RBC Production and Destruction

Joseph Israel R. Guanlao, RMT, MSMT

Outline
I. Introduction
II. RBC Maturation Series
III. Characteristics of Erythroid Precursors
IV. Hypoxia
V. Erythropoietin
VI. RBC Destruction
Overview of Hematopoiesis
Introduction
 Red blood cells (aka Erythrocytes)
 Three nomenclature for naming the erythroblast
precursors:
ERYTHROBLAST

NORMOBLAST

RUBRIBLAST
RBC Maturation Series
ERYTHROBLAST NORMOBLAST RUBRIBLAST

Proerythroblast Pronormoblast Rubriblast

Basophilic erythroblast Basophilic normoblast Prorubricyte

Polychromatic Polychromatic
Rubricyte
erythroblast normoblast
Orthochromatic Orthochromatic
Metarubricyte
erythrolast normoblast
Polychromatophilic Polychromatophilic Polychromatophilic
erythrocyte erythrocyte Erythrocyte

Erythrocyte Erythrocyte Erythrocyte

Erythroid Progenitors
 BFU-E and CFU-E

 It takes 1 week for BFU-E to mature into CFU-E

and another week for CFU-E to mature into
pronormoblast

 It takes another 6 days for the precursors to

become mature RBCs
Erythroid Precursors

Basophilic Polychromatic
Pronormoblast
normoblast normoblast

Orthochromatic Polychromatophilic
ERYTHROCYTE
normoblast erythrocyte
Pronormoblast
 High N:C ratio

 Round to oval nucleus

 1-2 nucleoli

 Chromatin is open

 Blue cytoplasm (due to

high concentration of
ribosomes)
Pronormoblast
 Capable of division

 Present only in the

bone marrow

 Begins to accumulate
components for
hemoglobin
production
PRONORMOBLAST
Basophilic Normoblast
 Chromatin begins to
condense

 N:C ratio decreases

(6:1)

 Nucleoli may be
present but
disappears later
Basophilic Normoblast
 Cytoplasm appears
deeper blue than
pronormoblast

 Capable of division

 Present only in bone

marrow

 Detectable hemoglobin
synthesis occurs (but is
completely masked by
large number of
ribosomes and RNA)
BASOPHILIC NORMOBLAST
Polychromatic Normoblast
 More reduced N:C
ratio (4:1)

 Nucleoli are no longer

present

 Stained hemoglobin
can be seen
Polychromatic Normoblast
 Cytoplasm appears
pink and blue (murky
gray-blue)

 Last stage capable

of division

 Present only in the

bone marrow
POLYCHROMATIC NORMOBLAST
Orthochromatic Normoblast
 Completely
condensed nucleus

 Cytoplasm appears
pink-orange

 Not capable of
division due to
chromatin
condensation
Orthochromatic Normoblast
 Nucleus is ejected

 Howell-Jolly bodies
may be seen
ORTHOCHROMATIC
NORMOBLAST
Polychromatophilic Erythrocyte
 No nucleus

 Cytoplasm appears
similar with
orthochromatic
normoblast

 Remains larger than a

mature RBC
Polychromatophilic Erythrocyte
 Stays 1 day or longer
in the bone marrow
and another 1 day in
the circulation

 Retained in the
spleen for several
days for pitting and
polishing
Polychromatophilic Erythrocyte
 Completes the
production of
hemoglobin from
residual mRNA using
the remaining
ribosomes

 Referred to as
reticulocyte when
stained with supravital
stains
POLYCHROMATIC ERYTHROCYTE
Erythrocyte
 Biconcave disc
shape (enables
optimal gas
exchange)
 7-8 μm in diameter
with thickness of 1.5-
2.5 μm
 Appears salmon pink
with central pale
area
Erythrocyte
 Lifespan: 120 days

 Aging leads to their

removal in the spleen

 Membrane is flexible
and deformable that
allows them to
squeeze through
small spaces
Hypoxia
 Stimulus for RBC production

 Detected by peritubular interstitial cells of the

kidneys, which produces erythropoietin

 Production of EPO is increased when the oxygen-

carrying capacity of blood is diminished
Erythropoietin
 Thermostable, nondialyzable glycoprotein
hormone with a molecular weight of 34 kD

 Consists of a carbohydrate unit that reacts with

RBC receptors and terminal sialic acid unit.

