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EB VIRUSES
Epstein-Barr Virus (EBV)
• Spread through oral transmission
(SALIVA)
• Infectious Mononucleosis.
• Other Diseases include:
– African or Burkitt’s lymphoma
– Nasopharyngeal carcinoma
– B cell lymphoma
Viral Antigens
• There are three classes :1.Latent phase(EBNAs & LMP)
2.Early phase(proteins)
3.late phase
Epstein-Barr virus
• African or Burkitt’s Lymphoma
– malignant B-cell neoplasm
– presents as a rapidly growing tumour of
the jaw, face or eye
– grows very quickly, and without treatment
most children die within a few months
– Epstein-Barr virus (EBV) has been
strongly implicated
African or Burkitt’s Lymphoma
• Although BL is a very rapidly growing tumour it
responds well to treatment.
• Three pictures: before treatment, 3 days and 6
days after treatment
Nasopharyngeal Carcinoma
• Endemic in South China, Africa, Arctic
Eskimos
• This is a malignant tumour of the squamous
epithelium of the nasopharynx.
• 100% contain EBV DNA
• Rates are less than 1 per 100,000 in most
populations
• Nasopharyngeal carcinomas are found in
association with reactivation of latent
Epstein-Barr Virus.
• The exact mechanisms of association are
unknown
B-Cell Lymphoma
• In most individuals infected with EBV, the
virus is present in the B-cells, which are
normally controlled by T-lymphocytes
• When T-cell deficiency exists, one clone of
EBV-infected B-lymphocytes escapes
immune surveillance to become
autonomously proliferating.
• EBV induced B cell lymphomas are most
prevalent in immunocompromised patients.
Oral Hairy Cell Leukoplakia
• Viral infection of the oral cavity.
• Indicator of HIV infection as well as of a
person's lessening or weakening immunity
Infectious Mononucleosis
•
•
•
•
•
•

4 to 7 week incubation
Acute self-limiting infection of the RE system
Enlarged lymph nodes in the neck.
Sore throat, fever, rash
Malaise, lethargy, extreme tiredness
Liver and spleen involvement and
enlargement
• Hematology: High WBC, over 20% atypical
reactive lymphocytes also known as Downey
cells.
Infectious Mononucleosis
• Downey cells may be present
Heterophile Antigens/Antibodies
• Heterophile antigens are a group of
similar antigens found in unrelated
animals, IE, man, sheep, horse, dog
cat, mouse.
• Heterophile antibodies produced
against heterophile antigens of one
species will cross react with others.
Heterophile Antigens/Antibodies
• Forssman antigen is an example of a
heterophile antigen and is found on the
RBCs of many species (guinea pig,
dog, cat, mouse, sheep, fowl, horse)
• Forssman antibodies formed against
Forssman antigens will agglutinate
sheep RBCs.
Paul Bunnell Test
• The original Paul-Bunnell test was a simple titration of sheep
cell agglutinins but this procedure was subsequently modified
in order to distinguish between sheep cell agglutinins formed in
IM and the Forssman-type antibodies found in normal serum,
serum sickness and in certain other conditions.
• Tissues rich in Forssman antigen (guinea pig kidney) absorb
Forssman antibodies but do not affect the heterophil antibodies
in IM.
• Heterophil antibodies are absorbed by beef cells,
• Forssman hapten is a glycolipid usually associated with a
protein, the determinant being largely carbohydrate and
therefore heat stable.
Davidsohn Differential
• The principle behind the Paul-Bunnell-Davidsohn test is that
the two types of sheep agglutinins are distinguished by titrating
them before and after absorption with guinea pig kidney and ox
cells.
• Patients serum containing antibodies due to IM is added to
guinea pig kidney cells. These antibodies are not absorbed by
the kidney cells. These antibodies then react with Beef (Ox)
red blood cells which causes agglutination and is a positive test
for IM.
• Patients serum containing Forssman antibodies are added to
guinea pig kidney cells. Antibodies are absorbed by the kidney
cells. These antibodies are then allowed to react with Beef red
blood cells which does not cause agglutination. This is a
positive test for Forssman antigens.
Davidsohn Differential
* To be considered absorbed there must be greater than a three tube difference
between the presumptive titer and the differential titer.

Heterophil Antibody
------------------------

Kidney Extract
------------------

Beef Erythrocyte
---------------------

Infectious Mono

Not Absorbed

Absorbed

Forssman

Absorbed

Not Absorbed

Serum Sickness

Absorbed

Absorbed
Davidsohn Differential
Advantages
•

When properly performed, this
test is specific for Infectious
Mononucleosis and falsepositive results are rare.

