Studies on the Regulation of Immune Functions by Growth Hormone.

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Growth hormone-dependent proliferation of IM-9 cells under low-serum conditions was inhibited by the addition of phorbol diesters. The phorbol diesters were found to down-regulate the growthhormone receptor levels. These regulations by phorbol diesters were inhibited by staurosporine or sphingosine, suggesting the involvement of C-kinases. It was also found that phosphorylation of a 55 kDa protein, which was associated with HGHR, was increased.Monoclonal anti-peptide antibodies against human growth hormone receptor(HGHR)were prepared and characterized. Among them, the monoclonal antibody GHRP2-88 was reactive with the extracellular domain of HGHR and the most efficient when used for immunoblotting of IM-9 cell membrane proteins.Monoclonal antibodies against ubiquitin were prepared for the purpose of elucidating the role of hGHR-bound ubiquitin.