Laboratory examinations in autoimmune disorders

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Laboratory examinations in autoimmune disorders

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• ??????????

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• SLE ANA, Anti-dsDNA, Anti-Sm
• Drug-induced lupus Anti-histone
• Rheumatoid arthritis RF, ACPA (Anti- CCP)
• Mixed connective tissue disease (MCTD) Anti-RNP
• Polymyositis Anti-Jo-1

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• Scleroderma Anti-Scl-70, Anti-centromere
• Wegeners granulomatosis c-ANCA
• Microscopic polyangiitis p-ANCA
• Sjogrens syndrome Anti-Ro (SSA), Anti-La (SSB) 
  
• Antiphospholipid syndrome Anti-cardiolipin,
  lupus anticoagulant, Anti-beta2-glycoprotein-I Ab

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Frequency of antibodies in various autoimmune
diseases
ANA Anti-ds DNA Anti-Sm Anti-RNP Anti- SSA-Ro Anti- SSB-La Anti-histone Anti- centromere Anti- Jo-1 Anti- Scl-70
SLE gt95? 50-80? 25-40? 26-45? 30-40? 0-15? 25-60? - -   -  
Drug-induced lupus gt95? Rare Rare - - - 90? - -   -  
Mixed connective tissue disease 99? Rare Rare 100? Rare 0-20? - - - -
Rheumatoid arthritis 20-50? Rare Rare 10? 5-7? - 20? - -   -  
Sjogren's syndrome 20-50? - Rare Rare 20-70? 33-60? - - -   -  
Scleroderma 30-50? Rare Rare 22? 0-7? Rare - (CREST) 89-90? - 70
Polymyositis 20-30? Rare Rare Rare 0-9? Rare - - 25-50? -
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Antinuclear antibodies (ANAs)

• ANA?????????????????????
• ??
• nucleic acids (DNA, RNA)
• nucleoproteins found in the connective tissue
  diseases

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Antinuclear antibodies (ANAs)

• ???ANA??1 40 or 1 80,?????????????????
• ???????(gt640?),??SLE?scleroderma?MCTD????????????
  ,???????????,????????????????
• ???????????????????

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Antinuclear antibodies (ANAs)

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• ???????????????,?????????,????????????,?????
  ??,???????180,1160,1320..?????????,????????????
  ?????????????Hep-2????????????,???????Hep-2???????
  ????

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Antinuclear antibodies (ANAs)

• Homogeneous (diffuse)
• 1. Anti-ds DNA, Histones, Nucleosomes, Ku
• -- SLE, drug-induced SLE

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Antinuclear antibodies (ANAs)

• Speckled pattern
• 1. Anti-RNP, Anti-Sm (ribonucleoprotein, a
  component of splicesome)
• -- MCTD, SLE
• 2. SSA (Ro)/ SSB (La) (transcription
  termination factor)
• -- Sjögrens syndrome
• -- Subacute cutaneous lupus

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Antinuclear antibodies (ANAs)

• Nucleolar patterns
• 1. Homogenous nucleolar pattern- anti-PM/ Scl
• --Systemic sclerosis, Polymyositis or
  Dermatomyositis
• 2. Clumpy nucleolar pattern- anti-fibrillarin
• -- Systemic sclerosis
• 3. Fine punctate nucleolar pattern-
  Antibodies to RNA polymerase
• -- Systemic sclerosis

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Antinuclear antibodies (ANAs)

• Peripheral (rim)
• Anti-ds-DNA SLE

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Antinuclear antibodies (ANAs)

• Centromere pattern
• Anti-centromere antibodies
• -- CREST syndrome, Sjögrens syndrome
• CREST syndrome Calcinosis, Raynauds phenomenon
  , Esophageal dysmotility, Sclerodactyly,
  Telangiectasia

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Antinuclear antibodies (ANAs)

• Cytoplasmic pattern
• -- may be related to primary biliary cirrhosis
  (PBC)

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Antineutrophil cytoplasmic antibodies (ANCA)

• c-ANCA
• cytoplasmic staining
• ??proteinase 3
• Wegener's Granulomatosis (90)
• Microscopic polyarteritis

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Antineutrophil cytoplasmic antibodies (ANCA)

• p-ANCA
• peri-nuclear stain
• against myeloperoxidase and against elastase
• seen in patient with GN, various forms of
  vasculitis and collagen vascular disease

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Anti-ds DNA Antibody

• ??DNA??????????,??DNA??(anti-ds DNA)??SLE????????
• 50-80 of SLE patients
• In SLE, anti-ds DNA antibody strongly correlate
  with nephritis and disease activity

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Anti-phospolipid antibodies

• Lupus anticoagulant, Anticardiolipin Ab,
  Anti-beta2-glycoprotein-I Ab, Antiphosphotidyl
  serine Ab (APTS)
• Thromboembolism events CVA, AMI, Pulmonary
  embolism, DVT, Ischemic bowel
• Recurrent abortion, Infertility, Pre-eclampsia
• Hemolytic anemia, thrombocytopenia
• SLE CNS involvement, Mononeuropathy multiplex
• Raynauds phenomenon, Livedo reticularis

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Anti-Ro La Antibody

• 30-40 of SLE patients possess anti-Ro activity
• 0-15 of SLE possess anti-La
• SS-B/La????????????Sjogrens syndrome???,???SS-B/L
  a?????,????SS-A/Ro??,??????

