A bioactive 60-kilodalton prolactin species is preferentially secreted in cultures of mitogen-stimulated and nonstimulated peripheral blood mononuclear cells from subjects with systemic lupus erythematosus.
We have evaluated the production of PRL by human peripheral mononuclear cells (PBMNC) from normal subjects and patients with systemic lupus erythematosus (SLE). Conditioned medium prepared from basal and Con-A-stimulated PBMNC was assessed for the presence of PRL-like by its ability to stimulate growth of PRL-responsive Nb2 rat lymphoma cells. In the presence or absence of Con-A, SLE PBMNC secrete significantly higher (P < 0.001) amounts of bioactive PRL-like species than normal cells. Growth of Nb2 cells by conditioned medium was inhibited with specific antiserum to human PRL. Western blotting using a polyclonal antibody to human PRL revealed a single 60-kDa PRL-like species in both normal and SLE PBMNC extracts, the immunoreactivity of which was preferentially found in SLE subjects. With the use of reverse transcription-PCR an expected 633-bp band was observed, and its similarity to pituitary PRL was further confirmed by Southern blot analysis with human PRL complementary DNA as a probe. We conclude that a high molecular mass PRL-like species is synthesized and secreted by PBMNC, and patients with SLE have an increased secretion of lymphocyte-derived PRL-like material.
Larrea
F
F
Department of Reproductive Biology, Instituto Nacional de la Nutrición Salvador Zubirán, Mexico City, Mexico.
Martínez-Castillo
A
A
Cabrera
V
V
Alcocer-Varela
J
J
Queipo
G
G
Cariño
C
C
Alarcón-Segovia
D
D
D000293:Adolescent; D000328:Adult; D000818:Animals; D001681:Biological Assay; D015139:Blotting, Southern; D015153:Blotting, Western; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology; D017077:Culture Media, Conditioned; D005260:Female; D015870:Gene Expression; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008180:Lupus Erythematosus, Systemic / Q000097:blood*; D008223:Lymphoma / Q000473:pathology; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D011388:Prolactin / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats
D017077:Culture Media, Conditioned; D012333:RNA, Messenger; D003208:Concanavalin A; D011388:Prolactin