The Use of Immunoblotting to Detect Antibodies to Nuclear and Cytoplasmic Antigens

Rooij, D. J. De; Putte, L B van de; Habets, W. J.; Verbeek, A.L.M.; Venrooij, Walther J. van

doi:10.3109/03009748809105271

Cited by 49 publications

(33 citation statements)

References 23 publications

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“…In an immunoprecipitation assay using the radiolabeled second and fourth stemloop as antigen (similar to experiments shown in Fig. 2 A), it then could be demonstrated in 10 (Fig. 3 B and C).…”

Section: Resultsmentioning

confidence: 99%

“…Most ofthe patient sera were obtained from the Rheumatology Department of the University Hospital Nijmegen, the Netherlands. All patients used in this study obtained the diagnosis SLE overlap syndrome or MCTD according to criteria described previously ( 10). Sera from these patients all contain anti-(U)RNP antibody as detected by both immunoblotting and counterimmunoelectrophoresis.…”

Section: Methodsmentioning

confidence: 99%

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Epitope regions on U1 small nuclear RNA recognized by anti-U1RNA-specific autoantibodies.

Hoet

Weerd²,

Gunnewiek³

et al. 1992

J. Clin. Invest.

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Autoantibodies specifically directed to U1RNA were found in patients suffering from systemic lupus erythematosus (SLE) overlap syndromes. To obtain more insight in the mechanism responsible for this UlRNA-specific antibody formation and to use the antibodies eventually as a tool to study UlRNA-protein (UlRNP) interactions, the B cell epitopes on U1RNA were mapped. Using in vitro synthesized domains of U1RNA, the main epitope regions were found in stemloops II and IV. Furthermore, 3'-end or 5'-end truncation of both stemloop II and stemloop IV showed that the conformation of the stemloops is critical for antibody recognition. Mutant studies on both stemloops indicated that in the case of stemloop II the stem is the main antigenic region, whereas in stemloop IV, the loop (Eloop) is a main target. The results of this study support the idea that the anti-U1RNA autoantibody could be the result of a process driven by the human U1RNP complex itself (antigendriven process). (J. Clin. Invest. 1992. 90:1753-1762

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Epitope regions on U1 small nuclear RNA recognized by anti-U1RNA-specific autoantibodies.

Hoet

Weerd²,

Gunnewiek³

et al. 1992

J. Clin. Invest.

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“…2 The availability of data on the frequency and levels of autoantibodies within a normal population is important for diagnosis. [1][2][3] Many such population studies have been performed in healthy individuals in various communities. [4][5][6][7][8][9] Oman occupies the southeastern corner of the Arabian Peninsula and has an estimated population of 2.4 million.…”

mentioning

confidence: 99%

“…Autoantibodies can be detected in sera of healthy individuals, but the literature clearly defines their role in specific autoimmune conditions. [1][2][3] The definition of abnormality related to autoantibody levels is usually based on abnormal quantity. Their presence merely supports but not confirms the diagnosis of an autoimmune disease.…”

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confidence: 99%

Frequency and Levels of Autoantibodies in Healthy Adult Omanis

2003

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Background: A previous pilot study showed a high frequency of anti-smooth muscle autoantibody in Omani blood donors and pregnant women. We conducted this larger-scale study to investigate the frequency and significance of several autoantibodies in healthy individuals from different regions of Oman. Methods: Sera obtained from 1537 healthy Omanis (1153 males and 384 females), ranging in age from 18 to 57 years, were tested for the presence of ten different autoantibodies using indirect immunofluorescence, haemagglutination and latex agglutination techniques. Results: Low levels of autoantibodies were detected in 33.5%, whereas a few individuals (1.8%) showed high autoantibody titres. Anti-smooth muscle autoantibodies (ASMA) were the most prevalent (11%). Anti-nuclear autoantibodies (ANA) were the second most prevalent (7.6%). Anti-thyroid microsomal autoantibodies (ATMA) and anti-thyroglobulin autoantibodies (ATA) were present in 6.5% and 4.4% of individuals, respectively. The other autoantibodies were detected much less frequently: anti-parietal cells autoantibodies (APCA) were found in 1.6%, anti-brush border antibodies (ABBA) in 1.3%, anti-reticulin autoantibodies (ARA) in 1%, antimitochondrial antibodies (AMA) in 0.8%, antiglomerular basement membrane antibodies (AGBMA) in 0.7% and rheumatoid factor (RF) in 0.4%. Conclusion:The data indicate that autoantibodies are present in healthy Omani individuals, and therefore caution should be taken when interpreting laboratory results of patients suspected of having autoimmune disease. Ann Saudi Med 23 (6):372-375

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“…The data on the frequency of a-Abs in healthy individuals is important for the diagnosis of autoimmune disorders 15,16,17,18 . Studies have been performed in healthy individuals in various countries 19 but limited data is available about the frequency of a-Abs in Pakistan, therefore, this study was designed to determine serum level of ANA, RA and anti dsDNA antibodies in apparently healthy population.…”

Section: Most Of the A-abs Belongs To Igg Class But Igm Andmentioning

confidence: 99%

Prevalence of Autoantibodies in Healthy Adults in Pakistani Population

Ashraf

Afzal

Kashif

et al. 2016

Bangladesh J Med Sci

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Introduction: Autoimmune phenomenon is attributed to a number of diseases which were once considered idiopathic. In humans, production of auto antibodies (a-Abs) against self-antigens is quite frequent but earlier their presence was associated with autoimmune diseases, however a-Abs have been documented in non-autoimmune disorders i.e. complicated pregnancy, cancer, stroke etc. In Pakistan, limited data is available on frequency of a-Abs, therefore this study was designed to determine serum level of antinuclear antibody (ANA), Rheumatoid factor (RF) and anti dsDNA antibodies in apparently healthy population. Materials and methods: After written informed consent, blood sample of 256 subjects was obtained by random sampling. Participants of established autoimmune diseases were excluded. Enzyme linked immunosorbent assay (ELISA) was used to determine ANA, RF and anti-dsDNA antibody. Categorical variables were compared by using χ 2 test. A p value <0.05 was considered statistically significant. Results: Rheumatoid factor was the most frequent a-Ab (18%), followed by anti dsDNA (7.4%), while ANA was the lowest (0.4%) antibody detected. Only RF had a statistically significant association with gender (p=0.047). No association of these antibodies with age was detected. Conclusion: Rheumatoid factor auto antibody was more prevalent as compared to ANA and dsDNA antibody in healthy adults.

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The Use of Immunoblotting to Detect Antibodies to Nuclear and Cytoplasmic Antigens

Cited by 49 publications

References 23 publications

Epitope regions on U1 small nuclear RNA recognized by anti-U1RNA-specific autoantibodies.

Epitope regions on U1 small nuclear RNA recognized by anti-U1RNA-specific autoantibodies.

Frequency and Levels of Autoantibodies in Healthy Adult Omanis

Prevalence of Autoantibodies in Healthy Adults in Pakistani Population

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