GRANULOPOIESIS

NEUTROPHIL DEVELOPMENT

 Neutrophils are present in the peripheral blood in two forms according to whether the
nucleus is SEGMENTED or still in BAND shape.
 Segment Neutrophils make up the vast majority of circulating leukocytes.
MYELOBLAST
Size: 15-20 um in diameter  Occupies about four-
fifths of the cell.
Cytoplasm
 NC ratio of 4:1
 Small amount in relation
to the rest of the cell  Extremely fine chromatin
pattern
 Usually moderate blue in
color  Reddish purple in color
 Texture is smooth and  Contains two to five
usually nongranular nucleoli

Nucleus
 Round or slightly oval
PROMYELOCYTE
Size: 15-21 um in diameter Nucleus
Cytoplasm  Occupies half or more of
the cell
 Pale blue to basophilic
(full of primary azurophilic  NC ratio of 3:1 to 2:1
granules)
 Oval or round
 Contains a few to many,
 Coarser chromatin
large blue to reddish
pattern
purple staining
nonspecific (primary)  2-3 nucleoli present
granules
MYELOCYTE
Size: 12-18 um in diameter  Oval or round
Cytoplasm  Chromatin pattern
becomes coarser and
 Moderate amount
more condensed
 May contain few patches
 No nucleoli present
of blue
 Nucleus may be
 Few to moderate
eccentric
number of nonspecific
granules  NC ratio 1:1
 Small, specific
(secondary) granules
appear
Nucleus
MYELOCYTE

The specific granules appear first as

dirty orange to blue. These
Pink specific granules may be granules are larger than the specific
seen as pinkish or lighter staining and nonspecific granules of the
areas in the cytoplasm neutrophil.
METAMYELOCYTE
Size: 10-15 um in diameter  Indented or kidney
shaped
Cytoplasm
 Chromatin pattern is
 Moderate to abundant
coarse and clumped and
amount giving a
stains dark purple
decreased NC ratio
 Few nonspecific granules
 Full complement of
specific granules
 Synthesis of tertiary
granules (gelatinase
granules)
Nucleus
BAND
Size: 9-15 um in diameter Nucleus
Cytoplasm  Elongated or band-shape
 Moderate to abundant  Deeply indented from
amount giving a the metamyelocyte stage
decreased NC ratio
 Chromatin pattern is
 Few nonspecific granules coarse and clumped
 Full complement of
specific granules
SEGMENTED NEUTROPHIL
Size: 9-15 um in diameter
Cytoplasm Nucleus
 Full complement of pink  Normally 2 to 5 lobes
to rose-violet specific connected by thin
granules nuclear filament
 Abundant amount  Coarse, clumped
chromatin pattern
 Few nonspecific granules
are present
 NEUTROPHIL GRANULES
PRIMARY (AZUROPHILIC) SECONDARY TERTIARY SECRETORY
GRANULES (SPECIFIC) GRANULES GRANULES
GRANULES (SECRETORY
VESICLES)
• Myeloperoxidase • ß2-Microglobulin • Gelatinase • CD11b/CD18
• Acid ß-glycerophosphate • Collagenase • Collagenase • ALP
• Cathepsins • Gelatinase • Lysozyme • Vesicle-associated
• Defensins • Lactoferrin • Acetyltransferasae membrane-2
• Elastase • Neutrophil gelatinase- • ß2-Microglobulin • CD10, CD13, CD14,
• Proteinase 3 associated lipocalin CD16
• Transcobalamin I • Cytochrome b558
• Complement
receptor-1
NEUTROPHIL FUNCTIONS

 Innate Immune System

 Major function is PHAGOCYTOSIS and destruction
foreign material and microorganisms.
 Seeking
 Destruction

 Rolling, Adhesion, Crawling and Transmigration

 Phagocytosis
 Generation of NETs (NETosis)
 Secretion of Transcobalamin I or R binder protein
MATURE CELL - EOSINOPHIL

Size: 9-15 um in diameter

Cytoplasm
 Full complement of large, reddish-orange specific
granules

Nucleus
 Usually has 2 lobes
 Coarse, clumped chromatin pattern
MATURE CELL – BASOPHIL AND MAST CELLS
Size: 10-16 um in diameter
Cytoplasm
 Stains slightly pink to colorless
 Contains specific dark-purple to blue-black granules
 Granules are water soluble and tend to wash out when
stained

