Lecture 4 - dont study Flashcards
what is the reference range of leukocytes and how many types do we study from the PBS
3.6-10.6x10^9/L
we look at 5 basic types in wrights stain
how are leukocytes identified
-through antigen typing - they have surface antigens s
-surface antigens are confirmed with antibodies are tagged by a detection system reaction with cell membrane proteins
-by immunophenotyping of blood cells by membrane antigen
-detected using fluorescent dyes or enzymes
-based on CD# or cluster of differentiation
-CD34= stem cell can make CD4 = helper t lymph, CD 8 = cyto toxic tlymph, and cd3 - activated t lymph
-flow cyto - in hem
-immunohistochemistry
where do leukocytes develop from
-hemaotopoietic stem cells in the bone marrow
- after differentiation and maturation they are released into peripheral circulation and then into tissues via kinetics
-the number of circulating cell lines varies based on gender, age, ethnicity, and state of leukocytes (stresses)
-their main function is mediating immunity (ADAPTIVE OR INNATE)
how do leukocytes act in innate immunity
NON SPECIFIC
-cells that defend against other orgs
-recognize and respond in generic manner
-does NOT provide long lasting immunity
PHAGOCYTOSIS BY NEUTROPHILS, MONOCYTES AND MACROPHAGES
how do leukocytes act in adaptive immunity
SPECIFIC
-specialized cells that eliminate pathogens or prevent their growth
-creates immunological memory to specific pathogen
-enhances response to subsequent exposure
PRODUCTION OF ANTIBODIES TO SPECIFIC ANTIGENS BY LYMPHOCYTES AND PLASMA CELLS
neutrophils and monocytes have common progenitor
GMP
G-CSF
M CSF
-neutrophils and monocytes have common progenitor - GRANULOCYTE- MONOCYTE PROGENITOR -GMP
G-CSF- granulocyte -colony stimulating factor - major cytokine responsible for stimulating NEUTROPHIL production
M-CSF -major cytokine responsible for stimulating Monocyte production
Eosinophils and Basophils share
a common progenitor
Eosinophils and Basophils share
a common progenitor called EOSINOPHIL- BASOPHIL PROGENITOR EBP
Interleukin-3, -5 (IL-3 and IL5) and GM-CSF- cytokines responsible for stimulating eosinophil production
Interleukin-3 (IL-3) and Thymic stromal lymphoprotein (TSLP)– cytokine responsible for stimulation of basophil production
what happens as a granulocyte matures
- the N:C ration decreases
-nuclear chromatin gets coarser a and the nucleoli disappears
-the nucleus starts to change shape
- cytoplasm starts to get less blue
-presence and type of granules - none, primary or 2ndary
how do Promyelocyte look?
-high n:c ratio 3:1
-large round oval nucleus
-fine chromatin - small clumps
-more cytoplasm than a blast
-blue/basophilic cytoplasm
-ABUNDANCE OF PRIMARY GRANULES RED OR AZUROPHILIC (non specific with anti bacterial properties)
-1-3 nucleolus
-divides
-HAS PRIMARY OR NON SPECIFIC GRANULES
BIGGER ONE
how do myeloblasts look ?
