Anti-prolactin (PRL) autoantibody-binding sites (epitopes) on PRL molecule in macroprolactinemia.
Macroprolactinemia, in which serum prolactin (PRL) mainly consists of PRL with a molecular mass greater than 100 kDa, has been demonstrated to be associated with hyperprolactinemia. We previously reported that anti-PRL autoantibody is the major cause of macroprolactinemia. In this study, the autoantibody-binding sites (epitopes) on the PRL molecule were examined using deletion mutant PRL. The sera from 159 patients with hyperprolactinemia were screened for macroprolactinemia using the polyethylene glycol method and 18 patients (11%) were diagnosed with macroprolactinemia. The sera from these patients were incubated with glutathione S-transferase-human prolactin (hPRL) fragment fusion proteins immobilized on glutathione sepharose and the amounts of bound immunoglobulin G (IgG) were measured using ELISA. IgG was bound to full-length hPRL1-199 in significantly greater amounts in sera from 14 of 18 patients with macroprolactinemia than in controls. hPRL, but not PRL of other species such as bovine, porcine, rat, or human GH, dose-dependently displaced the binding, suggesting that these patients had hPRL-specific autoantibodies. Deletion of 34 amino acid residues from N-and/or C-terminals significantly reduced the binding and N- or C-terminal fragment alone showed partial but significant binding, suggesting that the major epitopes recognized by anti-PRL autoantibodies are located in both N- and C-terminal residues of the PRL molecule.
Hattori
N
N
Department of Pharmacology, Kansai Medical University, 10-15 Fumizono-cho, Moriguchi-City, Osaka 570-8506, Japan. hattorin@takii.kmu.ac.jp
Nakayama
Y
Y
Kitagawa
K
K
Ishihara
T
T
Saiki
Y
Y
Inagaki
C
C
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D001323:Autoantibodies / Q000378:metabolism*; D001682:Biological Availability; D016022:Case-Control Studies; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000032:analysis*; D005260:Female; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D006966:Hyperprolactinemia / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D011388:Prolactin / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D011981:Receptors, Prolactin / Q000032:analysis / Q000378:metabolism; D011994:Recombinant Proteins
D001323:Autoantibodies; D017931:DNA Primers; D000939:Epitopes; D011981:Receptors, Prolactin; D011994:Recombinant Proteins; C060678:prolactin, polymeric; D011388:Prolactin; D005982:Glutathione Transferase