Disadvantages
•

Davidsohn Differential test is
very time consuming and
burdensome.
Infectious Mono Slide Tests
• It was discovered that horse RBCs
possess antigens which react with the
antibody associated with IM.
• Patient serum mixed with horse RBCs,
agglutination is positive.
• Latex agglutination
• Not diagnostic, must look at total
clinical picture.
EBV Specific Antibodies
• EBV specific antibodies may be measured.
• Pattern of appearance of EBV antigens.
• Most valuable is IgM antibody to viral capsid
antigen (VCA), indicates a current infection
(best marker), lasts about 12 weeks.
• Can also detect anti-early antigen (EA)
(recent infection) and anti EB nuclear antigen
(EBNA) (older infection).
• ELISA and IFA most commonly used

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epstein-bar virus (EBV)

  • 2. Epstein-Barr Virus (EBV) • Spread through oral transmission (SALIVA) • Infectious Mononucleosis. • Other Diseases include: – African or Burkitt’s lymphoma – Nasopharyngeal carcinoma – B cell lymphoma
  • 3. Viral Antigens • There are three classes :1.Latent phase(EBNAs & LMP) 2.Early phase(proteins) 3.late phase
  • 4. Epstein-Barr virus • African or Burkitt’s Lymphoma – malignant B-cell neoplasm – presents as a rapidly growing tumour of the jaw, face or eye – grows very quickly, and without treatment most children die within a few months – Epstein-Barr virus (EBV) has been strongly implicated
  • 5. African or Burkitt’s Lymphoma • Although BL is a very rapidly growing tumour it responds well to treatment. • Three pictures: before treatment, 3 days and 6 days after treatment
  • 6. Nasopharyngeal Carcinoma • Endemic in South China, Africa, Arctic Eskimos • This is a malignant tumour of the squamous epithelium of the nasopharynx. • 100% contain EBV DNA • Rates are less than 1 per 100,000 in most populations • Nasopharyngeal carcinomas are found in association with reactivation of latent Epstein-Barr Virus. • The exact mechanisms of association are unknown
  • 7. B-Cell Lymphoma • In most individuals infected with EBV, the virus is present in the B-cells, which are normally controlled by T-lymphocytes • When T-cell deficiency exists, one clone of EBV-infected B-lymphocytes escapes immune surveillance to become autonomously proliferating. • EBV induced B cell lymphomas are most prevalent in immunocompromised patients.
  • 8. Oral Hairy Cell Leukoplakia • Viral infection of the oral cavity. • Indicator of HIV infection as well as of a person's lessening or weakening immunity
  • 9. Infectious Mononucleosis • • • • • • 4 to 7 week incubation Acute self-limiting infection of the RE system Enlarged lymph nodes in the neck. Sore throat, fever, rash Malaise, lethargy, extreme tiredness Liver and spleen involvement and enlargement • Hematology: High WBC, over 20% atypical reactive lymphocytes also known as Downey cells.
  • 10. Infectious Mononucleosis • Downey cells may be present
  • 11. Heterophile Antigens/Antibodies • Heterophile antigens are a group of similar antigens found in unrelated animals, IE, man, sheep, horse, dog cat, mouse. • Heterophile antibodies produced against heterophile antigens of one species will cross react with others.
  • 12. Heterophile Antigens/Antibodies • Forssman antigen is an example of a heterophile antigen and is found on the RBCs of many species (guinea pig, dog, cat, mouse, sheep, fowl, horse) • Forssman antibodies formed against Forssman antigens will agglutinate sheep RBCs.
  • 13. Paul Bunnell Test • The original Paul-Bunnell test was a simple titration of sheep cell agglutinins but this procedure was subsequently modified in order to distinguish between sheep cell agglutinins formed in IM and the Forssman-type antibodies found in normal serum, serum sickness and in certain other conditions. • Tissues rich in Forssman antigen (guinea pig kidney) absorb Forssman antibodies but do not affect the heterophil antibodies in IM. • Heterophil antibodies are absorbed by beef cells, • Forssman hapten is a glycolipid usually associated with a protein, the determinant being largely carbohydrate and therefore heat stable.
  • 14. Davidsohn Differential • The principle behind the Paul-Bunnell-Davidsohn test is that the two types of sheep agglutinins are distinguished by titrating them before and after absorption with guinea pig kidney and ox cells. • Patients serum containing antibodies due to IM is added to guinea pig kidney cells. These antibodies are not absorbed by the kidney cells. These antibodies then react with Beef (Ox) red blood cells which causes agglutination and is a positive test for IM. • Patients serum containing Forssman antibodies are added to guinea pig kidney cells. Antibodies are absorbed by the kidney cells. These antibodies are then allowed to react with Beef red blood cells which does not cause agglutination. This is a positive test for Forssman antigens.
  • 15. Davidsohn Differential * To be considered absorbed there must be greater than a three tube difference between the presumptive titer and the differential titer. Heterophil Antibody ------------------------ Kidney Extract ------------------ Beef Erythrocyte --------------------- Infectious Mono Not Absorbed Absorbed Forssman Absorbed Not Absorbed Serum Sickness Absorbed Absorbed
  • 16. Davidsohn Differential Advantages • When properly performed, this test is specific for Infectious Mononucleosis and falsepositive results are rare. Disadvantages • Davidsohn Differential test is very time consuming and burdensome.
  • 17. Infectious Mono Slide Tests • It was discovered that horse RBCs possess antigens which react with the antibody associated with IM. • Patient serum mixed with horse RBCs, agglutination is positive. • Latex agglutination • Not diagnostic, must look at total clinical picture.
  • 18. EBV Specific Antibodies • EBV specific antibodies may be measured. • Pattern of appearance of EBV antigens. • Most valuable is IgM antibody to viral capsid antigen (VCA), indicates a current infection (best marker), lasts about 12 weeks. • Can also detect anti-early antigen (EA) (recent infection) and anti EB nuclear antigen (EBNA) (older infection). • ELISA and IFA most commonly used