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MLT 502 –
IMMUNOHAEMATOLOGY
II

HS221/5

GROUP C
HEMOLYTIC
DISEASE OF NEWBORN
(HDN)
DEFINITION
A condition in which fetus or neonate’s red
blood cell (RBC) are destroyed by
Immunoglobulin G (IgG) antibodies
produced by mother.
 "hemolytic" means breaking down of
red blood cells
 "erythroblastosis" refers to making
of immature red blood cells
 "fetalis" refers to fetus
HISTORY
Haemolytic disease of the newborn (HDN) has been known for
centuries with its typical and poor prognosis. The first
described by a French midwife who delivered twins in 1609.
One baby was swollen and died soon after birth, the other baby
developed jaundice and died several days later. For the next 300
years, many similar cases were described in which newborns
failed to survive.
During 1970s, the routine antenatal care included screening of
all expectant mothers to find those whose pregnancy may be at
risk of HDN, and giving preventative treatment accordingly.
This has led to decrease cases in the incidence of HDN,
particularly severe cases that were responsible for stillbirth and
neonatal death.
Hemolytic Disease of Newborn (HDN)
CAUSES OF HDN

Rh
INCOMPATIBILITY

ABO
INCOMPATIBILITY

OTHER SYSTEM
ANTIBODIES
INCOMPATIBILITY

OTHER Rh
ANTIBODIES

OTHER ANTIBODIES
CAUSES OF HDN
1) Rh incompatibility
- HDN is occured when a mother with Rh-negative blood
becomes pregnant with Rh-positive baby that inherited from
Rh-positive father.
- It occurs when anti-D is stimulated in mother plasma due to
mother ‘s immune response to the antigen D on fetal’s red
blood cells.
- This is due to anti-D is an IgG that capable to cross
placenta and hence delivered to fetal circulation.
- Rh caused HDN is less common but more severe.
2) ABO incompatibility
- HDN is arouse when a mother with blood type O becomes
pregnant with a fetus with different blood types A, B or AB.
- ABO antibodies is natural occurring antibodies that
clinically significant.
- ABO caused HDN is commonly occur but less severe.
3) Others unexpected immune antibody (other than anti-D) :
- Other Rh antibodies :
- anti-E (second most common, mild disease)
- anti–c (third most common, mild to severe)
- anti-C and anti-e (rare)
- antibody combination (anti-c and anti-E occurring
together, can be severe)
- Other antibodies
- Kell system antibodies (uncommon causes)
- Duffy, MNSs and Kidd system antibodies (rare
causes)
* Lewis and P system antibody are no occurrence in HDN.
Hemolytic Disease of Newborn (HDN)
PHYSIOLOGIC OF HDN
Physiologic of HDN is defined as
process of how HDN occur
(mechanism).
RHESUS HEMOLYTIC
DISEASE OF NEWBORN
• In a first pregnancy, Rh sensitization is not likely. Usually it
only becomes a problem in a future pregnancy with another
Rh positive baby.
• When the next pregnancy occur, the mother's antibodies
cross the placenta then reacts with an RBC antigen to fight
the Rh positive cells in the baby's body that the baby has
inherited from the father, and that is foreign to the mother.
• Hence, the antigen antibody interaction occurs.
• Sensitization of baby’s red blood cell (RBC) by mother’s IgG
antibody causes the baby’s RBC to be destroyed.
• These antibody-coated RBCs are removed from fetal circulation
by the macrophages of the spleen and liver.
• The severity of anemia depends on the amount of mother’s
antibody, its specificity, its avidity, and others characteristics.
• Anemia will stimulate bone marrow to produce more RBC
including immature RBC, which is then released to fetus
circulation. This is also known as erythroblastosis fetalis.
This figure shows pathophysiology of HDN
Retrieved from: http://tmedweb.tulane.edu/pharmwiki/lib/exe/fetch.php/rhod.png
This figure explains the Mechanism of HDN
Retrieved from: http://legacy.owensboro.kctcs.edu/gcaplan/anat2/notes/14_23.jpg
ABO INCOMPATIBILITY OF
HDN
• ABO HDN is due to incompatibility of ABO between
mother and fetus.
• A portion of the mother’s ABO antibodies may be IgG
and cross the placenta.
• It thus sensitizes the fetal cells.
• This condition can occur on the first pregnancy and can
occur without there having been transfusion.
• The mechanism of ABO HDN is similar to the RH HDN.
• Occurrence of ABO HDN is somewhat common, but
fortunately it is not often very severe.
This figure shows circulating IgG maternal antibodies cross the
placenta and sensitize fetal antigens positive cells.
Retrieved from Essential of Immunohematology Book, John C. Flynn, Jr.
Hemolytic Disease of Newborn (HDN)
ANEMIA

