Module 6 WBC Maturation and Function Acknowledgments Ministry of Health Guyana Centers for Disease Control and Prevention CDC Global AIDS Program GAP Guyana Centers for Disease Control and Prevention CDC Atlanta American Society for Clinical Pathology ASCP Objectives Review the origin of blood cells using the present stem theory of Haematopoiesis Describe factors involved in the stimulation and regulation of leukopoiesis granulocytes monocytes and lymphocytes including Cytokines colony stimulating factors interleukins produced locally Cells which produce cytokines Differentiate the maturation sequence of granulo cytes monocytes and lymphocytes based on Cell size Nuclear size and nuclear chromatin Cytoplasmic color and presence absence of granules Objectives Describe the lifespan appearance and function of the following leukocytes Granulocytes neutrophils eosinophils basophils Monocytes macrophages Lymphocytes including plasma cells List the contents of primary and secondary granules in granulocytes Discuss the granulocyte pools including changes caused by physiologic stimuli List the contents of the granules in monocytes Objectives Describe the sequence of phagocytosis Differentiate antigen independent and antigen dependent lymphopoiesis including the location of primary and secondary lymphoid tissues Discuss monocyte and lymphocyte roles in the immunologic response Discuss neutrophil and monocyte roles in the inflammatory response Objectives Correlate the lymphoid cell type with the following cluster designation CD markers CD2 3 T cells including CD4 and CD8 cells CD19 20 B cells Correlate the following reactive changes in neutrophil and lymphocyte morphology with associated conditions Toxic neutrophils toxic granulation Dohle bodies vacuoles Reactive lymphocytes Stem Cell Theory of Haematopoiesis Pluripotential Haematopoietic stem cells give rise to two multipotential progenitor cells The myeloid stem cell ultimately produces unipotential progenitor cells committed to a particular cell line with receptors for specific growth factors CFU E Erythropoietin regulates red cell production CFU Meg Thrombopoietin regulates platelet production CFU GM bipotential GM CSF colony stimulating factor primarily regulates granulocyte and monocyte production The lymphoid stem cell gives rise to cells committed to B or T cell lineages Lymphocyte production is influenced by environment bone marrow or thymus and several interleukins Stem Cell Theory of Haematopoiesis Normal Bone Marrow Full of developing precursor cells in all stages of maturation Bone Marrow Megakaryocyte 1000x Leukopoiesis Rate of leukocyte production is adjusted to need and requires an intact bone marrow Precursor cells divide mature to end stage cells Controlled by cytokines produced by T lymphocytes and monocytes macrophages Cytokines may be lineage specific or act on multiple cell lines GM CSF IL 3 Only mature cells are released normally Leukocytes perform defense functions in tissues Granulocyte lifespan is 1 2 days Monocytes and lymphocytes are long and short lived Classification of Leukocytes Type of defense Phagocytes inflammatory response foreign not self Immunocytes immune response Site of origin Myeloid or lymphoid stem cell Shape of nucleus Presence or absence of granules Immunophenotype using monoclonal antibodies CD19 20 Lymphoid B cells CD2 3 Lymphoid T cells CD4 helper T cells or CD8 suppressor T cells CD Surface Antigens Granulocyte Maturation Includes neutrophils eosinophils basophils Bone Marrow Four stages granules appear Primary non specific granules are lysosomes coated by lipid that contain lytic enzymes peroxidase lysozyme Secondary specific granules neutrophil eos baso Marrow Blood Immature band form National Guard Most numerous bone marrow precursors short lifespan Regulated by GM CSF Normal to see a low of band neutrophils in the blood Blood Mature segmented form Granulocyte Maturation Basophil secondary granules Primary non specific granules Neutrophil secondary granules 1 2 3 4 5 6 Eosinophil secondary granules Abbott Manual Granulocyte Maturation Eosinophils basophils are not staged low Immature stages myelo meta band are neutrophils Primary granules appear in promyelocyte stage Primary granules are present in mature cells but don t stain after myelocyte stage under normal conditions Seg N Myeloblast No granules Band Neutrophil Primary granules present don t stain Promyelocyte Primary granules present stain Blood Abnormal Seg N Granulocyte Maturation Specific secondary granules B Basophil N Neutrophil E Eosinophil Kidney shaped Myelocyte B Myeloblast Promyelocyte Myelocyte N Myelocyte E E Band B Metamyelocyte N Band N Band E Segmented B Segmented N Segmented Granulocyte Maturation Myeloblast Promyelocyte Seg N N Myelocyte Blood Abnormal N Metamyelocyte Promyelocyte Promyelocyte Blood Abnormal Band N Granulocyte Maturation Band N Eos Baso Seg N Seg N Blood Eos Baso Baso Band N Band N Granulocyte Maturation Broken Neutrophil Segmented neutrophil Broken Eosinophil Hypersegmented Neutrophil 5 lobes Eosinophil Megaloblastic anemia pernicious anemia Granulocyte Pools IMPORTANT SLIDE National Guard Soldiers Granulocyte Function Neutrophils Major role is phagocytosis IgG C3 receptors Defend against bacterial infections arrive 1st die Granules contain lytic enzymes alkaline phosphatase Seg neutrophils are 1 WBC type in normal adults Increased band neutrophils in the blood left shift Necrotic Neutrophil Pyknotic nucleus Band Neutrophils Granulocyte Function Seg neutrophil Ingested Bacteria Erythrophagocytosis Ingested RBCs Eosinophils Control parasitic infections Modify allergic reactions caused by basophils Basophils Immediate hypersensitivity rxns anaphylaxis Granules contain histamine heparin Granulocyte Function Baso Band N Seg N Eos Baso Seg N Mast cell Bone marrow Exploded Baso Monocyte Maturation Regulated by GM CSF Bone Marrow Two stages Marrow Blood Monocyte Largest normal cell in the blood Enter tissues progressively enlarge to become fixed or free macrophages of the RE system Spleen liver bone marrow lymph nodes lungs Abbott Manual 1 2 3 Monocyte Maturation Monoblast Blood Promonocyte Monocyte Tissue Macrophage Bone Marrow Monocyte Tissue Monocyte Macrophage Monocyte Function Role in defense Phagocytic cells receptors for IgG C3 lytic enzymes Arrive after neutrophil do not die Contain lipases to control TB infections Monocyte RES functions Splenic macrophages Bone marrow macrophages Kupffer cells