TY - JOUR
T1 - Naturally occurring masked antibodies in murine sera recognize a component of the mitotic spindle apparatus
AU - Munns, Theodore W.
AU - Liszewski, M. Kathryn
AU - Freeman, Sandra K.
N1 - Funding Information:
ACKNOWLEDGEMENTS The authors gratefully Jeffrey Kaine for their advice and critical investigations. This research was supported the National Institutes of Health and by tne Arthritis Research.
PY - 1986/4/14
Y1 - 1986/4/14
N2 - Antibody activities previously masked in autoimmune MRL and normal Balb c mice were expressed by briefly subjecting their sera to acidic (pH 2.0) or alkaline (pH 12.0) environments. An enzyme-linked immunosorbent assay (ELISA) revealed that these pH-expressable immunoglobulins reacted with specific nucleotide-BSA antigens (primarily 5′-AMP, -GMP, -TMP) but not with single (ss)- or double (ds)-stranded DNAs or with unconjugated BSA. ELISA analysis of pH-expressed antibodies purified via GMP-BSA/Sepharose indicated that they bound not only to the homologous hapten (GMP) but to AMP and TMP as well, i.e., anti-pAGT antibodies. Further, indirect immunofluorescent assays (IIF) with fixed HEp-2 cells demonstrated that purified anti-pAGT antibodies recognized an epitope within the mitotic spindle apparatus. These results document the existence of a previously undefined masked antibody population in murine sera with specificities directed toward certain nucleotides and a component of the mitotic spindle apparatus. Last, these anti-pAGT (or anti-mitotic spindle) antibodies are not restricted to murine systems inasmuch as they have been detected in every human serum (>100 samples) examined thus far.
AB - Antibody activities previously masked in autoimmune MRL and normal Balb c mice were expressed by briefly subjecting their sera to acidic (pH 2.0) or alkaline (pH 12.0) environments. An enzyme-linked immunosorbent assay (ELISA) revealed that these pH-expressable immunoglobulins reacted with specific nucleotide-BSA antigens (primarily 5′-AMP, -GMP, -TMP) but not with single (ss)- or double (ds)-stranded DNAs or with unconjugated BSA. ELISA analysis of pH-expressed antibodies purified via GMP-BSA/Sepharose indicated that they bound not only to the homologous hapten (GMP) but to AMP and TMP as well, i.e., anti-pAGT antibodies. Further, indirect immunofluorescent assays (IIF) with fixed HEp-2 cells demonstrated that purified anti-pAGT antibodies recognized an epitope within the mitotic spindle apparatus. These results document the existence of a previously undefined masked antibody population in murine sera with specificities directed toward certain nucleotides and a component of the mitotic spindle apparatus. Last, these anti-pAGT (or anti-mitotic spindle) antibodies are not restricted to murine systems inasmuch as they have been detected in every human serum (>100 samples) examined thus far.
UR - http://www.scopus.com/inward/record.url?scp=0022456646&partnerID=8YFLogxK
U2 - 10.1016/0006-291X(86)90888-0
DO - 10.1016/0006-291X(86)90888-0
M3 - Article
C2 - 3518712
AN - SCOPUS:0022456646
SN - 0006-291X
VL - 136
SP - 145
EP - 150
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -