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Fluorescence patterns of nuclear autoantigens

Autoantibodies recognizing
cell cycle related antigens



The remarkable differences in the patterns and intensities of nuclear staining among HEp-2 cells on the slide indicate the presence of autoantibodies targeting the antigens which show changes in their quantities, localizations and molecular structures dependent on the cell cycle. Sometimes, coexistence of more than two autoantibodies is suggested by the pleomorphic staining patterns. However the details are not clear.


Anti-PCNA antibody


photo 35


Pattern Most intense nuclear staining occur in S phase cells. The antibodies are characterized by the variation of staining patterns in the cells in different cell cycles; sparse fine speckled nuclear staining in late G1 phase cells, dense homogeneous staining in S phase cells, and coarse nuclear staining with no nucleolar decoration in late S phase cells. The chromosomal regions show no staining in mitotic cells. When the antibodies coexist with other antibodies, it is sometimes useful to confirm the specific staining by serial serum dilutions.
Antigen 34kD auxiliary protein of DNA polymerase δ. Recently, it was reported that PCNA antigen forms a macromolecule complex with Ki antigen.
Other analytical
method
DID, WB
Clinical
significance
These antibodies occur in patients with SLE (1-2%), but rarely seen in other diseases.
References 36, 37, 38


Anti-Na antibody


photo 36


photo 37

Pattern Nuclear speckled staining is seen mainly in G2 phase cells, but cell cycle-dependent polymorphic staining is not remarkable compared with that of anti-PCNA antibody. The chromosomal regions of mitotic cells show speckled staining similar to those given by anti-centromere antibody. The cleavage furrow observed in the telophase of mitosis and the midbody after cell division is stained by the antibodies.
Antigen Myosin related antigen
Clinical
significance
The presence of this antibodies was reported in patients with SLE.
References 39, 40

Anti-CENP-F antibody was reported to give similar staining patterns to anti-Na antibody. They occur in malignant thoracic tumors such as lung cancer and recognize one of the centromere proteins (CENP-F 367kD). [References:44, 73]
Anti-midbody (MSA-2) antibody occuring in scleroderma show negative or very weak nuclear staining in interphase cells compared with anti-Na antibody. [References: 43, 44]
Anti-MSA-3 antibody shows speckled staining in some of the interphase cells, but do not stain the cleavage furrow in telophase cells. The antibodies were reported to occur in association with cancer of the respiratory system. [Reference: 44]
It was also reported that anti-DNA topoisomerase II antibodies are observed in hepatocellular carcinoma. [Reference: 30]



The photos shown below are staining patterns of the antibodies recognizing cell cycle related antigens, observed in MBL Co., Ltd.. Details of the antigens and the clinical significance are not determined.



photo 38
Homogeneous nuclear staining in some of the interphase cells with more intensive staining of chromosomal region in mitotic cells.


photo 39
Weak nuclear fluorescence in some of the cells with cytoplasmic staining.


photo 40
Some of the nucleoli are stained.


photo 41
No nuclear staining in interphase cells with intense chromosomal staining in mitotic cells.


Many proteins have been found to be associated with proliferation of the cells. Among them are some of the antinuclear antibodies with cell cycle dependency. The autoantibodies observed in patient sera may provide useful tools for studying the structure and function of these autoantigens. [Reference: 45]


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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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