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Erythropoiesis
Professor Amany Ahmed Osman
Professor of Hematology
Ain Shams University
ILOs of erythropoiesis:
• -to define the process of erythropoiesis, its stages & criteria of
maturation
• -to recognize reticulocytic counts & diagnostic significance
• -to list requirements of effective erythropoiesis
• -to discuss Hb synthesis
• -to know basic biochemical & physiological information regarding
RBCmetabolism, Hb structure & function
• -to list RBC abnormalities
• -to discuss ineffective erythropoiesis: causes & lab findings
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Erythropoiesis
• Def: the process of producing red blood cells by the stem cells in the
bone marrow
• Stem cell→CFUGEMM→BFUE→CFUE→Pronormoblast.
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CFU-E vs BFU-E
• CFU-E colony of erythroblasts containing 16 cells. • BFU-E (group of colonies) of erythroblasts containing
over 1,000 cells.
Pronormoblast:
• 1st recognizable eryrthroid precursor in BM.
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12-20 u
10-16 u
10-14 u
10-12 u
8.5 u
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Erythroid series
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Erythroid series
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Erythroid series
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Reticulocytes
•Reticulocytes are immature non-nucleated RBCs that contain remnants of
ribosomal RNA which continue to synthesize Hb after loss of their nucleus.
•Spends 1-2 days in BM & circulates in PB where complete loss of RNA occurs
mainly in the spleen. This maturation process takes approximately 2 – 3 days, of
which about 24 hours are spent in the circulation to give a completely pink-staining
mature non-nucleated biconcave disc.
•Reticulocytes appear in Leishman stained films larger than mature RBCs and show
polychromasia.
Normoblasts
• Normally, no normoblasts appear in adult PB.
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c)Reticulocytic production index: Still this correction is not enough as in cases of increased
demands, there is an increase in erythropoietic stimulus which leads to premature release in the
circulation and these stress reticulocytes have a maturation time as long as 3 days. So further
correction is needed:
reticulocytic count x Hct of patient/normal Hct(45%) x 1/maturation time in days(acc. to Hct
value)
45% →1 day, 35%→ 1.5 days, 25% →2 days, 15%→2.5 days. (Normal RPI: 2-3)
d) Absolute reticulocytic count: is calculated by multiplying the reticulocytic count x the RBC
count.
Normal range: Birth: 120-400 x 109/L
6-12 years: 30-100 x 109/L
Adults: 50-100 x 109/L
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Diagnostic significance:
1-The number of reticulocytes & percentage is a good indicator of bone marrow activity:
-Reticulocytosis (abnormally high numbers of reticulocytes):
When there is an increased production of red blood cells to overcome chronic or severe loss
of mature red blood cells, such as in a hemolytic anemia or hemorrhage.
-Reticulocytopenia(Abnormally low numbers of reticulocytes) can be attributed to
chemotherapy, aplastic anemia, or factors impairing normal reticulocytic response to
anemia as in: marrow disease (hypoplasia, infiltration by carcinoma, lymphoma, myeloma, AL,
TB), deficiency anemia (iron, folate, B12), lack of epo (renal disease), chronic disease,
ineffective erythropoiesis( thalassemia major, megaloblastic anemia, MDS, MF, CDA).
Diagnostic significance:
2-In cases of anemia, reticulocyte percentage should be higher than "normal" if the bone
marrow's ability to produce new blood cells remains intact. Calculating the reticulocytic
production index is an important step in understanding whether the reticulocyte count is
appropriate or inappropriate to the situation.
RPI < 2 indicates ineffective erythropoiesis.
3-Therapeutic trial test: reticulocytic counts can also be used to monitor the progress of
treatment for anemia where the earliest rise as a response can be recorded as early as 3
days post treatment and peaking after 7 days of therapy after recording base line
reticulocytes.
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Erythropoietin(Epo)
• Epo gene.
• A heavily glycosylated polypeptide (165 aa) of MW 34 kDa.
• Production sites:
-90% in peritubular interstitial kidney cells
-10% in liver & elsewhere.
There are no preformed stores.
• Stimulus for secretion: O2 tension in kidney tissues.
