Anti DsDNA Antibodies
Anti DsDNA Antibodies
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AUTOIMMUNITY
Autoantibody
Autoantigen
SLE
prevalence
Correlation
Clinical
with Disease Associations
Activity
Antinuclear antibodies
(ANA)
Anti-Deoxyribonucleic
acid (DNA)
>95%
No
40-80%
(depending on
assay type)
Yes
Anti-Sm antibody
Comprised of at least 8
polypeptides in the SmsnRNP complex
30-40%
Yes
Anti-nucleosome
antibody
50-90%
Yes
Anti-anionic
phospholipid antibodies
Comprised of DNA
wrapped around core
histones
Mostly anti-cardiolipin
(aCL)
aCL 21-53%
Yes
Anti-Beta2-glycoprotein
1 antibodies
Anti-C1q antibodies
17-49%
Debated
30-50%
Correlate with
nephritis activity
inflammation. Renal, neurological, and haematological symptoms are three further criteria. The
two remaining ACR criteria relate to abnormalities
in the levels of serum autoantibodies [Table 1].Any
patient who satisfies, either simultaneously or serially, four out of these eleven ACR criteria during
any period of observation is classified as having
SLE [1]. There are several recognised methods for
the assessment of disease activity in SLE. These
include the ACRs systemic lupus erythematosus
disease activity index (SLEDAI) and the predominantly UK-based British Isles Lupus Assessment
Group (BILAG).
Autoantibodies
Reported autoantibody targets in SLE include
nuclear and cytoplasmic macromolecules, lipid
components and plasma proteins. Table 2 presents
some of the antigenic specificities of autoantibodies found in SLE patients. The most frequently
associated autoantibody specificities include Sm,
nucleosomes, histones and double-stranded DNA
(dsDNA). Anti-dsDNA autoantibodies are the
most frequently detected.
ANA
Elevation of anti-nuclear antibodies (ANA) is one
of the most sensitive serological ACR criteria. More
than 95% of patients with SLE have an elevated
ANA titre at some point during the course of their
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AUTOIMMUNITY
lupus nephritis between disease activity and antidsDNA antibody levels. Notably, rising titres of
antibodies to dsDNA may indicate exacerbations
of glomerulonephritis [6].
Auto-antibodies
against double
stranded DNA
are those most
frequently
detected in SLE.
The presence
of such
anti-dsDNA
antibodies is
one ACR
criterion for
the diagnosis
of the disease.
Anti-dsDNA antibodies
First described in the late 1950s autoantibodies to
deoxyribonucleic acid (DNA) are highly heterogeneous with respect to their avidity, immunoglobulin subclass composition, cross-reactivity and complement fixing ability. There is also some debate as
to whether dsDNA is always the principle antigen
for anti-dsDNA antibodies in SLE; nucleosomes
should also be considered as relevant antibody targets. Native DNA exists primarily as a double
stranded right handed helix (dsDNA) and is frequently found in association with histones, in the
form of nucleosomes.
In SLE, anti-DNA antibodies are classified according to their reactivity to dsDNA; antibodies to single stranded DNA (ssDNA) are not specific for SLE
as they are found in sera from patients with both
Figure 1. The principle behind RIA method for the determination of antibodies to dsDNA in serum.
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Methodology
Summary
of method
Farr RIA
IFA Crithidia
ELISA
AUTOIMMUNITY
Avidity of
antibody
detected
High
Class of
antibody
detected
All
Advantage
Result format
Disadvantage
High specificity
for SLE
Quantitative
Can be
standardised in
IU/mL
Medium
to high
Dependent
on conjugate
specificity
Semi-quantitative
result - either
positive, negative
or endpoint titre
High and
low
Most
commonly
IgG class
1. Specific
2. Common
laboratory
technique
3. Isotype of
antibodies can
be determined
4. No
interference
from antibodies
to ssDNA
Highly Sensitive
Automation
friendly.
Avoids the use
of radiolabels
Quantitative
results IU/mL
The use of ELISAs for the assay of dsDNA antibodies is well established. In addition to giving a
quantitative result, the ELISA method is easy to
perform, relatively inexpensive and does not
involve the use of any radiolabels. ELISAs are
readily standardised using the World Health
Organisation (WHO) reference preparation for
anti-dsDNA, Wo80. It is important to confirm
that the ELISA does not detect anti-ssDNA antibodies which are not specific for SLE. This can be
easily verified by including an anti-ssDNA sample amongst the assay controls.
Other Methods
More recent methods for detecting anti-dsDNA
antibodies include techniques such as immunoblotting, bead-based immunoassays and automated
Farrzyme positive
Farrzyme negative
Farrzyme borderline
Figure 3. The relative avidity of 23 samples was determined by competition assays. The above graph shows the
results from these samples using the new Farrzyme assay. It can be seen that the new assay correctly
identifies high avidity samples.
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For an individual patient with SLE the antidsDNA antibody avidity tends to remain more or
less constant over time; exceptions are found
among patients who develop nephritis during
the course of the disease. Those SLE patients initially found to have only lower avidity anti-DNA
antibodies may have a milder form of SLE with
less frequent episodes of nephritis [3, 6].
References
1. Feletar M et al. The impact of the 1997 update of the
American College of Rheumatology revised criteria
for the classification of systemic lupus erythematosus:
what has been changed? Arthritis Rheum 2003;
48(7):2067-9.
2. Hochberg MC. Updating the American College of
Rheumatology revised criteria for the classification of
systemic lupus erythmatosus. Arthritis Rheum 1997;
40: 1725.
3. Isenberg D., Smeenk R. Clinical laboratory assays for
measuring anti-dsDNA antibodies. Where are we
now? Lupus 2002; 11: 797-800.
4. Jaekel HP, Trabandt A et al. Anti-dsDNA antibody
subtypes and anti-C1q antibodies:toward a more
reliable diagnosis and monitoring of systemic
lupus erythematosus and lupus nephritis. Lupus
2006; 15: 335-45.
The author
Richard Hughes, Ph.D., & Sarea UI-Hassan
The Binding Site Ltd,
Birmingham,
UK.
Tel +44 121 436 1000
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Conclusion
The detection of abnormal titres of anti-dsDNA
antibodies is a valuable tool for the clinician,
both as a diagnostic marker and to monitor disease activity in SLE. Studies have indicated a correlation between disease activity and high avidity
anti-dsDNA antibodies in lupus nephritis. The
three most common assay methods namely
ELISA, Crithidia luciliae IFA, and Farr RIA each
detect a different spectrum of anti-dsDNA antibodies. Only the Farr RIA is selective for those
antibodies of higher avidity, but this assay is
References:
1. Isenberg D., Smeenk R. Clinical laboratory assays for measuring anti-dsDNA
antibodies. Where are we now? Lupus 2002;11:797-800
2. Jaekel H.P. et al. Anti-dsDNA antibody subtypes and anti-C1q antibodies:
toward a more reliable diagnosis and monitoring of systemic lupus erythematosus
and lupus nephritis. Lupus 2006;15:335-45
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