tissue
  • 21. • Types – Neutrophils: Most common; phagocytic cells destroy bacteria (60%) – Eosinophils: Detoxify chemicals; reduce inflammation (4%) – Basophils: Alergic reactions; Release histamine, heparin increase inflam. response (1%) – Lymphocytes: Immunity 2 types; b & t Cell types. IgG-infection, IgM-microbes, IgA-Resp & GI, IgE- Alergy, IgD-immune response – Monocytes: Become macrophages 19-21
  • 24. Neutrophil Characteristics • 60-70% of leukocytes • diameter 10-12 µm • nucleus 2-8 lobes • chromatin in dense coarse lumps • 'drumstick' on lobe in 3% of neutrophils in females (Barr body)
  • 25. Neutrophils Mature and younger Cells (lobes) Movement by pseudopodia. Engulfed bacterium.
  • 26. Neutrophils (Polymorphs) • Produced in bone marrow. Granulocyte. • Highly motile, phagocytic. • Acute inflammatory response to tissue injury; ingest, destroy damaged tissue & bacteria. • Lifetime activity consists of one burst of phagocytosis!
  • 27. TEM Neutrophil 5 lobed nucleus. Primary granules are lysosomes. Secondary granules (secretory) contain substances for inflammatory processes (comple- ment activation, leucocyte adhesion, bacterial cell wall lysis). Few other organelles (mitochondria). Glycogen for glycolysis in O2- depleted areas.
  • 29. Lymphocyte Characteristics • 20-25% of leukocytes • Diameter 6-8 µm. • Nucleus spheroid or ovoid. • Chromatin in dense lumps. • Cytoplasm scarce and stained pale blue
  • 30. Functions of Lymphocyte • Central role in immunological defense. • Most in circulating blood are inactive. • Large lymphocytes (9-15m) are active B cells en route to tissues where they become plasma cells. • T lymphocytes form in red marrow and move to thymus.
  • 31. Lymphocytes Large (activated) and small (inactive) lymphocytes.
  • 32. Lymphocyte A few mitochondria and other organelles are present.
  • 34. Monocyte Characteristics • 3-8% of leukocytes. • Largest leukocyte (20 µm). • Nucleus indented and pale. • Cytoplasm abundant and basophilic, non- uniform (foamy) appearance. • Cytoplasm may contain a few fine azurophilic granules.
  • 35. Functions of Monocytes • Migrate to tissues and become microphages. • Respond to necrotic material, invading microorganisms, and inflammation. • Large content of hydrolytic enzymes. • Great capacity for phagocytosis! • Concept of a single functional unit, the monocyte- macrophage system consisting of Kupffer cells of liver, microglia of CNS, osteoclasts.
  • 36. Monocytes Mature cell has greater Indentation of nucleus.
  • 37. SEM Monocyte Granules are similar to lysosomes (acid phosphatase, peroxidase). Numerous golgi, mitochondria, ribosomes.
  • 39. Basophil Characteristics • Less than 1% of leukocytes. • Diameter 14 µm. • Forms in red bone marrow. • Nucleus large and bilobed. • Chromatin is more finely textured so nucleus is more pale staining. • Cytoplasm filled with large dark-blue staining granules (basophilic) which may obscure nucleus (blackberry appearance).
  • 40. Basophil Function: immunological response to parasites. Contain many mediators of inflammatory response. Closely related to mast cells. Basophils and mast cells bind to IgE produced in response to allergens. Triggers rapid exocytosis of granule contents (degranulation). This is the cause of immediate hypersensitivity reaction characteristic of allergic rhinitis, some forms of asthma, urticaria, anaphylactic shock).
  • 41. Basophil Bilobed nucleus Granules (S) contain heparin, leukotrienes, histamine. Mitochondria, ribosomes, glycogen in cytoplasm.
  • 43. Eosinophil Characteristics • Up to 5% of leukocytes. • Diameter 12-15 µm. • Nucleus usually bilobed. • Chromatin clumped but not as dense as in neutrophil. • Cytoplasm filled with numerous large eosinophilic (acidophilic) granules which stain pale-pink.
  • 44. Functions of Eosinophils • Phagocytic for antigen-antibody complexes. • Defense against parasites. • Release granules against parasites which are injured by enzymes. • Undergo chemotaxis in response to bacteria but preferentially respond to basophils and mast cells.
  • 45. SEM Eosinophil Specific granules (S) stains reddish. S granules contain many hydrolytic enzymes. Contains glycogen, some mitochondria, rER, sER.
  • 46. Thrombocytes • Cell fragments pinched off from megakaryocytes in red bone marrow • Important in preventing blood loss – Platelet plugs – Promoting formation and contraction of clots 19-46
  • 47. Hemostasis • Arrest of bleeding • Events preventing excessive blood loss – Vascular spasm: Vasoconstriction of damaged blood vessels – Platelet plug formation – Coagulation or blood clotting 19-47
  • 48. Platelets Very complex with organelles but no nuclei. Function: form plugs in damaged vessels, promote clot formation, secrete substances that are involved in repair of vessels.
  • 50. Coagulation • Stages – Activation of prothrombinase – Conversion of prothrombin to thrombin – Conversion of fibrinogen to fibrin • Pathways – Extrinsic – Intrinsic 19-50
  • 51. Clot Formation 19-51
  • 52. Fibrinolysis • Clot dissolved by activity of plasmin, an enzyme which hydrolyzes fibrin 19-52
  • 53. Blood Grouping • Determined by antigens (agglutinogens) on surface of RBCs • Antibodies (agglutinins) can bind to RBC antigens, resulting in agglutination (clumping) or hemolysis (rupture) of RBCs • Groups – ABO and Rh 19-53
  • 56. Rh Blood Group • First studied in rhesus monkeys • Types – Rh positive: Have these antigens present on surface of RBCs – Rh negative: Do not have these antigens present • Hemolytic disease of the newborn (HDN) – Mother produces anti-Rh antibodies that cross placenta and cause agglutination and hemolysis of fetal RBCs 19-56
  • 58. Diagnostic Blood Tests • Type and crossmatch • Complete blood count – Red blood count – Hemoglobin measurement – Hematocrit measurement • White blood count • Differential white blood count • Clotting 19-58
  • 59. Blood Disorders • Erythrocytosis: RBC • Hemophilia overabundance • Thrombocytopenia • Anemia: Deficiency of • Leukemia hemoglobin • Septicemia – Iron-deficiency • Malaria – Pernicious – Hemorrhagic • Infectious – Hemolytic mononucleosis – Sickle-cell • Hepatitis 19-59