• Once antigen-antibody interaction occurs, antibody-coated red blood
cells are removed from fetal circulation by macrophages of the spleen
and liver which lead to anemia
• Anemia limits the ability of the blood to carry oxygen to the baby's
organs and tissues.

• In response to anemia, fetal bone marrow and other hematopoietic
tissues in the spleen and liver increase the amount of red blood cells
production.
• Baby's responds to the hemolysis by trying to make more red blood
cells very quickly in the bone marrow and the liver and spleen.
ORGAN
• Organs enlarge - hepatosplenomegaly.
ENLARGEME
• New red blood cells released prematurely from bone marrow and are
NT
unable to do the work of mature red blood cells produced many
nucleated red blood cell which is unable to carry oxygen.
• As the red blood cells break down, bilirubin is formed
• Accumulation of bilirubin in body tissue and fluid results in jaundices.
• Due to immature liver of the newborn and inability to conjugated the
HYPERBILIR
unconjugated bilirubin result in the unconjugated bilirubin increasing
UBINEMIA
and crossing the blood brain barrier and cause kernicterus.
KERNICTERUS
• Kernicterus or bilirubin encephalopathy results from high
levels of unconjugated bilirubin in the fetus blood which is
more than 20 mg/dL .
• Because unconjugated bilirubin are lipid soluble and toxic , it
can crosses the blood-brain barrier and it will penetrates
neuronal and glial membrane thus cause neurotoxicity
• Patients surviving kernicterus have severe permanent
neurologic symptoms such as
a) choreoathetosis
b) spasticity
c) muscular rigidity
d) ataxia
e) deafness
f) mental retardation
Kernicterus

webhome.idirect.com/~brainology/braino
logy/devpath_25_kernicterus.html

http://www.nlm.nih.gov/medlineplus/ency/imagepages/1987
5.htm

• Affected structures have a bright yellow color due to bilirubin stained .
HYDROPS FETALIS
As the fetus ‘s anemia worsen, the fetus
accelerates red cells production
 Liver and spleen increase in size due to
increase production of red blood cells.
As a result, portal hypertension occur,
causing the liver to reduces its albumin
production.
 This in turn result in edema.
Edema is an abnormal accumulation of
fluid beneath the skin.
 This condition in the fetus is known as
hydrop fetalis

https://www.capefearvalley.com/outreach/Outreach/Peapods/
HemolyticDiseaseNewborn/RhIncompatibility.htm
CLINICAL SYMPTOM
• Varies from mild jaundice and anemia to hydrops fetalis
(with ascites, pleural and pericardial effusions)
• Chief risk to the fetus is anemia.
• Extramedullary hematopoiesis due to anemia results in
hepatosplenomegaly.
• Risks during labor and delivery include:
• asphyxia and splenic rupture.
• Postnatal problems include:
• Asphyxia
• Pulmonary hypertension
• Pallor (due to anemia)
• Edema (hydrops, due to low serum albumin)
• Respiratory distress
• Coagulopathies (↓ platelets & clotting factors)
• Jaundice
• Kernicterus (from hyperbilirubinemia)
• Hypoglycemia (due to hyperinsulinemnia from islet cell
hyperplasia)
Hemolytic Disease of Newborn (HDN)
Test

