You are on page 1of 71

Prof Ammu K.

Radhakrishnan
Pathology Division,
School of Medicine, IMU
Learning Outcomes:
You will be able to:
1. Describe the production of
platelets and leucocytes
2. Name the major growth factors
involved in the production of
platelets and leucocytes

20 November, 2017 Prof Ammu K. Radhakrishnan 2


Haemopoiesis
1. Erythropoiesis
2. Megakaryopoiesis
3. Leucopoiesis

20 November, 2017 Prof Ammu K. Radhakrishnan 3


Megakaryopoiesis
Formation of platelets
Development & Maturation
Occurs in bone marrow

20 November, 2017 Prof Ammu K. Radhakrishnan 4


20 November, 2017 Prof Ammu K. Radhakrishnan 5
Platelets: Stages of
Development and Maturation
1. Haemopoietic stem cell
2. Myeloid stem cell
3. Megakaryoblast
4. Pro-megakaryocyte
5. Megakaryocyte
6. Platelets FOUND IN BLOOD

20 November, 2017 Prof Ammu K. Radhakrishnan 6


Megakaryocyte
Initially a mononuclear cell
Matures by endomitotic
synchronous nuclear replication
DNA replication without cytoplasmic
separation
No daughter cells produced

20 November, 2017 Prof Ammu K. Radhakrishnan 7


Megakaryocyte
Number of nuclear lobes
increases in multiples of two
There is no cell division
Polyploid cell
Cytoplasmic volumes enlarges
with each cell division
A large cell
50-150 m
20 November, 2017 Prof Ammu K. Radhakrishnan 8
Bone Marrow
Smear

20 November, 2017 Prof Ammu K. Radhakrishnan 9


Megakaryocyte
16th - 64th nucleus stage
nuclear replication and cell growth
stops
Microvesicles form
Platelets liberated
Enter marrow sinuses and blood
1 megakaryocyte
4000 platelets
20 November, 2017 Prof Ammu K. Radhakrishnan 10
Platelets

20 November, 2017 Prof Ammu K. Radhakrishnan 11


Platelet Production: Control

Thrombopoietin (TPO)
IL-6
IL-3
MK-CSF
Megakaryocyte-colony stimulating
factor

20 November, 2017 Prof Ammu K. Radhakrishnan 12


Thrombopoietin (TPO)
Megakaryocyte growth and
development factor
MGDF
353 amino acid protein
Encoded by the THPO gene
chromosome 3p27
High homology with erythropoietin
20 November, 2017 Prof Ammu K. Radhakrishnan 13
Thrombopoietin (TPO)
Produced by
Liver (main source)
Kidneys, testes, brain, bone marrow
stromal cells
Provide primary signals for
megakaryocyte production
Controls platelet production

20 November, 2017 Prof Ammu K. Radhakrishnan 14


Interleukin-6 (IL-6)
Similar function with TPO
Essential for forming adequate
quantity of platelets
Controls platelet production
Provide primary signals for
megakaryocyte production

20 November, 2017 Prof Ammu K. Radhakrishnan 15


IL-3 & MK-CSF
Exhibit megakaryocyte stimulating
activity
Promote formation of
megakaryocyte colony

20 November, 2017 Prof Ammu K. Radhakrishnan 16


20 November, 2017 Prof Ammu K. Radhakrishnan 17
Leucopoiesis
Production of leucocytes
White blood cells
Account for <1% of whole blood
Crucial for immune system
Protects against infections
bacteria, viruses, parasites etc.

