Carbohydrate-mediated recognition of a circulating placental alkaline phosphatase-immunoglobulin M complex.
We detected an abnormal alkaline phosphatase (AP) electrophoretically in the serum of a patient with rheumatoid arthritis, who had a macromolecular AP linked with immunoglobulin M (IgM) bearing a kappa light chain. The IgM isolated from the AP-IgM complex in the patient's serum reacted apparently with all of the AP isozymes tested, i.e. those originating in the liver, bone, intestine and placenta, but the alpha-mannosidase-treated IgM from the patient's serum bound to placental AP (PAP) alone. This suggests that untreated IgM recognizes multivalent epitopes of the AP and that the complex of AP with alpha-mannosidase-treated IgM is a specific antibody-antigen complex. In order to investigate further the multivalent binding capacity for the PAP-untreated IgM complex, we prepared a monoclonal antibody (MoAb) against PAP and identified it as an IgM with a kappa light chain. The binding affinities and their circulating half-lives of the synthetic complexes of PAP and respective MoAbs were examined with and without treatment with several glycosidases. The untreated MoAb bearing IgM had binding affinity for all of the AP isozymes tested, while alpha-mannosidase-treated IgM attached only to PAP, the same as the IgM isolated from the PAP-IgM complex in the patient's serum. The circulating clearance of the PAP-IgM complex in rabbits was faster than either component alone. In addition, the PAP-IgM complex treated with alpha-mannosidase was found to have the shortest half-life of all the complexes of PAP and Igs treated with the several glycosidases tested. These results suggest that the formation of the PAP-IgM complex as an enzyme-linked antibody and the clearance of the complex in vivo are dependent on the sugar moieties of the Igs.
Koyama
I
I
Department of Medical Technology, Junior College, Saitama, Japan.
Sato
M
M
Sakai
H
H
Nagata
A
A
Miura
M
M
Sakagishi
Y
Y
Komoda
T
T
D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis*; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis* / Q000378:metabolism; D001172:Arthritis, Rheumatoid / Q000201:enzymology / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005260:Female; D006026:Glycoside Hydrolases / Q000737:chemistry; D006207:Half-Life; D006801:Humans; D007075:Immunoglobulin M / Q000032:analysis*; D007145:Immunoglobulin kappa-Chains / Q000276:immunology*; D007527:Isoenzymes / Q000032:analysis*; D010920:Placenta / Q000201:enzymology*; D011233:Precipitin Tests; D011247:Pregnancy; D011248:Pregnancy Complications / Q000201:enzymology / Q000276:immunology
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000939:Epitopes; D007075:Immunoglobulin M; D007145:Immunoglobulin kappa-Chains; D007527:Isoenzymes; D000469:Alkaline Phosphatase; D006026:Glycoside Hydrolases