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Anti-Ro La Antibody

• Anti-Ro is associated with photosensitive rash,
  subacute cutaneous lupus and the neonatal lupus
  syndrome
• In SLE, anti-La correlates with late-onset SLE,
  secondary Sjogren's syndrome, the neonatal lupus
  syndrome

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Anti-Ro La Antibody

• Anti-Ro (SS-A) antibodies recognize a set of
  primarily cytoplasmic ribonucleoproteins
  consisting of the 60-kDa Ro60 antigen and a
  single molecule of human Y1, Y3, Y4, or Y5 RNA
• The 47-kDa La (SS-B) antigen associates
  transiently with the precursors of several small
  RNAs synthesized by RNA polymerase III, including
  Ro60 ribonucleoproteins, and protects them from
  exonucleases.
• La is a termination factor for RNA polymerase III

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Anti-Sm antibody

• Specific antibodies in the diagnosis of SLE
• 10 to 25 of SLE patients
• Anti-Sm and anti-RNP autoantibodies recognize a
  set of small nuclear ribonucleoproteins (snRNPs)
  containing highly structured, uridine-rich small
  RNAs (U1, U2, U4, U5, and U6 RNAs) involved in
  the splicing of messenger RNA
• Sm?RNP???mRNA????????Splicesome??,????RNA(SnRNA)??
  ?????????????????????RNA?

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Anti-Sm antibody

• Sm?RNP??/???????????????????????,??Sn RNA-??????
  U1?U2?U4/U6?U5???
• RNP????U1RNP ????(immuno precipitation)??,?Sm?????
  ??SnRNP???????20-30?SLE???Sm??,??????????SLE????
  ?

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Anti-RNP antibody

• 100 in MCTD?high-titer
• 26 to 45 of SLE
• lower titers in several other rheumatic diseases,
  including primary Raynauds phenomenon,
  rheumatoid arthritis, and scleroderma

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Nucleic acid components of some autoantigens
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Anti- histone antibody

• Histone?????,?DNA??,?????????H1?H2A?H2B?H3?H4
• In the 20 to 60 of SLE patients, predominantly
  target the H1 and H2B proteins
• appear uniformly in drug-induced lupus
• low frequency in other diseases

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Anti- Jo-1 antibody

• 20 to 50 of polymyositis
• ?????????????????,??????(GOT GPT?CPK???,??????????
  ?)???
• Jo-1??????????????????????,?Mi-2????????????,?Jo-1
  Ab????????????

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Anti- Scl 70 antibody

• Anti- Scl 70 antibody???topoisomerase I
• topoisomerse?????????(super-coiled)???DNA??,?????1
  00000,?????????????,??????????70KD??????Scl-70,???
  ???Scleroderma????,????????

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Autoantibodies in scleroderma

• Anti-centromere antibody limited cutaneous
  disease
• Anti-Scl-70 antibody diffuse skin disease and
  interstitial lung disease
• Anti-RNA polymerase III antibody worse
  prognosis, extensive skin thickening and renal
  crisis

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C3

• Decreased C3 levels associated with most active
  diseases with immune complex formation

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C3

• Decreased C3 levels
• Active SLE
• Absence of C3b inactivator factor
• Acute post-streptococcal glomerulonephritis
• Membranoproliferative glomerulonephritis
• End-stage liver disease
• Increased levels found in numerous inflammatory
  states.

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C4

• Decreased C4 levels
• Acute SLE
• Early glomerulonephritis
• Cryoglobulinemia
• Inborn C4 deficiency
• Hereditary angioneurotic edema
• Increased levels found in numerous inflammatory
  states.

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• ??????????????????
• Anti-dsDNA lupus nephritis (84.4)
• Anti-Ro/SSA - congenital heart block
• Anti-Ribosomal P psychosis (88.9)
  (neuropsychiatric)
• Anti-Neuronal seizure
• Anti-Sm specificity (?????) (87.1 85.7 84.10)
• (Anti-dsDNA?)
• Anti-phospholipid Ab--spontaneous abortion,
  neuropsychiastric lesions (CNS involvement) (88.9)

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• ??????????????????
• Anti-phospholipid antibody--Libman-Sachs?????(Libm
  an-Sachs endocarditis) (88.6)
• Anti-Ro/SSA- ???????????(subacute cutaneous lupus
  erythematosus) --???????????,????,??????????????
  (88.7)
• Anti-histone Ab drug-induced lupus (87.2)
  (86.3) (85.3)

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Rheumatoid Factor

• an autoantibody that recognize abnormally
  glycosylated IgG
• seropositive or seronegative diseases sero
  means rheumatoid factor
• RF???????IgG?IgM??,?????IgG?IgA?IgD?IgE?

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Rheumatoid factor in rheumatoid arthritis

• ??RA?criteria????
• High titers of RF, particularly IgG RF, are a
  risk factor for the development of vasculitis
• elevated IgA RFs may correlate with bone
  erosions, vasculitis, and a more severe disease
  course

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Anti-citrullinated protein antibodies (ACPA) or
anti-cyclic citrullinated protein antibodies
(anti-CCP)

• During inflammation, arginine residues in
  proteins can be enzymatically converted into
  citrulline ones (a process called
  citrullination), and, if their shapes are
  significantly altered, the proteins may be seen
  as antigens by the immune system
• The most specific marker for the diagnosis of
  rheumatoid arthritis

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Anti-citrullinated protein antibodies (ACPA) or
anti-cyclic citrullinated protein antibodies
(anti-CCP)