Vitamin D receptor expression in human lymphocytes. Signal requirements and characterization by western blots and DNA sequencing.
The signals controlling the expression of the receptor protein for 1 alpha,25-dihydroxyvitamin D3 in normal human lymphocytes and the relationship of this protein to the classical vitamin D receptor were examined. Lymphocytes activated with the OKT3 antibody to the T-cell antigen receptor expressed fewer binding sites as compared to lymphocytes that were activated by the polyclonal activator phytohemagglutinin (PHA). However, combination of OKT3 and phorbol myristate acetate produced a concentration of binding sites similar to the PHA-activated cells. The receptor from OKT3 and OKT3 + phorbol myristate acetate-activated lymphocytes exhibited decreased binding to DNA-cellulose compared to PHA-activated lymphocytes. In lymphocytes activated either by PHA or OKT3 (but not in resting cells), a 50-kDa species cross-reacting with a monoclonal antibody against the intestinal vitamin D receptor was detected. Finally, RNA from activated lymphocytes was amplified by polymerase chain reaction using oligonucleotide primers flanking the 196 base pair long region encoding the DNA-binding domain of the human intestinal receptor. The amplified product showed an identical nucleotide sequence to the DNA-binding domain of the human intestinal receptor. These findings suggest that expression of the 1,25-(OH)2D3 receptor in lymphocytes is triggered by distinct and contingent signals, and that the protein and the mRNA encoding it are identical to the classical vitamin D receptor.
Yu
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Section of Endocrinology and Metabolism, Veterans Administration Medical Center, Indianapolis, Indiana 46202.
Mocharla
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Hustmyer
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Manolagas
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SC
D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015153:Blotting, Western; D002117:Calcitriol / Q000378:metabolism*; D002853:Chromatography, Liquid; D004247:DNA / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D010835:Phytohemagglutinins / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid / Q000235:genetics* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D010835:Phytohemagglutinins; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid; D004247:DNA; D002117:Calcitriol; D013755:Tetradecanoylphorbol Acetate