Thalassaemia:

Pathogenesis and Lab Diagnosis

Dr. M Sadequel Islam Talukder
MBBS, M Phil (Pathology), MACP
Assistant Professor
Department of Pathology
Dinajpur Medical College
Presentation at Seminar on observation of World Thalassaemia Day 2009
Presentation: 10 May 2009, Dinajpur Medical College

Pathogenesis
Genetically determined

Heterogenous group of disorder

Reduced synthesis of one or more types of
normal haemoglobin polypeptide chain
Reduced haemoglobin involving affected chain

Normal Haemoglobin
HbA - 22
HbA2 - 22
HbF 22

Each goblin chain have separate genetic control

thalassaemia affect -chain synthesis

thalassaemia affect -chain synthesis

-Thalassaemia
An absence or deficiency of -chain synthesis of adult HbA

Chain synthesis

Hb-A

and chain
Hb-A = 22

On the basis of synthetic ability -genes are

designated as
gene can synthesize normal amount of -chain
+ gene can synthesize reduced amount of -chain
0 gene cannot synthesize -chain

 -thalassaemia major
Mutation of normal -gene 0-gene absence
HbA increased HA2 and HbF
genotype 00

-thalassaemia intermedia

HbA2
HbF
HbA
Genotype + + or 0

-thalassaemia minor
HbA2
HbA normal
HbF normal

Pathophysiology of -Thalassaemia
Various mutation in -gene

Complete or partial absence of -chain

Decreased adult HbA
-chain synthesis remain normal
Free complementary -chain unstable and
precipitate within normoblasts as insoluble inclusions
Cell membrane damage & impaired DNA synthesis
apoptosis i.e. ineffective erythropoeisis

70-80% marrow normoblasts undergo apoptosis

Inclusion bearing red cells undergo

sequestration & destruction in spleen

Partial or lack of HbA synthesis MCHC & MCH

Hypochromia & microcytosis

Normal

Thalassaemia

Reticulocytes undergo intramedullary death

Inadequate production + ineffective
erythropoiesis + haemolysis Anaemia

Haemolysis demands of phagocytic

function hyperplasia of phagocytes
Hepatosplenomegaly
To compensate anaemia extramedullary
haemopoiesis in liver, spleen & brain
Organomegaly

Erythropoiesis marrow expansion & thinning

of cortex of skull bone Thalassaemia facies

-Thassaemia
An absence or deficiency of -chain synthesis
due to delation of -genes.

Pathogenesis of -Thalassaemia
In normal individual HbA, HbA2 and HbF
need -chain for their formation.
4 genes of -chain, each pair on short arm of
chromosome 16 present with genotype
,/,.
In -thalassaemia, delation of -genes
reduction or absence of synthesis of -chain
depending on number of -gene delation.

 -chain synthesis free -chain in the fetus & chain in infant of 6 months, and continue in the
rest of life.
Complementary 4 and 4 are aggregated
Hb Bart (4 ) and HbH (4 ), respectively.

Variants of -Thalassaemia
Silent carrier

Delation of single -gene

Genotype /
Asymptomatic
Absence of RBC abnormality

Thalasaemia trait

Delation of 2 -genes
Genotype --/
Asymptometic, minimal or no anaemia
Minimal RBC abnormalities

 Hb H disease

Delation of 3 -genes
Genotype --/-
75% reduction of -chain
25% -chain synthesis small amount of HbF, HbA,
& HbA2
Fetus can survive
Severe anaemia
Severe RBC abnormalites

Hydrops fetalis

Delation of all -genes

Genotype --/-Absence of -chain synthsis
Only Hb Bart (4) is produced (High affinity for O2
and can not dissociate O2 to tissue)

Inheritance

Laboratory Diagnosis of Thalassaemia

Laboratory Findings

Hb concentration Decreased
ESR Mild increased
WBC Neutrophilic leucocytosis or normal
RBC count Markedly decreased
PCV Markedly decreased
MCV, MCH, MCHC reduced
Reticulocyte count Increased
Platelet count May be increased

Peripheral Blood Film

Normal

Morphology of PBF
RBC
Marked anisocytosis,
polikilocytosis,
microcytosis,
frequent target cells,
basophilic stippling,
fragmented cells or
schistocytosis,
polychromatic
macrocytes and
nucleated cells

WBC
Whin normal or
neutrophilic
leucocytosis

Platelets
Normal or increased

Thalassaemia slides

Haemoglobin Electrophoresis
Different types of haemoglobins contain
different surface charge which determine the
elecrtrophoretic mobility and gives a specific
bands on the electrophoretic papers when
haemolysate is undergone eletrophoresis.
HbF, HbA, HbA2, HbC, HbE, HbD, HbS, HbH
and Hb Bart can be measured accurately by
electrophoresis

Thanks