Situation

Timing

Pregnancy

Initial visit

ABO Typing
Initial visit & 26-28
weeks
All Pregnancies
Iniital visit
D negative pregancy
Before RhIg therapy
Antibody Screen
3rd trimester if history
D positive pregnancy
of antibodies or
transfusion
Positive antibody screen
Upon detection
Antibody ID
Rh Typing

Pregnancy

Prenatal Testing

Antibody titer

Rh or other clinically
signficant antibody

Upon initial detection
Repeat at 18 – 20
weeks
Repeat at 2-4 week
intervals if below
critical titer
POSTNATAL TESTING
• Determination of maternal ABO and D typing at
delivery.
AMNIOCENTESIS
• Technique use to provide a specimen to assess the severity of
fetal hemolytic disease.
• A needle is introduced through abdomen and uterus and into the
amniotic sacs. A small amount of amniotic fluid is remove for
testing.
• Liley’s graph is use to evaluate the level of bile pigment in the
amniotic fluid to measure hemolysis in the fetus.
• Liley’s graph is used to correct for gestation of less than 27 week
because bilirubin level is normally peak at 23 to 25 week of
gestation in unaffected fetus.
Liley’s Graph

Liley’s bottom zone represent affected or very mildy
affected fetus. The middle zone show minimally
affected fetus and the upper zone indicate moderate
to marked hemolysis in fetus. Higher concentration
of bilirubin in amniotic fluid is found in abnormal fetus.
Hemolytic Disease of Newborn (HDN)
Management of
HDN

Prevention

Treatment

Maternal

Plasma
Exchange

Intravenous
Immune
Globulin

Infant

Intrauterine
transfusion

Rh Immune
Globulin (RhIG or
Rhogam)

Exchage
transfusion

Phototherapy
TREATMENT
Intrauterine Transfusion (IUT)
• Red Blood Cells (RBCs) are infused into abdominal cavity of fetus and then absorbed
into fetal circulation to avoid hydrops fetalis and fetal death.
• Can be done as early as 17 weeks.
• In intraperitoneal IUT, a needle is passed through the mother’s abdomen and into the
abdomen of the fetus by the help of ultrasound image to determine the position of the
fetus and placenta.
• Selection of Blood for IUT:
- Most IUTs are accomplished using group O.
- Rh-negative RBCs that are less then 7 days from collection
- Ctyomegalovirus (CMV) antibody negative
- Hemoglobin S negative
- Fresh blood is used to provide RBCs with longest viability
and to avoid lower pH, decreased 2,3-diphospoglycerate, and
elevated potassium levels.
- RBCs are usually dry packed to remove residual anti-A and anti-B and reconstituted
with group AB fresh frozen plasma t provide coagulation factors.
Intrauterine Transfusion (IUT)
• The risk of these procedures is now largely dependent on the prior condition of the
fetus and the gestational age at which transfusion is commenced.
• Titer greater than 32 for anti-D and 8 for anti-K OR four fold increase in titer
indicates need for analysis of amniotic fluid.
• Amniocentesis
• Perform at 28 wks if HDN in previous child
• Perform at 22 wks if previous child severely affected
• Perform if maternal antibody increases before 34th wk.
• High values of bilirubin in amniotic fluid analyses by the Liley method or a
hemoglobin concentration of cord blood below 10.0 g/mL.
Liley Chart
IUT Procedure
Postpartum - Phototherapy
• Phototherapy is the treatment of choice.
• Phototherapy accelerates bilirubin metabolism through the
process of photodegradation .
• The effective wavelength of light seeems to be in te 420- to 475
nm range.
• During photoisomerization or photooxidation, the insoluble form of
unconjugated bilirubin is converted into water soluble form which
permits more rapid excretion without conjugation, through the bile
or urine.
Phototherapy
• However phototherapy may not be effective in
controlling hyperbillirubinemia secondry to HDN if
the concentration of bilirubin rises at rate of 0.5 to
2.0 mg/dl per hour. In the cases, exchange
transfusion must occur.
Exchange transfusion
Objective:
1.
2.
3.
4.