• Effect: ↑ no of progenitor cells committed to erythropoiesis →
erythroid hyperplasia in BM.
Epo Drive
• ↑ Epo: • ↓ Epo:
- Anemias -High red cell mass
- Abnormal Hb(struct or metab), -Some abnormal Hb which
unable to give up O2 easily release O2 more readily than
-Low atmospheric O2 normal
-Defective cardiac or pulmonary
function
-Damage to renal circulation
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Erythroid hyperplasia
• Transcription factors GATA-1 & FOG-1 are activated by Epo receptor
stimulation→ enhance expression of:
-erythroid-specific genes (heme biosynthesis & RBC membrane
proteins)
-anti-apoptotic genes
-transferrin receptor (CD71)
Late BFUs & CFUs (have Epo recpetors) are stimulated to proliferate,
differentiate &produce Hb.
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Hemoglobin
• Function:- it is a specialized protein in RBCs.
-in arterial blood, carries O2 from lung to tissues & in venous blood, carries
CO2 from tissues to lungs.
• Each RBC contains ~ 640 million Hb molecules
• MW of Hb is 68,000 dalton
• Each Hb molecule consists of 4 polypeptide chains, each binds to 4 O2 molecules
• Normal Hbs in adult blood
Hb A Hb F Hb A2
Structure α2β2 α2γ2 α2δ2
Normal(%) 96-98
CP6002
<Prof.1Amany Ahmed Osman 1.5-3.2
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• Iron metabolism
Background
• Hb is the primary vehicle for transporting oxygen in the blood.
• Oxygen is also carried dissolved in the blood's plasma, but to a lesser degree.
• Each Hb molecule has the capacity to carry four oxygen molecules. How much of that
capacity is filled by oxygen at any time is called the oxygen saturation. Expressed as a
percentage, the oxygen saturation is the ratio of the amount of oxygen bound to the Hb,
to the oxygen carrying capacity of the Hb.
• The oxygen carrying capacity is determined by the amount of Hb present in the blood.
• The amount of oxygen bound to the Hb at any time is related, in large part, to the partial
pressure of oxygen to which the Hb is exposed. In the lungs, at the alveolar-capillary
interface, the partial pressure of oxygen is typically high, and therefore the oxygen binds
readily to Hb that is present.
• As the blood circulates to other body tissue in which the partial pressure of oxygen is less,
the Hb releases the oxygen into the tissue because the Hb cannot maintain its full bound
capacity of oxygen in the presence of lower oxygen partial pressures.
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O2-Dissociation Curve
• The affinity of Hb to O2 is nicely balanced so that: -in lungs, Hb is 95-
96% oxygenated
-in tissues, Hb can give up O2 as much as possible
• Physiologically: -if affinity to O2 was less → complete oxygenation in
the lungs could not be reached
-if affinity to O2 was more → complete release of O2 to
tissues could not be reached
•P50: partial pressure of O2 at which Hb is ½ saturated with O2. N, P50= 26.6 mmHg
•It is a conventional measure of Hb affinity for oxygen.
•Normal position of curve depends on:
-2-3 DPG conc -H+ conc -CO2 conc -Hb molecule structure
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A lipid bilayer: integral membrane proteins & skeleton, 50% protein, 20% PL, 20% cholesterol & 10% CHO. CHO occur on outer
surface (ags) & proteins either peripheral or integral (penetrating the bilayer)
CP6002 Prof. Amany Ahmed Osman
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RBC abnormalities
RBC inclusions
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Ineffective erythropoiesis
• Def: inefficient erythropoiesis as 10-15% of developing erythroblasts
die within the marrow (intramedullary hemolysis) or other sites of
production and/or production of non viable RBCs which survive only
for a short time in the circulation.
Causes
• Extramedullary erythropoiesis : MF
• Hyperplastic but functionally abnormal BM:
-Thalassemia major & intermedia (free globin chains)
- Megaloblastic anemia (slowed DNA synthesis)
-IDA
-MDS
-Sideroblastic anemias
-Congenital dyserythropoietic anemias
-Erythroleukemias
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Anemia
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