ANTINUCLEAR ANTIBODY

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ANTINUCLEAR ANTIBODY

  1. 1. IRUM RAFIQ SHAIKHANTINUCLEAR ANTIBODY
  2. 2. ANTINUCLEAR ANTIBODYAnti-nuclear antibodies (ANAs, also known asanti-nuclear factor or ANF) are autoantibodiesthat bind to contents of the cell nucleus. Innormal individuals, the immune system producesantibodies to foreign proteins (antigens) but notto human proteins (autoantigens). In someindividuals, antibodies to human antigens areproduced
  3. 3. Subtypes Of ANAsThere are many subtypes of ANAs such as, anti-Ro antibodies anti-La antibodies anti-Sm antibodies anti-nRNP antibodies anti-Scl-70 antibodies anti-dsDNA antibodiesanti-histone antibodies antibodies to nuclear pore complexes anti-centromere antibodies anti-sp100 antibodies
  4. 4. Subtypes Of ANAsEach of these antibody subtypes binds todifferent proteins or protein complexeswithin the nucleus. They are found in manydisorders including autoimmunity, cancerand infection, with different prevalence ofantibodies depending on the condition.
  5. 5. Nuclear ProteinsProteins that have beenSynthesized in the nucleusAnd thereafter whereDistributed to theirrespective sites in the cell
  6. 6. Detection TechniqueIndirect immunofluorescence is the referencemethod for screening and titration ofcirculating autoantibodies in human serum.Using three different tissues fromrat(liver,kidney and stomach) enablesautontibodies to be more easily identifying bycomparing the results obtained with each tissue.
  7. 7. Types of ANAsThere are Three major antibodiesdetected Antinuclear antibodies (ANA)Antimitochondrial antibodies (AMA)Antismooth muscle antibodies(ASMA)
  8. 8. PrincipleAn indirect immunofluorescence technique isutilized where patient samples and appropriatecontrols are incubated with the substrate slides.The unreacted antibodies are washed off and anappropriate fluorescence labeled conjugate isapplied.Unbound conjugate is washed off, and slides areviewed with a fluorescence microscope.Positive samples produce apple-greenfluorescence.
  9. 9. PATTERN OF FLUORESCENCESpeckledHomogenousRimNucleolar
  10. 10. PATTERN COMMON ANTIGENINVOLVEDDISEASE ASSOCIATIONHomogenous dsDNA,histones SLE,RAMixed connective tissuedisease,Drug induced lupusPeripheral (RIM) Native/dsDNA SLESpeckled Extractable nuclear antigens,Ribonucleoprotein,Scl-70,SSBSclerodermaSjogren’s syndrome, Mixedconnective tissue disease,SLENucleolar 4-6S sRNA Scleroderma,SLE,RASjogren’s syndrome,Progressive systemic sclerosis.
  11. 11. Importance Of ANASerologic hallmarks of patients withsystemic autoimmune disease (ANAdiseases).Can provide further diagnostic andprognostic data concerning patients whohave minimal symptoms or who haveclinical features of more than oneautoimmune disease.
  12. 12. Limitations of ANATheir presence does not mandate the presence ofillness, since they can also be found in otherwisenormal individuals.Accurate interpretation of different nuclearpatterns is confounded several difficulties as:One nuclear pattern may obscure and preventthe recognition of another pattern if severalantibodies are present simultaneously.

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