Anti-Nuclear Antibody (ANA) Anti-Nuclear Antibody (ANA)
1. Screening Test E. High titers (>1:320) is highly suggestive of an
A. Refers to auto-antibodies to nuclear autoimmune disease
components of the cell F. Low or negative result does not rule out
B. Seen in a wide variety of systemic autoimmune autoimmune disease
diseases G. Many patients with different kinds of
C. Useful as diagnostic markers and to monitor autoimmune diseases will have generic ANA’s
the course of autoimmune diseases H. Specific types of ANA’s will help to identify a
D. Can be found in low titers in healthy specific autoimmune diseases:
individuals
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Anti-Nuclear Antibody (ANA) Anti-Nuclear Antibody (ANA)
1. Incidence of ANA in Various
I. ANA Screening Tests Diseases of Conditions Disease % Positive
1) Indirect Immunofluorescence (See Table to Right) Systemic Lupus Erythematosus (SLE) Over 95%
Progressive Systemic Sclerosis (Scleroderma) 60-90%
a) Hep-2 cell substrate 2. ANA’s stain different nuclear
components
1. Human cancer cell line a. ds-DNA
Rheumatoid Arthritis 25-30%
b) Incubate patient serum on cell substrate b. Histones
Sjogren’s Syndrome 40-70%
c. Non-histone proteins
c) Wash d. Nucleolus
Felty’s Syndrome 1005
d) Incubate substrate with fluorescent-conjugated
3. There are 5 major staining
anti-human antibody patterns Juvenile Arthritis 15-30%
a. Homogeneous
e) Observe staining pattern under fluorescent b. Rim
microscope c. Speckled
d. Nucleolar
e. Centromere
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Anti-Nuclear Antibody (ANA) Anti-Nuclear Antibody (ANA)
Homogeneous Pattern Rim Pattern
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CLS 420 Clinical Immunology & Molecular Diagnostics – Lecture Series
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Anti-Nuclear Antibody (ANA) Anti-Nuclear Antibody (ANA)
Speckled Pattern Nucleolar Pattern
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Anti-Nuclear Antibody (ANA) Anti-Nuclear Antibody (ANA)
Centromere Pattern Summary
Resting Cell Dividing Cell
The centromere pattern has the following Rim Pattern Probably anti-dsDNA. Your patient probably has
characteristics: systemic lupus
The resting cell nuclei have numerous
discrete speckles. If it were possible to count Homogeneous Pattern Probably anti-histones. Your patient probably has
them most cells would have 46 dots, the
drug-induced lupus
number of chromosomes present in human
cells.
The chromatin of the dividing cells has
Speckled Pattern Could be anti-Sm and/or anti-Ro/SSA and/or anti-
speckled staining. La/SSB and/or anti-U1RNP and/or any of several
others. You'll certainly want to continue your
workup
Discrete
Speckled
Centromere Pattern An especially fine speckling with little background
Staining staining. This is anti-centromere, the marker for
CREST/Pulmonary hypertension
Speckled Nucleolar Pattern anti-Th or anti-fibrillarin / anti-U3RNP or anti-
Staining of
the U17RNP. Think of scleroderma, though most
Chromatin scleroderma patients don't show the nucleolar
pattern
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Anti-Nuclear Antibody (ANA) Anti-Nuclear Antibody (ANA)
Anti-dsDNA Antibodies Anti-dsDNA Antibodies
K. Specific Information About Follow-up Tests for a Positive ANA
J. Follow-up Tests for a Positive ANA Screen Screen
1) Anti-double-stranded DNA (dsDNA) antibody 1) Anti-dsDNA Antibodies
2) Anti-Smith (Sm) antibody a) Peripheral or homogeneous pattern on routine IFA
3) Anti-histone antibody b) Definitive detection and quantitation is performed on
4) Anti-ribonucleoprotein (RNP) antibody more specific substrates
5) Anti-Sjogren’s Syndrome (SSA and SSB) antibody 1. IFA using the hemoflagellate Crithidia luciliae as
6) Anti-Scl-70 antibody substrate
7) Anti-centromere antibody 2. Very specific for SLE
8) Anti-extractable nuclear antigen (ENA) antibody 3. 40-60% of patients with SLE have anti-