Nucleus
 Does not appear as coarse as in the neutrophil or
eosinophil
 Generally unsegmented or bilobed
Neutrophil Specific Eosinophil Specific Basophil Specific Granules
Granules Granules
•Lysozyme Larger Granules: •Histamine
•Lactoferrin •Major Basic Protein (MBP) •Heparin
•Collagenase •Phospolipase •Eosinophulic chemotactic factor-A
•Plasminogen activator •Cathepsin •Peroxidase
•Aminopeptidase •Eosinophil Cationic Protein
•Eosinophil derived neurotoxin
•Eosinophil protein X

Smaller Granules:
•Arylsulfatase
•Peroxidase
•Acid phosphatase
MONOPOIESIS
MONOCYTE FUNCTION

 Monocytes can also have a marginal and circulating pool

 They are able to survive for 30 hours in tissues
 Different patterns of chemokine receptors leads different tissue
destinations for monocytes
 Depending on the environment on the tissue, the monocytes
may become a macrophage, dendritic cell, or an osteoclast
MONOCYTE FUNCTION
 Innate immunity (Phagocytosis, cytokine production)

 Adaptive immunity (As APCs)

 Housekeeping
 Removal of debris at sites of infection
 Destruction of RBCs
 Protein synthesis
MONOBLAST
Size: 12-20 um in diameter
Cytoplasm
 Moderately basophilic to blue-gray
 Nongranular
Nucleus
 Ovoid or round in shape
 Light blue purple in color
 Fine, lacey chromatin
 1 to 2 nucleoli
 NC ratio is 4:1 to 3:1
PROMONOCYTE
Size: 14-18 um in diameter  NC ratio is 3:1 to 2:1
Cytoplasm
 Blue-gray
 May contain fine dustlike
azurophilic granules
 Groundglass appearance
Nucleus
 Oval, may have single fold
or fissure
 1 to 5 nucleoli
 Fine chromatin pattern
MONOCYTE
Size: 14-20 um in diameter Nucleus
Cytoplasm  Round, kidney shaped or
may show slight
 Blue-gray
lobulation (brainlike
 Outline may be irregular convulotion)
because of the presence
 No nucleoli
of pseudopods
 Chromatin is fine and
 Many fine azurophilic
lacey pattern
granules
 Ground glass appearance
 Vacuoles may be present
 MONOCYTE/MACROPHAGE FUNCTIONS
Innate Immunity Adaptive Immunity Housekeeping

• Toll-like receptors (recognize • Macrophages and Dendritic • Removal of debris and dead cells at
bacterial pathogens) cells site of infection or tissue damage,
• Synthesize Nitric oxide • Interact with T and B destruction of senescent RBCs and
maintenance of a storage pool of
lymphocytes
iron for erythropoiesis
MONOCYTES IN TISSUES
Liver Kuppfer cells
Lungs Alveolar macrophages
Brain Microglia
Skin Langerhans cells
Spleen Littoral cells
Intestines Intestinal macrophages
Peritoneum Peritoneal macrophages
Bone Osteoclasts
Lymph nodes Dendritic cells
Synovium Type A cell
Kidneys Renal macrophages
Reproductive organ macrophages
LYMPHOPOIESIS
 B Cells T Cells NK Cells
• Develop in BM as: pro-B, pre-B and • Develop in the Thymus as: pro-T, • Large granular lymphocytes
immature B cells pre-T and immature T cells • CD56, CD16, CD3 and CD7
• Immature B cells are also known as • Subdivided into 2 major categories: • Part of the innate immune system
Hematogones expression CD4 and CD8 • Kills virus-infected cells and tumor
• Effector B cells are antibody- • Mature T cells migrate to cells
producing cells known as plasma secondary lymphoid organs or in
cells and plasmacytoid cells the circulating blood
• CD4:
• TH1 – intracellular pathognes
• TH2 – extracellular parasites
(helminths), asthma
• TH17- extracellular bacteria and
fungi
• CD8+(effector lymphocytes) – kills
target cells, also known as cytotoxic
T lymphocytes
LYMPHOBLAST
Size: 10-18 um in diameter Nucleus
Cytoplasm  Chromatin pattern is
somewhat coarse
 Nongranular
 Round or oval in shape
 Appears smooth
 1 to 2 distinct nucleoli
 Moderate to dark blue
 NC ratio is 4:1
PROLYMPHOCYTE
Size: may be the same size as indented
the lymphoblast or smaller
 Chromatin pattern is
Cytoplasm more clumped
 Moderate to dark blue  May contain 1 to 2
nucleoli
 Usually nongranular, but
may contain occasional
azurophilic granules

