DOI Page / Citation link:
https://doi.org/10.11588/diglit.48120#0052
34
Molecular and Cell Biology of Autoantibodies and Autoimmunity
that nucleic acid antigens were not detached from a solid phase by washing with
0.5 M NaCl solution indicated that the reductions of binding of anti-DNA an-
tibodies in both MoAbs and naturally-occurring antibodies were really due to
dissociation of the antibodies from immune complexes. This is the first demon-
stration that DNA epitopes recognized by naturally-occurring antibodies in both
systemic lupus erythematosus (SLE) and its model mice are sensitive to neutral
NaCl concentrations. This novel trait of naturally-occurring antibodies will be
very useful in studies on the nature of immune complexes in the sera and kidney
of cases of SLE.
Immunoblotting Analysis in Antigenic Components of Circulating
Immune Complexes of Rheumatoid Arthritis and Systemic Lupus
Erythematosus Patients
R. Kasukawa, Y. Sato, Y. Okubo, and T. Nishimaki
Second Department of Internal Medicine, Fukushima Medical College, Fukushima, Japan
In order to analyze the antigenic components in circulating immune com-
plexes (IC) in patients with rheumatoid arthritis (RA) and systemic lupus
erythematosus (SLE), immune complexes in serum or joint fluid (JF) were
isolated in using solid phase Clq tubes and analyzed for their components
through immunoblotting method.
In separation of RA IC on SPS-PAGE, components of 73, 54 and 27 kD
bands were identified as // chain of IgM, y chain of IgG, and L chain of Ig respec-
tively. These components were transfered to cellulose acetate membrane and
allowed to react with IgM fraction of RA serum or RA JF obtained through
Sephadex G-200 filtration under acid condition. The IgM fraction could react
with 54 kD (y chain of IgG) component but not with other components.
Five SLE IC samples taken from 5 patients who have anti-DNA antibody
commonly besides other antibodies were tested on the cellulose acetate membrane
against HeLa extract separated on SDS-PAGE. Several bands were demonstrated
on the membrane by the anti-human IgG antibody with mobilities ranging from
76 to 16kD. However, in the reaction between the purified plasmid DNA
separated on SDS-PAGE and SLE IC in serum, only 16kD band was identified
by the anti-human IgG antibody.
The DNA in SLE IC was labelled by 32P through multiprime labelling meth-
od and separated on SDS-PAGE. The autoradiography showed wide bands with
Molecular and Cell Biology of Autoantibodies and Autoimmunity
that nucleic acid antigens were not detached from a solid phase by washing with
0.5 M NaCl solution indicated that the reductions of binding of anti-DNA an-
tibodies in both MoAbs and naturally-occurring antibodies were really due to
dissociation of the antibodies from immune complexes. This is the first demon-
stration that DNA epitopes recognized by naturally-occurring antibodies in both
systemic lupus erythematosus (SLE) and its model mice are sensitive to neutral
NaCl concentrations. This novel trait of naturally-occurring antibodies will be
very useful in studies on the nature of immune complexes in the sera and kidney
of cases of SLE.
Immunoblotting Analysis in Antigenic Components of Circulating
Immune Complexes of Rheumatoid Arthritis and Systemic Lupus
Erythematosus Patients
R. Kasukawa, Y. Sato, Y. Okubo, and T. Nishimaki
Second Department of Internal Medicine, Fukushima Medical College, Fukushima, Japan
In order to analyze the antigenic components in circulating immune com-
plexes (IC) in patients with rheumatoid arthritis (RA) and systemic lupus
erythematosus (SLE), immune complexes in serum or joint fluid (JF) were
isolated in using solid phase Clq tubes and analyzed for their components
through immunoblotting method.
In separation of RA IC on SPS-PAGE, components of 73, 54 and 27 kD
bands were identified as // chain of IgM, y chain of IgG, and L chain of Ig respec-
tively. These components were transfered to cellulose acetate membrane and
allowed to react with IgM fraction of RA serum or RA JF obtained through
Sephadex G-200 filtration under acid condition. The IgM fraction could react
with 54 kD (y chain of IgG) component but not with other components.
Five SLE IC samples taken from 5 patients who have anti-DNA antibody
commonly besides other antibodies were tested on the cellulose acetate membrane
against HeLa extract separated on SDS-PAGE. Several bands were demonstrated
on the membrane by the anti-human IgG antibody with mobilities ranging from
76 to 16kD. However, in the reaction between the purified plasmid DNA
separated on SDS-PAGE and SLE IC in serum, only 16kD band was identified
by the anti-human IgG antibody.
The DNA in SLE IC was labelled by 32P through multiprime labelling meth-
od and separated on SDS-PAGE. The autoradiography showed wide bands with