To decrease the level of bilirubin and prevent kernicterus.
To remove baby’s sensitized red blood cells.
To provide compatible red blood cells adequate oxygen carrying
capacity.
To decrease the level of incompatible antibody in the baby.
• Full-term infants rarely require an exchange transfusion if
the infant undergo the phototherapy.
• The exchange transfusion is done if the total bilirubin level
is aprroaching 20mg/dL and continues to rise despite the
baby undergo the phototherapy.
• The blood should be reconstituted from fresh, O negative
packed RBCs cross-matched against the mother and typespecific fresh frozen plasma.

Continue….
Technique uses:
• 30 min before the exchange transfusion, give albumin 1g/kg
to increase the bilirubin bound to albumin in the circulation
and make the exchange transfusion more effective.
• Exchange 2 times the blood volume, at 85mL/kg, by using
the isovolumic technique. This technique done by
withdrawing blood from upper atrium canal and infusing
through upper ventricle canal(low right atrium with tip intra
ventricle canal).
• Do not infuse blood through upper ventricle canal if tips is in
portal circulation.
• The blood should be warmed and the bag agitated every few
minutes to prevent settling of the RBCs.
Plasma Exchange
• It is widely used treatment of immune-mediated disease.
• This treatment is applied to the pregnant women with
high antibody titer, or that has past history of stillbirth due
to HDN.
• This procedure are effective in decreasing the antibody
titer and quantity of antibody.
• Plasma exchange can reduce antibody titer up to 75%.
• Beside that, it also use for a way to delay the need for
fetal intervention that has been hydrops fetalis (edema) in
which before 22 week gestation in a previous pregnancy.
Intravenous Immune Globulin
• Intravenous immune globulin (IVIG) is made up from
plasma isolated
• This treatment use strengthen body immune system
beside to treat immune deficiency
• Intravenous Immunoglobulins were found to
decrease hemolysis leading to reduction in serum
bilirubin level.
• The immunoglobulin could act by occupying the FC
receptors of reticulo-endothelial cells preventing
them from taking up and lysing antibody coated
RBCs. This subsequently leads to decrease in the
need for exchange transfusion.
PREVENTION
Rh Immune Globulin (RhIG or Rhogam)
• Alloimmunization in pregnant woman can be prevented by
administration of Rh immune globulin (RhIG or Rhogam).
• RhIG is a concentrate of IgG anti-D prepared from pooled
human plasma of D-negative people who have been exposed to
the D antigen and who have made antibodies to it.
• Significance of administration of RhIG
• (i) RhIG prevents alloimmunization in D-negative mother
exposed to D-positive fetal red cells. In the meantime, it protects
mother from being sensitized to D antigen of fetal during
pregnancy and after delivery of infant.
• (ii) RhIG suppresses mother’s immune response following
exposure to D-positive fetal red cells.
• (iii) RhIG prevents the mother from producing anti-D.
• (iv) RhIG protects subsequent D-positive pregnancies but must
be given each time the woman is pregnant with D-positive fetus.
• Principle
 RhIG is given by injection into mother’s muscle
(intramuscular).
 Giving RhIG to D-negative pregnant woman prevents her
immune system from producing its own anti-D (which would
attack her D-positive fetal red cells).
 When RhIG is introduced into D-negative mothers’s
bloodstream, RhIG antibodies will locate D-positive fetal red
cells.
 RhIG antibodies will attach to D-positive fetal red cells and
covering their presence from mother’s immune system.
 Therefore, sensitization of anti-D to D-positive fetal red cells
can be prevented.
Retrieved from http://www.carolguze.com/text/102-13-humangenetics.shtml
• Guidelines for RhIG administration in D-negative pregnant woman :
(i) All doses should be given within 72 hours of delivery or procedure. If she is
not given RhIG within 72 hours after the birth of an Rh positive baby, she will
begin to make antibodies to the fetal blood cells.
(ii) Woman should not be sensitized to D antigen.
iii) 50 µg dose - Up to 12th week of gestation for abortion, miscarriage and end
period of ectopic pregnancy.
(iv) 120 µg dose
- End period of pregnancy which is after 12th week of gestation.
- After amniocentesis.
- After delivery of D-positive infant.
(v) 300 µg dose
- At 28th week of gestation
- After amniocentesis
- After delivery of D-positive infant.
REFERENCES
• Quinley, E. D. (1998). Hemolytic Disease of Newborn. In
E. D. Quinley, Immunohematology Principle and
Practice (pp. 282-283). Philadelphia: Lippincott-Raven.
• Quinley, E. D. (1998). Hemolytic Disease of
Newborn. In E. D. Quinley, Immunohematology
Principle and Practice
(pp. 282-283). Philadelphia:
Lippincott- Raven.