-high n:c ratio 4:1
-large round oval nucleus
-multiple nucleoli
-fine chromatin - no clumps
-little cytoplasm
-blue/basophilic cytoplasm
-absent or 20 primary granules
-2-5 nucleolus
-divides
BIGGER ONE
what does a Myelocyte look like
-n:c ratio depends on cell line
-round to OVAL nucleus- can be flat or a weird
-nucleoli not visible -FAINT
-more clumped than pro
-cytoplasm less basophilic -pink lavender
-loss of primary granules
-SECONDARY OR SPECIFIC GRANULES APPEAR IN THE CYTOPLASM (eos, baso, neut)
-gradual transition from pro - myelocyte (there are early and late myelocytes)
-LAST DIVIDING STAGE
-the number of 2ndary granules increases as the cell matures
-if you say myelocyte ALONE then it refers to the neutrophilic cell line
-has a FOCAL PERINULEAR CLEARING
what appears in a neutrophilic myelocyte
- DAWN OF NEUTROPHILIA
-appearance of secondary granules in Golgi apparatus area
-2ndary granules give the neut cytoplasm a LAVENDER pink colour
-these granules increase in number and spread throughout the cytoplasm as the cell matures
what appears in a metamyelocyte
n:c ratio depends on the cell line
-indented nucleus in the shape of a kidney bean or peanut
-nucleoli are not visible
-moderately clumped
-cytoplasm only has 2ndary granules with very little basophilia
-FOCAL PERINULEAR CLEARING IS COMMON
NO MORE DIVISION
what do band cells look like
- n:c depends on the line
-nuclear indentation is greater than 1/2 the width of the nucleus (pinched but not segmented)
-nucleus is thin sausage shaped S or C - BUBBLE LETTERS
-chromatin coarse clumped
-abundant 2ndary granules
What are the stages (pool) of Neu maturation
-neuts most numerous in Bone Marrow and Peripheral blood
-Neuts in BM released into the peripheral blood
1) - Stem cell pool - self renewing, pluripotent stem cells - Hematopoietic stem cell
2)Proliferation (MITOTIC) pool - consists of cells that are dividing CMP-GMP-MYELOBLAST- PROMYELOBLAST- MYELOCYTE
3) Maturation (storage) pool - cells that are undergoing maturation make up the BM reserve are ready for release - METAMYELOCYTE TO MATURE NEUTROPHIL
-stages earlier than Band neut are not normally found in the PB earlier form may be seen in severe leukemia (only metamye or myelo)
-infection severity in indicated by increase in band forms - report under LEFT SHIFT
PATH from stem cell pool to myeloblast
stem cell pool - cmp with IL3 - to GMP with GM-CSF - to myeloblast with IL3, GM-CSF, G-CSF
what is the function on neuts in the PB
–innate immunity
-phagocytosis and destruction of foreign material and microorganisms
-half CIRCULATE free in CIRCULATING POOL while the other half adhere to the vessel wall in Marginal (capillary walls of liver, spleen , and lung)
-in PB the half life is 7 hours then they migrate into tissues where their half life can be a few hours or longer if they are fighting infection /inflammation
what is the function of neutrophil granules
-lysosomes containing many proteins and enzymes
-lysosomes can merge with phagosomes to kill engulfed bacteria
Primary granules in Pro & early Myelocytes
‒ Myeloperoxidase
‒ Defensins
‒Elastase
secondary, tert and scretory granules from late myelo or segmented nuet stage have
‒ Lactoferrin
‒ Alkaline phosphatase
‒ Lysozyme
‒ Gelatinase
what is the function of eosinphils
-no phagocytosis
–half life of 18 hours
-secrete cytokines to stimulate other cells like antigen presenting cells to promote proliferation of T cells
- initiate type 1 or type 2 immune response
-can be increased in parasitic infections - kills Helminths via secretion of 2ndary granules
-increased allergic reactions (hallmark finding)
-best studied in asthma
-eos can invade and damage through degranulation the respiratory and GI tract mucosa
what is the function of eosinophil granules
-eos can form 1mary , 2ndry granules, small lysosomal granules, lipid bodies and storage vesicles
-release granule content into extracellular spaces (degranulation -MBP CATIONIC PROTEIN AND GROWTH FACTOR NGF
-2ndry granules have various enzymes and presynthesized proteins like cytokines or chemokines
-MAJOR BASIC PROTEIN, ACID PHOSPHATASE, LYSOZYME, CATALASE, GM-CSF
what is the basophil function
-poorly understood function - it will enter the tissues via PB with half life of 60 hours
–2ndary granule contents not fully understood but production is changed based on activation signals
-has histamine , IL3, IL4, vascular growth factors , HEPARIN SULFATE