 Mechanism of action:
1. Early release of reticulocytes
2. Inhibition of apoptosis
3. Reduced bone marrow transit time
Early release of reticulocytes
 RBCs in bone marrow is held by their surface
receptors for adhesion molecules

 EPO down-regulates the production of these

surface receptors on RBC surface

 Results to the presence of very basophilic

reticulocytes (shift/stress reticulocytes) and
even nucleated RBCs in cases of extreme
anemia
Early release of reticulocytes
 Short-term response (limited in effectiveness)
because precursors in bone marrow are depleted
within several days
Inhibition of Apoptosis
 Increases the number of cells that will be able to
mature into RBCs by decreasing apoptosis

 Process of Apoptosis:
1. Condensation of nucleus
2. Nucleolar disintegration
3. Shrinkage of cell volume
Inhibition of Apoptosis
 Fas  death receptor on early RBC precursors

 FasL  the ligand expressed by more mature

RBC precursors

 Fas-FasL cross-linking induces apoptosis of the

more immature RBC precursor

 EPO stimulates the early release of FasL-bearing

precursors
Inhibition of Apoptosis
 EPO induces the production of anti-apoptotic
molecules on the more immature erythroid
precursors
Reduced BM transit time
 EPO increases the rate of cellular processes and
decreases the cell cycle time (20% reduction)

 Also increases the rate of hemoglobin synthesis

Other hormones that influence
erythropoiesis…
 Testosterone – directly stimulates erythropoiesis

 Pituitary and thyroid hormones – affects the

production of erythropoietin
Erythrocyte Destruction
1. INTRAVASCULAR HEMOLYSIS

2. EXTRAVASCULAR HEMOLYSIS
Intravascular Hemolysis
 Mechanical or traumatic causes

 Cell contents are released in the surrounding

plasma and certain proteins (i.e. haptoglobin and
hemopexin) salvage hemoglobin so as to
preserve iron
Extravascular Hemolysis
 Occurs in the spleen
 Spleen
 Largest lymphoid organ
of the body
 Contains approximately
350 mL of blood
 Functions as an
indiscriminate filter of
circulating blood
 Composed of white
pulp, red pulp,
marginal zone
Extravascular Hemolysis
 Spleen (continued…)
 White pulp – consists
of scattered follicles
with germinal centers
containing
lymphocytes,
macrophages, and
dentritic cells
Extravascular Hemolysis
 Spleen (continued…)
 Marginal zone –
surrounds the white
pulp and forms a
reticular meshwork
containing blood
vessels and B cells
Extravascular Hemolysis
 Spleen (continued…)
 Red pulp –
composed of vascular
sinusoids and sinuses
separated by cords of
tissues containing
specialized
macrophages that
functions as a filter for
blood passing through
the region
Extravascular Hemolysis
Senescent Spleen (Red Sluggish
RBC Pulp) flow of blood

Plasma
pH Glycolysis
glucose
decreases slows down
level drops

RBC struggles to maintain iron in

Iron
reduced state (energy-dependent
oxidation
process)
Extravascular Hemolysis
 Electrolyte shifting (Na-K) causes senescent
RBCs to swell and become spherical
(spherocytes)

 Spherocytes are trapped in the splenic sieve

and are readily phagocytosed by splenic
macrophages

 Senescent RBCs succumb to the various

stresses present in the red pulp of the spleen
Extravascular Hemolysis
 2 methods of
removing senescent
RBCs in the spleen:
1. Culling – cells are
phagocytosed
2. Pitting - removal of
inclusions or
damaged surface
membrane from the
RBC
Extravascular Hemolysis
 After phagocytosis, Iron is removed from heme
and stored as ferritin in macrophages

 Globin portion of hemoglobin is returned to the

amino acid pool

 Protoporphyrin of heme is converted to bilirubin.