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Fluorescence patterns of nuclear autoantigens

Autoantibodies recognizing chromatin related antigens


chromatin image


Chromatins are composed of dsDNA and its binding proteins (histones and non-histone proteins). 146 base pairs of dsDNA wind around histone core (histone octamer composed of 2 molecules each of H2A, H2B, H3 and H4 subunits) to make a nucleosomal unit which connect to each other through a DNA linker with histone H1 subunit. In interphase cells, chromatins are dispersed in the nucleoplasm and condensed to form chromatids (chromosomes) in mitotic cells.


Anti-dsDNA antibody


photo 13
Peripheral staining

photo 14    AF/CDC-1*
Homogeneous staining


photo 15
kinetoplast of Clithidia luciliae is stained.


Pattern Peripheral and homogeneous staining in interphase cells with strong chromosomal staining in mitotic cells. At lower serum dilutions (x 20-x 40), the antibodies tend to give peripheral patterns and often show homogeneous staining at higher serum dilutions. Peripheral patterns are more frequently found when rodent tissue sections are used as substrates.
Antigen Double stranded DNA(dsDNA). Anti-ssDNA antibodies do not give fluorescence.
Other analytical
method
IIF, ELISA, RIA, CLEIA
Clinical
significance
The antibody is found flequently and specifically in SLE patients with active renal disease. The presence of Anti-dsDNA is a major criteria for the classification of SLE.
References 5, 6

* Reference serum from Centers for Disease Control



Anti-DNA histone complex antibody (anti-DNP antibody / anti-nucleosomal antibody)


photo 16


Pattern Homogeneous nuclear staining in interphase cells and intense fluorescence in the chromosomal region in mitotic cells.
Antigen Histone-DNA complex with the antigenic epitope in DNA-histone binding sites.
Other analytical
method
ELISA
Clinical
significance
SLE, lupus nephritis
References 7, 8



Anti-histone antibody


Pattern Similar to the former antibody
Antigen Histone H1, H2A, H2B, H3, H4, H2A-H2B complex (major antigen in IIF method) and H3-H4 complex.
Other analytical
method
ELISA, WB
Clinical
significance
SLE (antibodies to H1 and H2A), drug-induced lupus (antibodies to H2A-H2B complex)
References 9



Anti-Ku antibody


photo 17


Pattern Speckled (patchy) nuclear staining (sometimes described as homogeneous staining) with no chromosomal staining in mitotic cells.
Monoclonal antibodies to the 70kD subunit give similar patterns.
Antigen DNA binding 80kD/70kD heterodimer protein (DNA-dependent protein kinase activator). Antibodies targeting either subunit or the complex are reported.
Other analytical
method
DID, Immunoprecipitation
Clinical
significance
SLE, SSc (USA)
polymyositis/scleroderma overlap syndrome (Japan)
References 1, 4, 10, 11



Anti-Scl-70 antibody (anti-Topoisomerase I antibody)


photo 18
AF/CDC-9*


Pattern Homogeneous (dense fine speckled to homogeneous) staining with chromosomal staining in interphase cells.
Nucleolar staining is often observed.
Antigen DNA Topoisomerase I
Other analytical
method
DID, ELISA, WB, CLEIA
Clinical
significance
Antibodies to topoisomerase I are generally found in systemic sclerosis with high sensitivity and their presence constitutes one of the diagnostic criteria of the disease.
References 12



Anti-centromere antibody


photo 19
AF/CDC-8*


photo 20
2 fluorescent dots are visible for each chromosomal pair. (hypotonic spread preparation of mitotic chromosomes)

Pattern Centromere or discrete speckled staining. 40-80 fine speckles are stained in the nucleus of interphase cells. The speckles are characteristically aligned at the chromosomal region in mitotic cells. Paired centromeric fluorescence staining is observed in the chromosomal spread preparations.
Antigen Centromere proteins; CENP-B (80kD), CENP-A (17kD), CENP-C (140kD)
Other analytical
method
ELISA, WB, CLEIA
Clinical
significance
These antibodies are consistently found in patients with the limited form of SSc and their presence constitutes a diagnostic criteria of SSc. These antibodies are also found in PBC.
References 13, 14


centromere image


The constricted region where two sister chromatids join along is the centromere. CENP is a specific protein in this region. Microtubules of the spindle apparatus attach to kinetochore when the chromatid separation occurs.


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SLE: systemic lupus erythematosus, SSc: systemic sclerosis, MCTD: mixed connective tissue disease, SS: Sjögren's syndrome, PM/DM: polymyositis/dermatomyositis, RA: rheumatoid arthritis, PBC: primary biliary cirrhosis, AIH: autoimmune hepatitis


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