The effects of affinity-purified anti-DNA antibodies from patients with systemic lupus erythematosus on the fluorescent antinuclear antibody assay using HEp-2 cells.
The aim of this study was to clarify the effects of anti-dsDNA antibodies on the titer and the nuclear staining pattern(s) in a fluorescent antinuclear antibody (FANA) assay using HEp-2 cells. Anti-dsDNA derived from 14 patients with systemic lupus erythematosus (SLE) was individually affinity-purified. The anti-dsDNA titer of the purified anti-dsDNA solution was measured by radioimmunoassay (RIA) or by enzyme-linked immunosorbent assay (ELISA). In the FANA assay, the anti-dsDNA solution was diluted in a stepwise manner and its titer was expressed by the endpoint dilution. The nuclear staining pattern in the anti-dsDNA solution was examined at the 1:5 and 1:20 dilutions and at the endpoint dilution. The anti-dsDNA titers of the affinity-purified anti-dsDNA solution were high enough (13 to 126 IU/ml) to be measured by RIA. However, the antinuclear antibody (ANA) titers of this solution were relatively low: 1:20 to 1:320. In the study of nuclear staining the peripheral pattern was observed in nine of the 14 cases at a 1:5 dilution. However, at the endpoint dilution, all cases exhibited the homogeneous pattern. These findings indicate that in the FANA assay using HEp-2 cells, 1) although serum samples show high anti-dsDNA titers by RIA or by ELISA, the antibodies' direct contribution to ANA titers is limited, and 2) when samples reveal a homogeneous staining pattern at the endpoint dilution, this suggests the presence of anti-dsDNA.
Suzuki
Kimihiro
K
Internal Medicine I, National Defense Medical College, Tokorozawa, Japan. kogen@me.ndmc.ac.jp
Kawamura
Masahide
M
Mineo
Midori
M
Shinohara
Tadashi
T
Kataharada
Koji
K
Okada
Makoto
M
Takada
Kunio
K
Miyawaki
Shoji
S
Ohsuzu
Fumitaka
F
D000327:Adsorption; D000328:Adult; D000368:Aged; D000974:Antibodies, Antinuclear / Q000276:immunology* / Q000302:isolation & purification*; D004247:DNA / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D008875:Middle Aged; D011863:Radioimmunoassay; D014407:Tumor Cells, Cultured
D000974:Antibodies, Antinuclear; D004247:DNA