Reference sera for antinuclear antibodies. I. Antibodies to native DNA, Sm, nuclear RNP, and SS‐B/La

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Objective. To define the fine specificity of the 10 reference sera used for determination of antinuclear antibodies (ANA) and ANA subsets which are available from the Arthritis Foundation (AF) and from the Centers for Disease Control and Prevention (CDC). Methods. AF/CDC sera were assessed by experienced laboratory staff, using indirect immunofluorescence and Western blotting. Results. The original assignment of fluorescence patterns to 4 reference sera was confirmed, and the fluorescence intensities were determined using reference fluorescent beads. On Western blots, sera AF/CDC2 (anti—SS‐B/La) and AF/CDC7 (anti—SS‐A/Ro) did not detect Ro antigens, sera AF/CDC9 and AF/CDC10 appeared to be monospecific anti—Scl‐70 and anti—Jo‐1 sera, respectively, serum AF/CDC4 (anti—U1 small nuclear RNP) recognized the 70‐kd band, and serum AF/CDC5 recognized the Sm antigen with its multiple bands. Semiquantitative analyses revealed that AF/CDC5, AF/CDC2, and AF/CDC10 were strongly reactive sera, whereas AF/CDC4 and AF/CDC9 were much weaker and should be used at lower dilutions on Western blots. Conclusion. The AF/CDC ANA reference sera, originally described as reference reagents for indirect immunofluorescence and double immunodiffusion techniques, are also useful for Western blotting. The data presented herein further support the use of these sera for reference purposes.

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Reference sera for antinuclear antibodies. II. Further definition of antibody specificities in international antinuclear antibody reference sera by immunofluorescence and western blotting

Smolen

Butcher

Fritzler

et al. 1997

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“…The presence of precipitating antibodies to Sm, RNP, SS-A, and SS-B was examined by Ouchterlony double diffusion, using rabbit thymus and human spleen extracts at protein concentrations of 100 mg/ml and 40 mg/ml, respectively. Known positive serum controls were provided by the CDC (20). Antibodies to native DNA were measured by the Farr assay, using phage h 3H-DNA, radiolabeled in vitro by nicktranslation (21).…”

Section: Methodsmentioning

confidence: 99%

Airways disease in rheumatoid arthritis patients

Radoux

Méanard

Béagin

et al. 1987

Airflow limitation is a frequent finding in patients with rheumatic diseases. We have previously suggested that it is associated with autoimmune exocrinopathy in Sjögren's syndrome. To compare clinical features of patients with and without airways dysfunction and to further test the hypothesis of a link between airways disease and exocrinopathy, we prospectively studied 2 groups of 15 lifetime nonsmoker female patients with seropositive rheumatoid arthritis (RA). The 2 groups were similar in their clinical and immunologic features, but differed in terms of airways function. Salivary, lacrimal, and sweat gland dysfunction were significantly more prevalent or severe in the group with airways disease. Antinuclear antibodies were also more prominent in the patients with airways disease, but antibodies against RNP, SS‐A, SS‐B, and double‐stranded DNA were not present in these patients. HLA‐DR4 was found in 80% of the RA patients with airways disease and in 57% of those without airways disease. HLA–B8 and DR3 were equivalently distributed in both groups. This prospective study further documents the existence of small airways disease in RA and supports the view that autoimmune exocrinopathy predisposes to its expression.

“…The Sm, U1 RNP, SS-B (La), rheumatoid arthritis nuclear antigen (RANA), and SS-A (Ro) prototype sera have been previously described and have been used as standards itl this laboratory for several years (7). Standards for anti-Sm, anti-U1 RNP, and anti-SS-B (La), obtained from the Centers for Disease Control (Atlanta, GrA) were also used to corroborate specificity (8).…”

Section: Methodsmentioning

confidence: 99%

IgM anti‐histone H‐3 antibody associated with undifferentiated rheumatic disease syndromes

Molden¹,

Klipple²,

Peebles³

et al. 1986

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A distinctive type of speckled antinuclear antibody staining pattern was identified by indirect immunofluorescence on mouse kidney substrate in 4.8% of 5,976 specimens analyzed for antinuclear antibodies. This pattern, termed variable large speckles (VLS), consisted of 3-10 nuclear speckles ranging in size from approximately 0.2-2.0~. The pattern could be differentiated from other indirect immunofluorescence patterns related to specific antibodies. The predominant immunoglobulin isotype demonstrating the VLS pattern was IgM in 27 of 28 sera examined and IgG in 1 serum. VLS sera had substantial IgM antibodies to histone demonstrated by enzyme immunoassay , and further analysis of representative sera showed predominant antibody activity to histone class 3 (H-3). Adsorption with histone H-3 resulted in decrease or removal of antibody producing the VLS pattern. Available informatiofi showed that most patients with LgM antibodies of the VLS pattern had undifferentiated connective tissue disease symptoms. They were characterized by a heterogeneity of chronic symptoms including arthralgias, myalgias, inflammatory polyarthritis, myo- The indirect immunofluorescence technique is a widely used method of screening for the presence of antinuclear antibodies (ANA). The patterns of nuclear fluorescence observed with this assay are indicative, but not diagnostic, of a specific type of antibody (1). There are many ANAs which display a variety of speckled staining patterns; among these are antibodies to Sm, RNP, SS-A (Ro), and SS-B (La). A speckled pattern which can be distinguished from others is that of the antibody to centromere/kinetochore (2). We report here another distinctive type of speckled pattern which is different from other speckled patterns and was first observed when mouse kidney sections were used as the substrate for the indirect immunofluorescence test. It consists of 3-10 distinct, variably shaped, 0.2-2.0~ nuclear speckles and will be referred to as the variable large speckled (VLS) antinuclear antibody pattern. Our studies provide information on the frequency, the clinical significance, the substrate, and the immune specificity of the VLS pattern. PATIENTS AND METHODSPatient sera. Of the 5,976 sera sent to this laboratory for routine ANA testing, using mouse kidney sections as substrate, 289 (4.8%) were observed to have multiple, variably sized fluorescent nuclear speckles. They were classified as having a "variable speckled" ANA pattern. Titration data were available on 210 of these 289 sera, and 81 of the 210