Nucleus
 Round, oval or slightly
LYMPHOCYTE

 Small (8 to 10 µm dia.)
 Med (10 to 12 µm dia.)
 Large (12 to 16 µm dia.)
 Blue cytoplasm
 Round or oval nucleus
 No nucleoli are visible
T AND B LYMPHOCYTES
 Divided in 3 major groups: T cells, B cells and NK cells.
 T and B cells are major players in Adaptive Immunity.
 Specific
 Produces memory for reexposure
 Tolerance (self-antigens are ignored)

 Humoral response: producing antibodies

 Cellular response: attacking foreign organisms or cells
T AND B LYMPHOCYTES

 Lymphocytes are different from the other leukocytes in several ways, including the following:

1. Lymphocytes are not end cells. They are resting cells, and when stimulated, they undergo
mitosis to produce both memory and effector cells.
2. Unlike other leukocytes, lymphocytes recirculate from the blood to the tissues and back to
the blood.
3. B and T lymphocytes are capable of rearranging antigen receptor gene segments to produce
a wide variety of antibodies and surface receptors.
4. Although early lymphocyte progenitors such as the common lymphoid progenitor originate
in the bone marrow, T and NK lymphocytes develop and mature outside the bone marrow.
MEGAKARYOPOIESIS
MEGAKARYOBLAST
Size: 20-50 um in diameter kidney shaped
Cytoplasm  Fine chromatin pattern
 Varying shades of blue  Multiple nucleoli that
generally stain blue
 Usually darker than the
myeloblast  NC ratio is about 10:1
 May have small, blunt
pseudopods
 Nongranular

Nucleus
 Round, oval or may be
PROMEGAKARYOCYTE
Size: 20-60 um in diameter Nucleus
Cytoplasm  Chromatin becomes
more coarse
 More abundant than
previous stage  Multiple nucleoli are
visible
 Less basophilic than the
blast  Irregular in shape; may
even show slight
 Granules begin to form
lobulation
in the Golgi region
 NC ratio is 4:1 to 7:1
GRANULAR MEGAKARYOCYTE
Size: 30-90 um in diameter  Small in comparison to
cell size
Cytoplasm
 Multiple nuclei may be
 Pinkish blue in color
visible
 Very fine and diffusely
 Chromatin is coarser
granular
than the previous stage
 Usually has an irregular
 No nucleoli visible
peripheral border
 NC ratio is 2:1 to 1:1

Nucleus
MATURE MEGAKARYOCYTE
Size: 40-120 um in diameter  Multiple nuclei are
present
Cytoplasm
 No nucleoli visible
 Contains coarse clumps
of granules aggregating  NC ratio is less than 1:1
into little bundles, which
bud off from the
periphery to become
platelets.

Nucleus
PLATELET

Size: 1-4 um in diameter

Cytoplasm
 Light blue to purple
 Very granular

Nucleus
 none
Maturation stage Cytoplasmic Nuclear features Thrombocytes
granules visible
Megakaryoblast Absent Single nucleus, fine No
chromatin, nucleoli

Promegakaryocyte Few Double nuclei No

Megakaryocyte Numerous Two or more nuclei No or Few

Metamegakaryocyte Aggregated Four or more nuclei Yes

 PLATELET FUNCTION

 Thrombopoietin (TPO) is the main cytokine for thrombocytopoiesis

 The kidney is the primary source although it may also be elaborated in other organs such as the liver
 Platelets function to prevent loss of blood during vessel injury by forming platelet-plugs
 Adhesion
 Aggregation

 Other coagulatory substances help to stabilize the plug formed

 In cases where there is lack of platelets, Reticulated or Stress platelets are released which are larger than
usual platelets.
 Platelets also release Alpha & Dense Granules, which helps in the coagulation process.