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Hemolytic Disease of Newborn (HDN)

  • 1. MLT 502 – IMMUNOHAEMATOLOGY II HS221/5 GROUP C HEMOLYTIC DISEASE OF NEWBORN (HDN)
  • 2. DEFINITION A condition in which fetus or neonate’s red blood cell (RBC) are destroyed by Immunoglobulin G (IgG) antibodies produced by mother.  "hemolytic" means breaking down of red blood cells  "erythroblastosis" refers to making of immature red blood cells  "fetalis" refers to fetus
  • 3. HISTORY Haemolytic disease of the newborn (HDN) has been known for centuries with its typical and poor prognosis. The first described by a French midwife who delivered twins in 1609. One baby was swollen and died soon after birth, the other baby developed jaundice and died several days later. For the next 300 years, many similar cases were described in which newborns failed to survive. During 1970s, the routine antenatal care included screening of all expectant mothers to find those whose pregnancy may be at risk of HDN, and giving preventative treatment accordingly. This has led to decrease cases in the incidence of HDN, particularly severe cases that were responsible for stillbirth and neonatal death.
  • 5. CAUSES OF HDN Rh INCOMPATIBILITY ABO INCOMPATIBILITY OTHER SYSTEM ANTIBODIES INCOMPATIBILITY OTHER Rh ANTIBODIES OTHER ANTIBODIES
  • 6. CAUSES OF HDN 1) Rh incompatibility - HDN is occured when a mother with Rh-negative blood becomes pregnant with Rh-positive baby that inherited from Rh-positive father. - It occurs when anti-D is stimulated in mother plasma due to mother ‘s immune response to the antigen D on fetal’s red blood cells. - This is due to anti-D is an IgG that capable to cross placenta and hence delivered to fetal circulation. - Rh caused HDN is less common but more severe. 2) ABO incompatibility - HDN is arouse when a mother with blood type O becomes pregnant with a fetus with different blood types A, B or AB. - ABO antibodies is natural occurring antibodies that clinically significant. - ABO caused HDN is commonly occur but less severe.
  • 7. 3) Others unexpected immune antibody (other than anti-D) : - Other Rh antibodies : - anti-E (second most common, mild disease) - anti–c (third most common, mild to severe) - anti-C and anti-e (rare) - antibody combination (anti-c and anti-E occurring together, can be severe) - Other antibodies - Kell system antibodies (uncommon causes) - Duffy, MNSs and Kidd system antibodies (rare causes) * Lewis and P system antibody are no occurrence in HDN.
  • 9. PHYSIOLOGIC OF HDN Physiologic of HDN is defined as process of how HDN occur (mechanism).
  • 10. RHESUS HEMOLYTIC DISEASE OF NEWBORN • In a first pregnancy, Rh sensitization is not likely. Usually it only becomes a problem in a future pregnancy with another Rh positive baby. • When the next pregnancy occur, the mother's antibodies cross the placenta then reacts with an RBC antigen to fight the Rh positive cells in the baby's body that the baby has inherited from the father, and that is foreign to the mother. • Hence, the antigen antibody interaction occurs.
  • 11. • Sensitization of baby’s red blood cell (RBC) by mother’s IgG antibody causes the baby’s RBC to be destroyed. • These antibody-coated RBCs are removed from fetal circulation by the macrophages of the spleen and liver. • The severity of anemia depends on the amount of mother’s antibody, its specificity, its avidity, and others characteristics. • Anemia will stimulate bone marrow to produce more RBC including immature RBC, which is then released to fetus circulation. This is also known as erythroblastosis fetalis.
  • 12. This figure shows pathophysiology of HDN Retrieved from: http://tmedweb.tulane.edu/pharmwiki/lib/exe/fetch.php/rhod.png
  • 13. This figure explains the Mechanism of HDN Retrieved from: http://legacy.owensboro.kctcs.edu/gcaplan/anat2/notes/14_23.jpg
