Chapter 12 Leukopoiesis I.

Leukocytes y Defend organism against nonself agents y Direct and amplify phagocytic action through lymphokines Granulocytes y Granulocyte Pools Bone marrow, peripheral blood, endothelium of blood vessels, tissues y proliferation, maturation, storage y Proliferation myeloblasts, promyelocytes, myelocytes y capable of mitotic divisions y Maturation metamyelocytes and bands y no longer capable of mitosis but not yet fully functional y Storage bands and polymorphonuclear leukocytes y Marginating Pool adhere to vessel endothelium or egress to the tissue y Circulating Pool in the peripheral blood, counted in WBC count 1. Maturation: Stem Cell to Myeloblast y Earliest leukopoietic precursor Colony Forming Unit Spleen (CFU-S) or Hematopoietic Stem Cell (HSC) y CD (cluster designation) 34+ (signal transduction) thy1+ (cell cycling)

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II.

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HLA-DR (no class II HLA-DR antigens) Rh 123 (stain taken up by mitochondria) Dull (cell is not cycling) CFU-GEMM bone marrow- derived or myeloid cells CFU-GM (Colony Forming Unit GranulocyteMonocyte/Macrophage) Myeloblast earliest recognizable cells of neutrophilic series Interleukins and Colony Stimulating Factors (CSF) regulate cell numbers and function GM-CSF form granulocytes and monocytes/macrophages only G-CSF form granulocytes only M-CSF form monocytes/macrophages only

2. Neutrophil Maturation A. Myeloblast y From CD34 CD 13 and CD 33 y Less than 1% in bone marrow y 15-20 m y Nucleus is delicate with prominent nucleoli

Meager cytoplasm with RER, Golgi apparatus, and Azurophilic Granules y Granules color for myeloperoxidase y killing function is first to be operational y Incapable of motility, adhesion, and phagocytosis B. Promyelocyte (Progranulocyte) y 1% to 5% in bone marrow y 20 m (may be larger than myeloblast y May show slight clumping of nuclear chromatin pattern y Nucleoli begin to fade y Primary granules are dominant characteristic y Some motility may be present y Cathepsin G, elastase, proteinases killing C. Neutrophilic Myelocyte y Accumulation of secondary or Neutrophilic Granules y Last cell capable of mitosis y Longest stage of maturation y Less than 10% of marrow population y

Nuclear chromatin shows clumping y Nucleoli no longer visible y Dawn of neutrophilia (faint blush of pink) y Compounds within secondary granules thrombospondin receptor, -microglublin, apolactoferritin, lysozyme, plasminogen activators y Alkaline phosphatase and decay-accelerating factor y Motile D. Neutrophilic Metamyelocyte y Indented nucleus y Majors components to kill and degrade toxic, infectious, or nonself agents y Incapable of responding to chemotactic factors y 13% to 22% in bone marrow y Towards the end, Gelatinase Granule is made y Gelatinase granule composed of lysozyme and acetyl transferase E. Neutrophilic Band (Nonsegmented Form) y 40% in bone marrow y

Presence (segmented) or absence (nonsegmented) of nuclear segments y Band (C or S shaped), PMN (all other) y Band Nuclear Indentation (less than half of width), PMN Indentation (more than half of width) y Full motility, active adhesion properties, phagocytic ability y Band in peripheral blood less than 6% y Secretory vesicle store of surface membrane-bound receptors y Endocytosis F. Polymorphonuclear Neutrophil (Segmented Neutrophil) y Lobed nucleus y Active motility y Storage pool in bone marrow y 50% - 70% in peripheral blood y CD 15, CD 21, CD 3, CD 62, CD 11a, CD 18 y Stimulated by chemotactic factors y Egress of neutrophils into tissue diapedesis y

Phagocytosis/Pinocytosis 1. Recognition-Attachment Phase y Mediation by IgG y Actin, Myosin, binding proteins 2. Ingestion Phase (Phagocytosis) y Pseudopod extension y Microfilament rearrangement y Engulfment 3. Intracellular Kill: Hypertonic Mechanism y Potassium Influx 4. Digestion Phase: Degradation y Secondary lysosomal formation y Proteinases, hydrolases, arylsulfatases, phosphatises 5. Exocytosis y Removable of indigestible elements y Reversal of phagocytosis Eosinophil Maturation y y y Control mechanisms, IL3, IL5, GM-CSF, interferon CFU-GEMM CFU-Eo Myelocyte stage large, round granules containing crystalloid compound No alkaline phosphatase or lysozyme Large storage capacity in bone marrow Allergy or parasitic infections

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Nucleus that is in band form or bilobed Charcot-Leyden crystals result of eosinophilic degradation

Basophil Maturation y Irregularly-shaped granules y Deep purple to black y Granules heparin, chondroitin, sulfate, histamine, and other vasoactive and immunomodulatory factors y Membranes high affinity for IgE y Mast cell tissue equivalent of basophil Monocytes Monocyte/Macrophage Maturation A. Stem Cell to Monoblast y Primary site bone marrow y Secondary site spleen, reticuloendothelial sites y Strongly positive for CD33 y CD 4 T lymphocyte marker y Only function is mitosis y Eccentrically placed nucleus y 1 or 2 noticeable nucleoli B. Monoblast to Promonocyte y With some granulation y Folded, twisted, indented nuclei y Irregularly shaped cytoplasm

Motile and can participate in phagocytosis C. Promonocyte to Monocyte y Transport proteins, nonspecific inflammatory agents, storage materials, humoral-acting agents y Nucleus is slightly indented or curved y Chromatin is lacy with small clumps y Largest cell in peripheral blood y Cytoplasm with swirls of minute granules y Transitional cell D. Monocyte to Macrophage y Pleomorphic y Seen with pseudopods because of high motility y Function is phagocytosis E. Macrophage y Terminal cell of monocyte cell line y Free macrophages sites of inflammation and repair, alveolar spaces, peritoneal and synovial fluids y Fixed macrophages microglial cells, Kupffer cells, spleen, bone marrow, lymph nodes y Nucleus is round to reniform y 1 to 2 nucleoli y Lymphocytes

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Primary organs thymus and bone marrow Secondary sites Peyer s patches in GI tract, Waldeyer ring of tonsils, adenoids, lymph nodes Development y G proteins control diversity found in differentiation of T and B lymphocytes y Thymus T cells, primarily in bloodstream y Bone marrow B cells, primarily in lymph nodes y Plasma cell end cell of B lymphocyte maturation Lymphcyte Maturation A. Lymphoblast to Prolymphocyte y Round to oval nucleus y Loose chromatin, one or more active nucleoli y Scanty cytoplasm y Prolymphocyte more clumped chromatin, less prominent nucleoli, change in thickness of nucleolar membrane B. Prolymphocyte to Lymphocyte y Nucleus is round to oval

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Chromatin pattern is block type Rarest is large lymphocyte

Immunologic Differentiation y y y Tdt terminal deoxynucleotidyl transferase B cells T cells Tdt marker

Lymphocyte Activity: Regulation of Cellular Response y Regulation of immune function Morphology of Activation y Reactive lymphocyte less clumped nucleus, faintly stained multiple nucleoli seen, large in size, chromatin patterns less striking, elliptic to cleaved to folded nucleus, golgi apparatus seen, deeply basophilic cytoplasm