20 November, 2017 Prof Ammu K. Radhakrishnan 18


Granulocytes

20 November, 2017 Prof Ammu K. Radhakrishnan 19


Agranulocytes

10-14 mm

T-, B-lymphocytes
Natural Killer (NK) cell

20 November, 2017 Prof Ammu K. Radhakrishnan 20


Leucopoiesis
Sites:
Bone marrow (main site)
Thymus
3 major processes:
1. Granulocytopoiesis
2. Monocyte development
3. Lymphopoiesis

20 November, 2017 Prof Ammu K. Radhakrishnan 21


Leucopoiesis
All leucocytes originate from the
haemopoietic stem cells
Leucocytes can arise from
1. Myeloid stem cells
2. Lymphoid stem cells

20 November, 2017 Prof Ammu K. Radhakrishnan 22


Haemopoietic Stem Cell

Myeloid Stem Cells Lymphoid Stem Cells

Megakaryocyte GM Progenitor
Progenitor

Proerythroblast Myeloblast Monoblast

Platelets
Erythrocytes Monocyte/
Granulocytes
Prof Ammu K. Radhakrishnan
Macrophage23
20 November, 2017
Haemopoietic Stem Cell

Myeloid Stem Cells Lymphoid Stem Cells

B-lymphocytes Progenitor
Progenitor
T-lymphocytes NK
Progenitor Progenitor
B-Lymphocytes

20 November, 2017
T-Lymphocytes
Prof Ammu K. Radhakrishnan
NK cells24
Terminology
IL
CFU
Interleukin
Colony forming
units SCF
BFU Stem cell factor
Burst forming CSF
units Colony
stimulating
factor

20 November, 2017 Prof Ammu K. Radhakrishnan 25


Granulocytes: Development
Originate from HSC in the red
bone marrow
Maturation driven by:
Granulocyte-monocyte colony
stimulating factor (GM-CSF)
Granulocyte colony stimulating
factor (G-CSF)

20 November, 2017 Prof Ammu K. Radhakrishnan 26


Haemopoiesis: Growth Factors and
Development
SCF
SCF
IL-3
Flt-3L
Myeloid IL-6
Stem Cells

Haemopoietic
Stem Cell

SCF
IL-3
Lymphoid
Stem Cells
IL-6
20 November, 2017 Prof Ammu K. Radhakrishnan IL-7 27
Granulocytopoiesis: Growth Factors
and Development
Myeloid
Stem Cells
GM Progenitor
GM-CSF cells

Haemopoietic
Stem Cell Other Progenitor
cells

Lymphoid
Stem Cells

20 November, 2017 Prof Ammu K. Radhakrishnan 28


Granulocytopoiesis: Growth Factors
and Development

G-CSF
GM-CSF

Myeloid GM Myeloblast
Stem Cells Progenitor
cell

20 November, 2017 Prof Ammu K. Radhakrishnan 29


Granulocytopoiesis: Growth
Factors and Development

**
Myeloblast Promyelocyte Myelocyte

20 November, 2017 Prof Ammu K. Radhakrishnan 30


Myelocyte
Precursor/progenitor for all
granulocytes
One for each of the granulocytes
Neutrophil myelocyte
Eosinophil myelocyte
Basophil myelocyte

20 November, 2017 Prof Ammu K. Radhakrishnan 31


Granulocytopoiesis: Growth
Factors and Development

Myelocyte Metamyelocyte Band Cell

Mature
Granulocyte
20 November, 2017 Prof Ammu K. Radhakrishnan 32
Granulocytopoiesis
2 steps:
1. Proliferation/Mitotic
2. Differentiation & maturation

20 November, 2017 Prof Ammu K. Radhakrishnan 33


Granulocytopoiesis:
Proliferation
Rapid mitotic division
Intermediate stages involved:
Myeloblast
Promyelocyte
Myelocyte
Duration
7.5 days

20 November, 2017 Prof Ammu K. Radhakrishnan 34


Granulocytopoiesis:
Differentiation & Maturation
Post mitosis
Cells undergo maturation and
differentiation to develop characteristic
morphology and feature
Stages involved
Metamyelocyte
Band cell
Duration
6.5 days
20 November, 2017 Prof Ammu K. Radhakrishnan 35
Neutrophils: Growth Factors and
Development

Myeloblast
Promyelocyte

GM Neutrophil
Myeloid
Progenitor Myelocyte
Stem Cells
cell

Metamyleocyte
Band cell

NEUTROPHIL
G-CSF, GM-CSF
20 November, 2017 Prof Ammu K. Radhakrishnan 36
Eosinophils: Growth Factors and
Development

