Haematopoiesis,

haemoglobin structure and

blood cell abnormalities
Learning aims

 The aims of this session is to give a theoretical grounding

framework that encompasses the following:
 Normal haematopoiesis
 Erythropoiesis

 Thrombopoiesis

 Leukopoiesis

 Haemoglobin function and variants

 Introduction to basic red cell pathology
Learning outcomes
 By the end of this session students will have:
 Hadthe basic cellular process of haematopoiesis
demonstrated
 Have seen light microphotographic images of all cell
lines
 Gain an insight into the hormonal controls used to
generate and regulate blood cells
 Have see light microphotographs of elementary red cell
pathologies
 Begin to converse in haematologica
The end is where we start from
The Cells

 All start in the bone marrow

 Neutrophils, monocytes, platelets and eosinophils are

MYELOID

 Lymphocytes are Lymphoid

Neutrophils

Morphology:
Purple, granular with 4-6
lobes
Role:
Innate immunity, effective
against bacteria and debris
Detection:
FBC, Blood Film, Flow
Cytometry
Monocytes

Morphology:
Lighter hue of purple,
mildly granular, very large
cell, mononuclear, have
vacuoles
Role:
Innate immunity, become
tissue macrophages,
effective against bacteria
and debris
Detection:
FBC, Blood Film, Flow
Cytometry
Eosinophils

Morphology:
Striking red hue, very
granular, bi or tri lobed
nucleus
Role:
Innate immunity, allergic
reactions, parasites
Detection:
FBC, Blood Film, Flow
Cytometry
Basophils

Morphology:
Striking blue hue, very
granular, nucleus almost
occluded by granules
Role:
Largely unknown
Detection:
FBC, Blood Film, Flow
Cytometry
Erythrocytes

Morphology:
A-nuclear bi-concave disc,
distinctive pink/red hue
Role:
Transport oxygen to the
tissue
Detection:
FBC, Blood Film, Flow
Cytometry
Sites of haematopoiesis

 myeloid tissue
 bone marrow
 erythrocytes
 platelets
 granulocytes
 monocytes
 lymphoid tissue
 thymus
 lymph nodes
 spleen
Erythropoiesis: genesis of red blood cells
(Fig 17.5 from: Marieb & Hoehn, 2007)
Model of the hematopoietic developmental hierarchy
(Bryder, Rossi & Weissman, 2006)
Development of Blood Cells

 3/52 : formation of blood

islands from yolk sac
 6/52 : liver becomes
hematopoietic organ
 6/52 : spleen (until 8th
month)
 12-14/52 : bone marrow
(life-long)
Figure (a) Bone marrow cells are increasingly differentiated and lose the capacity for self-renewal as they mature. (b) A single stem cell gives rise, after multiple cell
divisions (shown by vertical lines), to > 10 6 mature cells.
Hoffbrand, A. Victor; Moss, Paul A. H.. Hoffbrand's Essential Haematology (Essentials) (Kindle Location 1661). Wiley. Kindle Edition.
Haemopoiesis
 Haemopoiesis occurs in a
suitable microenvironment (‘
niche’) provided by a stromal
matrix on which stem cells grow
and divide.
 The niche may be vascular
(lined by endothelium) or
endosteal (lined by osteoblasts).
 Extracellular glycoproteins and
other compounds are involved in
the binding.

Hoffbrand, A. Victor; Moss, Paul A. H.. Hoffbrand's Essential Haematology

(Essentials) (Kindle Locations 1670-1671). Wiley. Kindle Edition.
Haematopoiesis
Growth factors may stimulate
proliferation of early bone
marrow cells, direct
differentiation to one or other
cell type, stimulate cell
maturation, suppress apoptosis
or affect the function of mature
non-dividing cells, as illustrated
here for granulocyte colony-
stimulating factor (G-CSF) for an
early myeloid progenitor and a
neutrophil.
Hematopoiesis

Proliferative potential differentiation

The Role of Growth Factors
Granulocyte maturation
stem cell: <0.1%
myeloblasts: ~2%
promyelocytes: ~5%

myelocytes: 5-20%

metamyelocytes: ~22%

granular leucocytes
Haematopoietic Response

hypoxia RBC

infection granulocyte/monocyte

antigen lymphocyte

haemorrhage platelet
Erythropoiesis

 erythropoietin-independent stage:
 GM-CSF

 erythropoietin-dependent stage:
 erythropoietin
Granulopoiesis

 early phase: GM-CSF SCF IL-3

 Neutropoiesis: G-CSF
 Monopoiesis:
M-CSF
 Eosinopoiesis:
 Basopoiesis,Mastpoiesis:
IL-5 IL-3 GM-CSF
SCF IL-3
Lymphopoiesis