dsDNA antibodies
9) Jo-1 (Histadyl tRNA Synthetase)
4. Tends to be a Rim Pattern
c) Positive results support a diagnosis for SLE and can be
used to monitor disease activity and response to therapy
1. Found in 40-60% of patients with SLE during the
active phase of their disease
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CLS 420 Clinical Immunology & Molecular Diagnostics – Lecture Series
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Anti-Nuclear Antibody (ANA)
Anti-Nuclear Antibody (ANA)
Crithidia Organisms (dsDNA)
2) Anti-Smith (Sm) antibodies
a) Speckled pattern on IFA
b) Specific for SLE
c) Only 30% of patients with SLE have high
titers
d) Used in follow-up to collagen vascular
disorders
3) Anti-Histone antibodies
a) Homogeneous pattern on IFA
b) Most commonly seen in drug-induced lupus
c) In drug-induced lupus there is not normally
n association with other autoantibodies
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Anti-Nuclear Antibody (ANA) Autoantibodies in Rheumatic Diseases
4) Anti-RNP antibodies (Speckled or Nucleolar Pattern)
a) Specifically for the differential diagnosis of: Disease Antibody to:
1. Systemic rheumatic diseases SLE dsDNA and/or Sm
2. Follow-up analysis of collagen vascular disorders
5) Anti-SSA and Anti-SSB (Speckled Pattern) Drug induced SLE Histone
a) Valuable when a patient is anti-nuclear antibody SCLE Anti Ro (SS-A)
negative but thought to have SLE
b) Strongly associated with Sjogren’s Syndrome MCTD RNP
6) Anti-SCL-70 antibodies (Speckled Pattern) Scleroderma (Diffuse) Scl-70
a) Seen in about 20% of patients with diffuse scleroderma
CREST Centromere
b) Highly specific for diffuse scleroderma
7) Anti-Centromere antibody appears in 90% of patients with Polymyositis/Dermatomyositis Jo-1 (histadyl tRNA synthetase)
CREST syndrome which is a variant of scleroderma
Sjogren’s Syndrome Anti Ro (SS-A), SS-B
8) Anti-ENA antibodies (Speckled Pattern)
a) Systemic rheumatic diseases Wegener’s Granulomatosis cANCA (Proteinase 3)
9) Jo-1 (Histadyl tRNA Synthetase)
a) Polymyositis/Dermatomyositis
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Anti-Nuclear Antibody (ANA) Anti-Nuclear Antibody (ANA)
Pattern Cell Target Disease Association Pattern Cell Target Disease Association
Nuclear Cytoplasmic
SLE (95%) Fine Speckled Jo-1 Polymyositis (20-40%)
Discoid Lupus
dsDNA
Autoimmune Hepatitis Ribosomal Pattern Ribosome P SLE (10-15%)
Homogeneous Rheumatoid Arthritis
Primary Biliary Cirrhosis (90%)
Histones SLE Mitochondrial M2
Scleroderma (40%)
Extensive Scleroderma (30%) ER CytochromeP450 Drug Induced Hepatitis
Nucleolar PM-Scl Polymyositis/Scleroderma overlap syndrome
Primary Pulmonary Hypertension SLE (rare)
Golgi Various
Sjogren's Syndrome
Mixed Connective Tissue Disease Rheumatoid Arthritis (30-80%)
Nuclear Matrix hRNP
SLE Mixed Connective Tissue Disease
Sjogren's Syndrome (95%)
Cytokeratin Cytokeratin Autoimmune Hepatitis
SS-A (Ro) Crohn's Disease
Speckled - fine SS-B (La)
SLE (40%) Squamous Cell Carcinoma of Lung
Scleroderma (5%)
SLE Contributed by Dr M Cobbold
Speckled - course snRNP
SLE overlap Syndromes This page is maintained by Clinical Immunology.
Limited Scleroderma / CREST (60%) Low titre ANAs (ie 1:40 or 1:100) are usually not clinically relevant whereas high titre ANA
Centromere CENT A-E
Raynauld's Phenomenon
(particularly 1:1600) are infrequent amongst healthy individuals and so therefore carry a
Proliferating Cell greater clinical significance. The table above are of common ANA patterns and there clinical
Nuclear Antigen Cyclin SLE (1-3%) - often associated with glomerulonephritis association. Possibly the most important use is to exclude SLE.
(PCNA)
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Nuclear Dots p80 coilin Autoimmune and viral liver disease
CLS 420 Clinical Immunology & Molecular Diagnostics – Lecture Series
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