Growth hormone augments superoxide anion secretion of human neutrophils by binding to the prolactin receptor.
Recombinant human growth hormone (HuGH) and human prolactin (HuPRL), but not GH of bovine or porcine origin, prime human neutrophils for enhanced superoxide anion (O2-) secretion. Since HuGH, but not GH of other species, effectively binds to the HuPRL receptor (HuPRL-R), we used a group of HuGH variants created by site-directed mutagenesis to identify the receptor on human neutrophils responsible for HuGH priming. A monoclonal antibody (MAb) directed against the HuPRL-R completely abrogated O2- secretion by neutrophils incubated with either HuGH or HuPRL, whereas a MAb to the HuGH-R had no effect. The HuGH variant K172A/F176A, which has reduced affinity for both the HuGH-binding protein (BP) and the HuPRL-BP, was unable to prime human neutrophils. This indicates that priming is initiated by a ligand-receptor interaction, the affinity of which is near that defined for receptors for PRL and GH. Another HuGH variant, K168A/E174A, which has relatively low affinity for the HuPRL-BP but slightly increased affinity for the HuGH-BP, had much reduced ability to prime neutrophils. In contrast, HuGH variant E56D/R64M, which has a similar affinity as wild-type HuGH for the HuPRL-BP but a lower affinity for the HuGH-BP, primed neutrophils as effectively as the wild-type HuGH. Finally, binding of HuGH to the HuPRL-BP but not to the HuGH-BP has been shown to be zinc dependent, and priming of neutrophils by HuGH was also responsive to zinc. Collectively, these data directly couple the binding of HuGH to the HuPRL-R with one aspect of functional activation of human target cells.
Fu
Y K
YK
Department of Animal Sciences, University of Illinois, Urbana 61801.
Arkins
S
S
Fuh
G
G
Cunningham
B C
BC
Wells
J A
JA
Fong
S
S
Cronin
M J
MJ
Dantzer
R
R
Kelley
K W
KW
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured; D013006:Growth Hormone / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D008223:Lymphoma / Q000473:pathology; D009504:Neutrophils / Q000378:metabolism*; D011981:Receptors, Prolactin / Q000502:physiology*; D013481:Superoxides / Q000378:metabolism*; D013552:Swine; D015032:Zinc / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D011981:Receptors, Prolactin; D013481:Superoxides; D013006:Growth Hormone; D015032:Zinc