  • 14. ABO INCOMPATIBILITY OF HDN • ABO HDN is due to incompatibility of ABO between mother and fetus. • A portion of the mother’s ABO antibodies may be IgG and cross the placenta. • It thus sensitizes the fetal cells. • This condition can occur on the first pregnancy and can occur without there having been transfusion. • The mechanism of ABO HDN is similar to the RH HDN. • Occurrence of ABO HDN is somewhat common, but fortunately it is not often very severe.
  • 15. This figure shows circulating IgG maternal antibodies cross the placenta and sensitize fetal antigens positive cells. Retrieved from Essential of Immunohematology Book, John C. Flynn, Jr.
  • 17. ANEMIA • Once antigen-antibody interaction occurs, antibody-coated red blood cells are removed from fetal circulation by macrophages of the spleen and liver which lead to anemia • Anemia limits the ability of the blood to carry oxygen to the baby's organs and tissues. • In response to anemia, fetal bone marrow and other hematopoietic tissues in the spleen and liver increase the amount of red blood cells production. • Baby's responds to the hemolysis by trying to make more red blood cells very quickly in the bone marrow and the liver and spleen. ORGAN • Organs enlarge - hepatosplenomegaly. ENLARGEME • New red blood cells released prematurely from bone marrow and are NT unable to do the work of mature red blood cells produced many nucleated red blood cell which is unable to carry oxygen. • As the red blood cells break down, bilirubin is formed • Accumulation of bilirubin in body tissue and fluid results in jaundices. • Due to immature liver of the newborn and inability to conjugated the HYPERBILIR unconjugated bilirubin result in the unconjugated bilirubin increasing UBINEMIA and crossing the blood brain barrier and cause kernicterus.
  • 18. KERNICTERUS • Kernicterus or bilirubin encephalopathy results from high levels of unconjugated bilirubin in the fetus blood which is more than 20 mg/dL . • Because unconjugated bilirubin are lipid soluble and toxic , it can crosses the blood-brain barrier and it will penetrates neuronal and glial membrane thus cause neurotoxicity • Patients surviving kernicterus have severe permanent neurologic symptoms such as a) choreoathetosis b) spasticity c) muscular rigidity d) ataxia e) deafness f) mental retardation
  • 20. HYDROPS FETALIS As the fetus ‘s anemia worsen, the fetus accelerates red cells production  Liver and spleen increase in size due to increase production of red blood cells. As a result, portal hypertension occur, causing the liver to reduces its albumin production.  This in turn result in edema. Edema is an abnormal accumulation of fluid beneath the skin.  This condition in the fetus is known as hydrop fetalis https://www.capefearvalley.com/outreach/Outreach/Peapods/ HemolyticDiseaseNewborn/RhIncompatibility.htm
  • 21. CLINICAL SYMPTOM • Varies from mild jaundice and anemia to hydrops fetalis (with ascites, pleural and pericardial effusions) • Chief risk to the fetus is anemia. • Extramedullary hematopoiesis due to anemia results in hepatosplenomegaly. • Risks during labor and delivery include: • asphyxia and splenic rupture. • Postnatal problems include: • Asphyxia • Pulmonary hypertension • Pallor (due to anemia) • Edema (hydrops, due to low serum albumin) • Respiratory distress • Coagulopathies (↓ platelets & clotting factors) • Jaundice • Kernicterus (from hyperbilirubinemia) • Hypoglycemia (due to hyperinsulinemnia from islet cell hyperplasia)
  • 23. Test Situation Timing Pregnancy Initial visit ABO Typing Initial visit & 26-28 weeks All Pregnancies Iniital visit D negative pregancy Before RhIg therapy Antibody Screen 3rd trimester if history D positive pregnancy of antibodies or transfusion Positive antibody screen Upon detection Antibody ID Rh Typing Pregnancy Prenatal Testing Antibody titer Rh or other clinically signficant antibody Upon initial detection Repeat at 18 – 20 weeks Repeat at 2-4 week intervals if below critical titer
  • 24. POSTNATAL TESTING • Determination of maternal ABO and D typing at delivery.