Myeloblast
Promyelocyte

GM Eosinophil
Myeloid
Progenitor Myelocyte
Stem Cells
cell

Metamyelocyte
Band cell

EOSINOPHIL
IL-3, IL-5
20 November, 2017 Prof Ammu K. Radhakrishnan 37
Basophils: Growth Factors and
Development

Myeloblast
Promyelocyte

GM Basophil
Myeloid
Progenitor Myelocyte
Stem Cells
cell

Metamyelocyte
Band cell

BASOPHIL
?
20 November, 2017 Prof Ammu K. Radhakrishnan 38
Granulocytes: Stages seen
only in bone marrow
1. Haemopoietic stem cells
2. Myeloid stem cells
3. Myeloblasts
4. Promyelocytes
5. Myelocytes
6. Metamyelocytes

20 November, 2017 Prof Ammu K. Radhakrishnan 39


Granulocytes: Stages seen in
the periphery
1. Blood
Band cells
Mature granulocytes
2. Connective tissues
Mast cells

20 November, 2017 Prof Ammu K. Radhakrishnan 40


Peripheral Blood

20 November, 2017 Prof Ammu K. Radhakrishnan 41


Monocytes: Development
Originate from haemopoietic stem
cell in the red bone marrow
Arise from the myeloid stem cell
Maturation driven by:
GM-CSF
Monocyte-colony stimulating factor
(M-CSF)

20 November, 2017 Prof Ammu K. Radhakrishnan 42


Haemopoiesis: Growth Factors and
Development
Myeloid
Stem Cells
GM Progenitor
GM-CSF cells

Haemopoietic
Stem Cell Other Progenitor
cells

Lymphoid
Stem Cells

20 November, 2017 Prof Ammu K. Radhakrishnan 43


Monocytes: Growth Factors and
Development

M-CSF
Myeloid GM Monoblast
GM-CSF
Stem Cells Progenitor
cell

Promonocyte
20 November, 2017 Prof Ammu K. Radhakrishnan 44
20 November, 2017 Prof Ammu K. Radhakrishnan 45
Monocytes: Forms only seen
in bone marrow
1. Haemopoietic stem cells
2. Myeloid stem cells
3. Monoblasts
4. Promonocytes

20 November, 2017 Prof Ammu K. Radhakrishnan 46


Monocytes: Forms seen in
the periphery
1. Blood
Monocytes
2. Tissues
Macrophages
Various types

20 November, 2017 Prof Ammu K. Radhakrishnan 47


Lymphopoiesis
Types of cell
1. T-lymphocytes
TCR ab T-cells
TCR gd T-cells
2. B-lymphocytes
3. NK cell

20 November, 2017 Prof Ammu K. Radhakrishnan 48


Lymphopoiesis

Adapted from Ref (3)


20 November, 2017 Prof Ammu K. Radhakrishnan 49
Lymphocytes: Development

Takes place in:


1. Bone marrow
B-lymphocytes & NK cells
2. Thymus
T-lymphocytes
Maturation driven by cytokines
IL-2, IL-4, IL-6, IL-7
20 November, 2017 Prof Ammu K. Radhakrishnan 50
Lymphocyte Development

Pluripotent haemopoietic
Lymphoid stem cell
stem cell

Lymphoid
Progenitor

B-lymphocyte
T-lymphocyte
Progenitor
NK Progenitor
(Pro B-cells)
Progenitor (Pro T-cells)
20 November, 2017 Prof Ammu K. Radhakrishnan 51
T-Lymphocyte: Development

Pro T-cells
CD4- CD8- TCR- CD3-
Double negative (DN)
Maintained by:
IL-6, IL-7

20 November, 2017 Prof Ammu K. Radhakrishnan 52


T-Lymphocyte: Development
Rearrangement
Migrate to thymus of TCR heavy
chain (b) genes
(Pro T-cells)

20 November, 2017 Prof Ammu K. Radhakrishnan 53


Immature T-lymphocyte
V, D, J genes rearrange
Form functional VDJ unit
Produce functional b-chain
A productive VDJ rearrangement
required for T-cell survival

