38 Molecular and Cell Biology of Autoantibodies and Autoimmunity
Titers of IgM RF and IgM antibodies to ssDNA were measured using an
ELISA procedure. Isolation of RF was carried out by incubation of serum with
human IgG-coupled Sepharose followed by elution with sodium acetate buffer
(pH 4.0).
The study disclosed that polyclonal RF cross-reactive with ss-DNA (CRRF)
are widely distributed in a variety of rheumatic diseases, and that their serum level
is significantly high in RA with extra-articular disease (EARA). CRRF would be
a useful marker for a clinical subset of RA. High prevalence rate of CRRF in
EARA may suggest its pathogenetic role in the extra-particular manifestation of
the disease, although the precise mechanism is not clear.
Profiles of Anti-Nuclear Antibodies in Scleroderma Spectrum
Disorders
H. Kondo, N. Takashina, and S. Kashiwazaki
Department of Internal Medicine, Kitasato University School of Medicine, Japan
In 112 scleroderma patients which satisfied the criteria for systemic sclerosis
of ARA and 43 patients with sclerodactyly and Raynaud’s phenomenon (SR),
anti-nuclear antibodies were studied. The presence of anti-centromere antibody
(ACA) was detected by staining of centromeres of chromosomes from Wil2 cells.
A nuclear enriched sonicate of HeLa cell proteins was prepared for immunoblot-
ting as described by McNeilage et al. to investigate antibodies to Ul-RNP,
Scl-70 and centromeric antigens. Furthermore, 17 kD centromeric antigen was
purified by immunoaffinity chromatography from the prepared crude antigens.
Immunoblotting was also performed using the purified 17 kD antigen.
Anti-nuclear antibody was detected by immunofluorescence using HEp-2 cell
in 95 of the 112 patients with scleroderma and in 36 of 43 with SR. Anti-Scl-70
antibody was detected in 32 of scleoderma sera and none of SR by immunodiffu-
sion. Thirty-one of 32 anti-Scl-70 antibody positive sera showed antibody for
95 kD protein by immunoblotting. Twenty-eight of scleroderma and 18 of SR had
precipitating antibodies to Ul-RNP. Twenty-six of 28 positive sera from scleroder-
ma and 13 of 18 positive sera from SR reacted with the 68 kD Ul-RNP associated
protein. Six of 7 sera which did not react with 68 kD protein recognized other
Ul-RNP associated proteins. Three patients (2.5%) with scleroderma had both
anti-Scl-70 and anti-Ul-RNP antibodies. ACA was detected in 17 of scleroderma
and 18 of SR by immunofluorescence. Antibody to 17 kD protein was demon-
Titers of IgM RF and IgM antibodies to ssDNA were measured using an
ELISA procedure. Isolation of RF was carried out by incubation of serum with
human IgG-coupled Sepharose followed by elution with sodium acetate buffer
(pH 4.0).
The study disclosed that polyclonal RF cross-reactive with ss-DNA (CRRF)
are widely distributed in a variety of rheumatic diseases, and that their serum level
is significantly high in RA with extra-articular disease (EARA). CRRF would be
a useful marker for a clinical subset of RA. High prevalence rate of CRRF in
EARA may suggest its pathogenetic role in the extra-particular manifestation of
the disease, although the precise mechanism is not clear.
Profiles of Anti-Nuclear Antibodies in Scleroderma Spectrum
Disorders
H. Kondo, N. Takashina, and S. Kashiwazaki
Department of Internal Medicine, Kitasato University School of Medicine, Japan
In 112 scleroderma patients which satisfied the criteria for systemic sclerosis
of ARA and 43 patients with sclerodactyly and Raynaud’s phenomenon (SR),
anti-nuclear antibodies were studied. The presence of anti-centromere antibody
(ACA) was detected by staining of centromeres of chromosomes from Wil2 cells.
A nuclear enriched sonicate of HeLa cell proteins was prepared for immunoblot-
ting as described by McNeilage et al. to investigate antibodies to Ul-RNP,
Scl-70 and centromeric antigens. Furthermore, 17 kD centromeric antigen was
purified by immunoaffinity chromatography from the prepared crude antigens.
Immunoblotting was also performed using the purified 17 kD antigen.
Anti-nuclear antibody was detected by immunofluorescence using HEp-2 cell
in 95 of the 112 patients with scleroderma and in 36 of 43 with SR. Anti-Scl-70
antibody was detected in 32 of scleoderma sera and none of SR by immunodiffu-
sion. Thirty-one of 32 anti-Scl-70 antibody positive sera showed antibody for
95 kD protein by immunoblotting. Twenty-eight of scleroderma and 18 of SR had
precipitating antibodies to Ul-RNP. Twenty-six of 28 positive sera from scleroder-
ma and 13 of 18 positive sera from SR reacted with the 68 kD Ul-RNP associated
protein. Six of 7 sera which did not react with 68 kD protein recognized other
Ul-RNP associated proteins. Three patients (2.5%) with scleroderma had both
anti-Scl-70 and anti-Ul-RNP antibodies. ACA was detected in 17 of scleroderma
and 18 of SR by immunofluorescence. Antibody to 17 kD protein was demon-