  • 25. AMNIOCENTESIS • Technique use to provide a specimen to assess the severity of fetal hemolytic disease. • A needle is introduced through abdomen and uterus and into the amniotic sacs. A small amount of amniotic fluid is remove for testing. • Liley’s graph is use to evaluate the level of bile pigment in the amniotic fluid to measure hemolysis in the fetus. • Liley’s graph is used to correct for gestation of less than 27 week because bilirubin level is normally peak at 23 to 25 week of gestation in unaffected fetus.
  • 26. Liley’s Graph Liley’s bottom zone represent affected or very mildy affected fetus. The middle zone show minimally affected fetus and the upper zone indicate moderate to marked hemolysis in fetus. Higher concentration of bilirubin in amniotic fluid is found in abnormal fetus.
  • 29. TREATMENT Intrauterine Transfusion (IUT) • Red Blood Cells (RBCs) are infused into abdominal cavity of fetus and then absorbed into fetal circulation to avoid hydrops fetalis and fetal death. • Can be done as early as 17 weeks. • In intraperitoneal IUT, a needle is passed through the mother’s abdomen and into the abdomen of the fetus by the help of ultrasound image to determine the position of the fetus and placenta. • Selection of Blood for IUT: - Most IUTs are accomplished using group O. - Rh-negative RBCs that are less then 7 days from collection - Ctyomegalovirus (CMV) antibody negative - Hemoglobin S negative - Fresh blood is used to provide RBCs with longest viability and to avoid lower pH, decreased 2,3-diphospoglycerate, and elevated potassium levels. - RBCs are usually dry packed to remove residual anti-A and anti-B and reconstituted with group AB fresh frozen plasma t provide coagulation factors.
  • 30. Intrauterine Transfusion (IUT) • The risk of these procedures is now largely dependent on the prior condition of the fetus and the gestational age at which transfusion is commenced. • Titer greater than 32 for anti-D and 8 for anti-K OR four fold increase in titer indicates need for analysis of amniotic fluid. • Amniocentesis • Perform at 28 wks if HDN in previous child • Perform at 22 wks if previous child severely affected • Perform if maternal antibody increases before 34th wk. • High values of bilirubin in amniotic fluid analyses by the Liley method or a hemoglobin concentration of cord blood below 10.0 g/mL. Liley Chart IUT Procedure
  • 31. Postpartum - Phototherapy • Phototherapy is the treatment of choice. • Phototherapy accelerates bilirubin metabolism through the process of photodegradation . • The effective wavelength of light seeems to be in te 420- to 475 nm range. • During photoisomerization or photooxidation, the insoluble form of unconjugated bilirubin is converted into water soluble form which permits more rapid excretion without conjugation, through the bile or urine.
  • 32. Phototherapy • However phototherapy may not be effective in controlling hyperbillirubinemia secondry to HDN if the concentration of bilirubin rises at rate of 0.5 to 2.0 mg/dl per hour. In the cases, exchange transfusion must occur.
  • 33. Exchange transfusion Objective: 1. 2. 3. 4. To decrease the level of bilirubin and prevent kernicterus. To remove baby’s sensitized red blood cells. To provide compatible red blood cells adequate oxygen carrying capacity. To decrease the level of incompatible antibody in the baby.
  • 34. • Full-term infants rarely require an exchange transfusion if the infant undergo the phototherapy. • The exchange transfusion is done if the total bilirubin level is aprroaching 20mg/dL and continues to rise despite the baby undergo the phototherapy. • The blood should be reconstituted from fresh, O negative packed RBCs cross-matched against the mother and typespecific fresh frozen plasma. Continue….