-can release large amounts of cytokines
-regulate T2H cells and indue B cells to produce IGE
-initiates allergic inflammation and in hypersensitivity reactions
Mast cells are similar - tissue effector cells of allergic and inflammatory reactions
they enter a mucosal surface from BM before maturing
monocytes in BM, PB and PB TO TISSUE
- in BM there are few monocytes
- no storage pool and are released in PB right after maturing
-reservoir of immature mono in spleen
in PB - there is a pool along vessel walls - small circulating pool
PB to Tissue - migrate to lots of tissue sites and turn into MARCOPHAGES
liver, spleen, lungs , skin, intestines BONE MARROW
what are the resident macrophages in
Liver
lungs
brain
skin
spleen
intestines
peritoneum
bone
synovial
kidneys
reproductive
lymph nodes
Liver - KUPFFER CELLS
lungs - ALVEOLAR MACROS
brain - MICROGLIA
skin -LANGERHANS CELLS
spleen -SPLENIC MACRO
intestines -INTESTINAL MACRO
peritoneum -PERITON MACRO
bone -OSTEOCLASTS
synovial-TYPE A CELL
kidneys - RENAL MACRO
reproductive - REPROD ORG MAC
lymph nodes - DENDRITIC CELLS
what does a macrophage look like
-bigger than monocyte
-oval eccentric nucleus
-reticulated chromatin
-vacuolated cytoplasm - with debris of phagocytized cells or orgs
Monocyte/Macrophage Function in innate immunity, adaptive and housekeeping
INNATE Stimulate phagocytosis and release cytotoxins (for
example, nitric oxide) against:
‒ Bacteria, Viruses
‒ Fungi, Parasites
‒ Tumor cells
Adaptive Act as an antigen-presenting cell by exposing Activate B-Lymphs and T-Lymphs to initiate adaptive immune response
Housekeeping
Destruction of senescent RBCs
● Removal of debris or dead cells
● Maintenance of iron storage pool for erythropoiesis
● Synthesis of proteins (such as, coagulation factors, complement components, interleukins, growth factors, and enzymes
what are they 3 group divisions of lymphopoiesis
- B cells (adaptive immunity)
- T cells (adaptive immunity) - MOST
- Natural Killer (NK) cells (innate immunity)
‒ Only a small % of Lymphs are NK
● Lymphocytes make up 20- 40% circulating WBCs
what are the two FUNCTIONAL CATEGORIES that lymphs can be put into
- Produce antibodies (Humoral Immunity):
‒ B cell Lymphocytes - Attack foreign organisms or cells (Cellular immunity):
‒ T cell Lymphocytes
‒ NK cells
how does B cell maturation take place
-lymphoid progenitors develop in the BM (they are not exposed to antigen but have IG antigen receptors)
-these cells then travel into 2ndry lymph orgs like germinal centerss of lymph nodes) where the cells are then exposed to antigens - look like resting lymphs
-after being exposed to antigens they transform into plasma (EFFECTOR ) cells and memory cells
-essential for antibody production and presenting antigens to T cells
what is a plasma cells
- final stage of B cell lymph differentiation
-produces IMMUNOGLOBULIN OR ANTIBODIES against antigens
-NOT SEEN IN PB unless in disease state - on PBS oval cell, round nuc, perinuclear hof (halo), dark blue cytoplasm
-in PBS you can see B cells that have transformed with dark blue cytoplasm called PLASMACYTOID LYMPHOCYTE
how does T cell maturation take place
- lymphoid progenitors migrate from BM to thymus
- cell MATURE in thymus by making t cell receptors or surface markers unique to each T cell
- t cells then MIGRATE to 2NDARY LYMPH ORGS or pb where they are exposed to antigens causing activation, transformation into memory cells or effector T cells
what to T lymphs look like
● CD surface antigens develop as T cells mature into specific types in thymus
● Main ‘markers’ are: CD4 and CD8
● CD4+ T cells are often called Helper T cells
● CD8+ T-cells are called Cytotoxic T cells
what are reactive lymphs
- Cytotoxic T - Lymphocytes (CD8+)
-Activation and transformation after antigen exposure seen in disease conditions or viral infections like mononucleosis
-the cell is very large, lots of spreading, dark blue cyto, big nuc,
-cell is then capable of division
what are Natural Killer Lymphocyte
Maturation:
● either the BM or Thymus
● Non-specific immunity – (innate immunity) capable of killing tumor cells or virally infected
cells without prior exposure or sensitization
● Modulate function of Macrophages and T cells
Morphology:
● Large cell with increased cytoplasm
● Cytoplasm contains azurophilic (burgundy) granules
● Similar morph to Cytotoxic T cells
Monopoiesis
Stages include:
1. Monoblast
2. Promonocyte
3. Monocyte
4. Macrophage