 B-cells:
 initial stage: IL-7 SCF
 later stage: Fcg rec IL-4 IL-6
 final proliferation and Ab secretion:
 T-cells: IL-6 GM-CSF
 CD8 cells: IL-2 Ag
 CD4 cells:
TCR/CD3 CD28
B-cell maturation
B-cell transformation
T-cell transformation
Markers

 Stem cell: CD34, c-kit

rhodamine, Hoechst dyes (pale)

 B-cell: CD19, CD20, (CD22), CD79a

 T-cell: CD3, CD2, CD5, CD4/CD8

 NK-cell: CD16, CD57, CD56

Red cell membrane: the basics

Three dimensional representation of a cell membrane 33

Red cell constituents

34
Red cell membrane constituents
continued….

35

Four major phospholipids in mammalian plasma membranes

The structure of the red cell membrane
Basic Structure

37
Haemoglobin synthesis in the
developing red cell. The
mitochondria are the main
sites of protoporphyrin
synthesis, iron (Fe) is
supplied from circulating
transferrin; globin chains are
synthesized on ribosomes. δ-
ALA, δ-aminolaevulinic acid;
CoA, coenzyme A.
Structure of Haem
The oxygenated and deoxygenated haemoglobin molecule. α, β, globin
chains of normal adult haemoglobin.
Other sickling haemoglobins (in addition to Hb S):
Hb SAntilles, Hb CZiquinchor, Hb CHarlem, Hb SProvidence, Hb SOman,
Hb STravis, Hb SSouth End, Hb SJamaica Plain, Hb SCameroon 41
(Lewis, Bain & Bates, 2006)
But not just for oxygen

Fig 7.22 From: Berg, Tymoczko & Stryer, 2006

42
 The Bohr Curve

A reduction in the total

binding capacity of
haemoglobin to oxygen
(i.e. shifting the curve
down, not just to the
right) due to reduced pH
is called the root effect.

PO2
Cooperative Binding of Haemoglobin
 Compensating mechanisms in anemia:

 The release of oxygen to the tissues is increased (reduced oxygen affinity

of Hb)
Red cell pathology
anisocytosis
poikilocytosis

anisocytosis(aniso = unequal)
 various sizes

poikilocytosis (poikilo = various)

 various shapes
elliptocytes

- heredirary
elliptocytosis

- iron def. anaemia

myelofibrosis
with
myeloid metaplasia

- megaloblastic anemia


spherocytes

- hereditary
spherocytosis

- acquired haemolytic
anaemia (e.g.
AIHA)

- physical or chemical
injury
- heat
leptocytes
(target cells)
- liver disease
 (obstructive jaundice)
- post splenectomy
- haemoglobinopathies
 (hypochromic anaemias)

thalassemia
Hgb C disease
Hgb H disease
schistocytes
(cell fragments)
indication of haemolysis

- megaloblastic anaemia

- severe burns

- traumatic haemolysis

-microangiopathic
haemolytic anemia
acanthocytes
(irregular surface spicules)
 irregularly spiculated
cells

 with bulbous/rounded
ends of spicules

liver disease
echinocytes
(crenated cells, burr cells)
regularlycontracted cells with
smooth surface undulation

- uremia

- artefact

- hyperosmolarity

- discocyte-echinocyte
transformation
Bite cells

Removal (“bites”) of
membrane by splenic
macrophages

- G6PD deficiency
dacrocytes
(teardrop cells)
- thalassemia

- myelofibrosis
drepanocytes
(sickle cells)
- sickle cell anemia
rouleaux

RBC stack like coins

- multiple myeloma
Howell-Jolly body
remnant of nuclear
chromatin

single:

•megaloblastic anemia
•hemolytic anemia
•post splenectomy
multiple:

•megaloblastic anemia
•otherabnormal
erythropoiesis
 Heinz bodies

denatured haemoglobin

- G6PD deficiency

Inclusions within red blood cells composed of denatured haemoglobin.

Conclusion

 All cells come from other cells

 All cells have a tightly controlled mechanism of

production

 Haemoglobin has many variants that affect its structure

 All red cell types have a name