DOI Seite / Zitierlink:
https://doi.org/10.11588/diglit.48120#0023
Abstracts
5
52 kDa species in both MOLT-4 and HeLa cell extracts. In addition, rare SS-A/Ro
sera also recognized a weaker antigenic protein of 49 kDa, which was distinct
from the 47 kDa SS-B/La protein. From a A Zap cDNA library derived from
mRNA of human HepG2 cells, a clone Cl was identified by antibody screening
using anti-52 kDa specific anti-SS-A/Ro sera. After several rounds of plaque
purification and using a multispecific SS-A/SS-B autoimmune serum, affinity
purified antibody to the Cl-encoded fusion protein recognized only the 52 kDa
SS-A/Ro protein of MOLT-4 cells in immunoblotting. Bluescript plasmid pCl
rescued from the A Zap phage contained a cDNA insert of 1.5 kb and its restric-
tion map was different from those of the known cDNA clones of a 60 kDa = SS-
A/Ro protein and SS-B/La. Hybridization studies also showed that Cl cDNA in-
sert was not related to the cDNA of 60 kDa SS-A/Ro and SS-B/La. Expression
of pCl plasmid in E. Coli showed that it encoded for a 46 kDa truncated ^-galac-
tosidase fusion protein and, therefore, could represent about 80% of the 52 kDa
SS-A/Ro protein. When partially purified Cl fusion protein was used as antigen
substrate for immunoblotting, 7 anti-SS-A/Ro sera with anti-52 kDa specificity
reacted with the 46 kDa fusion protein and its degradation products, 3 anti-
SS-A/Ro sera with no anti-52 kDa specificity and other negative serum controls
did not react. Southern blot analysis of human peripheral blood DNA restricted
with EcoRI or Hindlll enzymes suggested there are probably only 1 or 2 genes
for the 52 kDA protein. Further analysis in the components of SS-A/Ro antigens
may lead to an understanding of their cellular function and their relationship with
the autoimmune response in these rheumatic diseases.
Autoantibodies to Nuclear Envelope Proteins: Characterization and
Clinical Significance
J. C. Courvalin1, M.-N. Guilly2, F. Danon3, Ch. Andre4, J.-C. Brouet3
and K. Lassoued3
‘CNRS, Gif-sur-Yvette, France and Rockefeller University, New York, NY, USA
2 CEA, Fontenay-aux-Roses, France
3Hopital Saint-Louis, Paris, France
4Hopital Henri Mondor, Paris, France
Significant progress has been made in understanding both the function of
specific nuclear envelope components and their biochemical composition.
Macromolecular traffic into and out of the nucleus is now known to be regulated
5
52 kDa species in both MOLT-4 and HeLa cell extracts. In addition, rare SS-A/Ro
sera also recognized a weaker antigenic protein of 49 kDa, which was distinct
from the 47 kDa SS-B/La protein. From a A Zap cDNA library derived from
mRNA of human HepG2 cells, a clone Cl was identified by antibody screening
using anti-52 kDa specific anti-SS-A/Ro sera. After several rounds of plaque
purification and using a multispecific SS-A/SS-B autoimmune serum, affinity
purified antibody to the Cl-encoded fusion protein recognized only the 52 kDa
SS-A/Ro protein of MOLT-4 cells in immunoblotting. Bluescript plasmid pCl
rescued from the A Zap phage contained a cDNA insert of 1.5 kb and its restric-
tion map was different from those of the known cDNA clones of a 60 kDa = SS-
A/Ro protein and SS-B/La. Hybridization studies also showed that Cl cDNA in-
sert was not related to the cDNA of 60 kDa SS-A/Ro and SS-B/La. Expression
of pCl plasmid in E. Coli showed that it encoded for a 46 kDa truncated ^-galac-
tosidase fusion protein and, therefore, could represent about 80% of the 52 kDa
SS-A/Ro protein. When partially purified Cl fusion protein was used as antigen
substrate for immunoblotting, 7 anti-SS-A/Ro sera with anti-52 kDa specificity
reacted with the 46 kDa fusion protein and its degradation products, 3 anti-
SS-A/Ro sera with no anti-52 kDa specificity and other negative serum controls
did not react. Southern blot analysis of human peripheral blood DNA restricted
with EcoRI or Hindlll enzymes suggested there are probably only 1 or 2 genes
for the 52 kDA protein. Further analysis in the components of SS-A/Ro antigens
may lead to an understanding of their cellular function and their relationship with
the autoimmune response in these rheumatic diseases.
Autoantibodies to Nuclear Envelope Proteins: Characterization and
Clinical Significance
J. C. Courvalin1, M.-N. Guilly2, F. Danon3, Ch. Andre4, J.-C. Brouet3
and K. Lassoued3
‘CNRS, Gif-sur-Yvette, France and Rockefeller University, New York, NY, USA
2 CEA, Fontenay-aux-Roses, France
3Hopital Saint-Louis, Paris, France
4Hopital Henri Mondor, Paris, France
Significant progress has been made in understanding both the function of
specific nuclear envelope components and their biochemical composition.
Macromolecular traffic into and out of the nucleus is now known to be regulated