20 November, 2017 Prof Ammu K. Radhakrishnan 54


Immature T-lymphocyte
b-heavy chain
Associates with a surrogate light
chain
Pre-TCR

Pre-TCR+
CD4-CD8- CD3-
20 November, 2017 Prof Ammu K. Radhakrishnan 55
T-Lymphocyte: Development
Rearrangement
of TCR heavy
Migrate to thymus
chain (b) genes
(Pro T-cells)

IL-2, IL-4

Pre-TCR+
CD4- CD8- CD3- Immature T-lymphocyte
20 November, 2017 Prof Ammu K. Radhakrishnan 56
Pre-TCR
Required for survival and
proliferation of immature T-cells
Regulates further rearrangement
TCR g genes
1. Stimulate light chain
rearrangements
2. Produce functional a-chain of
TCR
20 November, 2017 Prof Ammu K. Radhakrishnan 57
TCR
Adapted from
Ref (3)

20 November, 2017 Prof Ammu K. Radhakrishnan 58


TCR
Functional b- and a- chain
ab-TCR
Signals for expression of
CD4
CD8
CD3

20 November, 2017 Prof Ammu K. Radhakrishnan 59


T-Lymphocyte: Development

Immature T-lymphocyte
Pre-TCR+ CD4-CD8- CD3

CD3- TCRab+
CD4+ CD8+

20 November, 2017 Prof Ammu K. Radhakrishnan 60


T-Lymphocyte: Development

TCRab+ CD3+
CD3- TCRab+
CD4+ CD8+
CD4+ CD8+
DOUBLE POSITIVE (DP)
20 November, 2017 Prof Ammu K. Radhakrishnan 61
Double Positive T-
lymphocytes
Intermediate stage
Present in thymus
Absent in peripheral circulation
Express
ab-TCR, CD3
CD4, CD8
Undergo positive selection
20 November, 2017 Prof Ammu K. Radhakrishnan 62
Positive Selection
Double positive (DP) become
single positive (SP) cells
1. TCRab+ CD3+ CD4+ CD8-
2. TCRab+ CD3+ CD8+ CD4-
How?
Depends on affinity of TCR to MHC
proteins

20 November, 2017 Prof Ammu K. Radhakrishnan 63


20 November, 2017 Prof Ammu K. Radhakrishnan 64
gd-TCR T-lymphocytes
Function is not clear
Reported to play a critical role:
Immune regulation
Tumour surveillance
Some primary immune responses
Derived and mature from the same
pro-T cells that also generate the
ab-TCR T-cells
20 November, 2017 Prof Ammu K. Radhakrishnan 65
B-Lymphocyte Subsets

+ Lymph
nodes

16 June 2014 Prof Ammu K Radhakrishnan Adapted from Ref66(3)


Ig Expression During B-Cell
Development

Nave
B-cell

16 June 2014 Prof Ammu K Radhakrishnan


Adapted from Ref67 (3)
Checkpoints in Lymphocyte
Maturation

Adapted from Ref (3)


20 November, 2017 Prof Ammu K. Radhakrishnan 68
NK cell: Development
Originates from lymphoid stem
cell
Process occurs in bone marrow
Development stages not yet fully
elucidated

20 November, 2017 Prof Ammu K. Radhakrishnan 69


20 November, 2017 Prof Ammu K. Radhakrishnan 70
References
1. Abbas & Lichtman; Basic Immunology (4th Edition);
Elsevier Saunders
2. Peakman and Vergani; Basic and Clinical Immunology
(2nd Edition);
Churchill Livingstone
3. Abbas, Lichtman & Pillai; Cellular and Molecular
Immunology (7th Edition); Elsevier Saunders
4. Haematology an illustrated text (2nd edition). Howard
& Hamilton (Churchill Livingstone)
5. Essential Haematology (5th edition)
Hoffbrand and Pettit (Blackwell)

20 November, 2017 Prof Ammu K. Radhakrishnan 71