  • 35. Technique uses: • 30 min before the exchange transfusion, give albumin 1g/kg to increase the bilirubin bound to albumin in the circulation and make the exchange transfusion more effective. • Exchange 2 times the blood volume, at 85mL/kg, by using the isovolumic technique. This technique done by withdrawing blood from upper atrium canal and infusing through upper ventricle canal(low right atrium with tip intra ventricle canal). • Do not infuse blood through upper ventricle canal if tips is in portal circulation. • The blood should be warmed and the bag agitated every few minutes to prevent settling of the RBCs.
  • 36. Plasma Exchange • It is widely used treatment of immune-mediated disease. • This treatment is applied to the pregnant women with high antibody titer, or that has past history of stillbirth due to HDN. • This procedure are effective in decreasing the antibody titer and quantity of antibody. • Plasma exchange can reduce antibody titer up to 75%. • Beside that, it also use for a way to delay the need for fetal intervention that has been hydrops fetalis (edema) in which before 22 week gestation in a previous pregnancy.
  • 37. Intravenous Immune Globulin • Intravenous immune globulin (IVIG) is made up from plasma isolated • This treatment use strengthen body immune system beside to treat immune deficiency • Intravenous Immunoglobulins were found to decrease hemolysis leading to reduction in serum bilirubin level. • The immunoglobulin could act by occupying the FC receptors of reticulo-endothelial cells preventing them from taking up and lysing antibody coated RBCs. This subsequently leads to decrease in the need for exchange transfusion.
  • 38. PREVENTION Rh Immune Globulin (RhIG or Rhogam) • Alloimmunization in pregnant woman can be prevented by administration of Rh immune globulin (RhIG or Rhogam). • RhIG is a concentrate of IgG anti-D prepared from pooled human plasma of D-negative people who have been exposed to the D antigen and who have made antibodies to it. • Significance of administration of RhIG • (i) RhIG prevents alloimmunization in D-negative mother exposed to D-positive fetal red cells. In the meantime, it protects mother from being sensitized to D antigen of fetal during pregnancy and after delivery of infant. • (ii) RhIG suppresses mother’s immune response following exposure to D-positive fetal red cells. • (iii) RhIG prevents the mother from producing anti-D. • (iv) RhIG protects subsequent D-positive pregnancies but must be given each time the woman is pregnant with D-positive fetus.
  • 39. • Principle  RhIG is given by injection into mother’s muscle (intramuscular).  Giving RhIG to D-negative pregnant woman prevents her immune system from producing its own anti-D (which would attack her D-positive fetal red cells).  When RhIG is introduced into D-negative mothers’s bloodstream, RhIG antibodies will locate D-positive fetal red cells.  RhIG antibodies will attach to D-positive fetal red cells and covering their presence from mother’s immune system.  Therefore, sensitization of anti-D to D-positive fetal red cells can be prevented.
  • 41. • Guidelines for RhIG administration in D-negative pregnant woman : (i) All doses should be given within 72 hours of delivery or procedure. If she is not given RhIG within 72 hours after the birth of an Rh positive baby, she will begin to make antibodies to the fetal blood cells. (ii) Woman should not be sensitized to D antigen. iii) 50 µg dose - Up to 12th week of gestation for abortion, miscarriage and end period of ectopic pregnancy. (iv) 120 µg dose - End period of pregnancy which is after 12th week of gestation. - After amniocentesis. - After delivery of D-positive infant. (v) 300 µg dose - At 28th week of gestation - After amniocentesis - After delivery of D-positive infant.
  • 42. REFERENCES • Quinley, E. D. (1998). Hemolytic Disease of Newborn. In E. D. Quinley, Immunohematology Principle and Practice (pp. 282-283). Philadelphia: Lippincott-Raven. • Quinley, E. D. (1998). Hemolytic Disease of Newborn. In E. D. Quinley, Immunohematology Principle and Practice (pp. 282-283). Philadelphia: Lippincott- Raven.