Immuno-enhancement and -inhibition of GH-releasing factor by site-directed anti peptide antibodies in vivo and in vitro.
It is now well established that specific antibodies and binding proteins can potentiate rather than inhibit hormone activity. In order to investigate this phenomenon further, the current study was undertaken using a hormone with a characterised structure, in terms of receptor binding, and for which activity has already been manipulated in specific ways (prolongation of half-life, increased receptor affinity) using synthetic hormone analogues. GH-releasing factor (GRF) is a 40 or 44 residue peptide and is, together with somatostatin, responsible for the regulation of GH secretion. The effects of site-directed anti peptide antibodies were determined on the activity of GRF in vivo and in vitro as GH release. The peptide regions of GRF were: 1-14 (part of putative receptor-binding region) and 31-44 and 35-44 (sites thought to be distant from the receptor-binding region). Five sheep were administered GRF (1 microgram/kg), anti peptide immunoglobulin (Ig; a calculated tenfold excess binding to GRF dose), or GRF together with anti peptide Ig (preincubated for 1 h). GRF induced a significant increase in plasma GH concentration over the next 240 min, this was abolished when GRF was administered with anti 1-14 Ig (P < 0.05) and augmented (P < 0.05) when GRF was administered with anti 35-44 Ig; anti 31-44 had no effect on GRF activity. Anti 35-44 Ig alone induced an increase in GH secretion which was equivalent to that for GRF alone, implying that the antibody had interacted and potentiated with endogenous GRF. The Ig effects on exogenous GRF activity were confirmed for GH release in vitro using primary cultures of sheep pituitary cells, except that anti 31-44 Ig also augmented GH release (P < 0.05) when co-administered with GRF. (ABSTRACT TRUNCATED AT 250 WORDS)
Pell
J M
JM
Department of Cellular Physiology, Babraham Institute, Cambridge, UK.
James
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002478:Cells, Cultured; D005260:Female; D013006:Growth Hormone / Q000378:metabolism*; D013007:Growth Hormone-Releasing Hormone / Q000276:immunology / Q000378:metabolism*; D006728:Hormones / Q000276:immunology; D010446:Peptide Fragments; D010902:Pituitary Gland / Q000378:metabolism; D011863:Radioimmunoassay; D012756:Sheep / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D006728:Hormones; D010446:Peptide Fragments; D013006:Growth Hormone; D013007:Growth Hormone-Releasing Hormone