Piskurich J F, France J A, Tamer C M, Willmer C A, Kaetzel C S, Kaetzel D M
Institute of Pathology, Case Western Reserve University, Cleveland, OH 44106.
Mol Immunol. 1993 Mar;30(4):413-21. doi: 10.1016/0161-5890(93)90071-i.
Transport of secretory IgA into external fluids is mediated by the polymeric immunoglobulin receptor (pIgR) on the surface of mucosal epithelial cells. We studied the mechanism by which interferon-gamma (IFN-gamma) induces pIgR expression in HT-29.74 cells, a subclone of the HT-29 cell line selected for high concns of pIgR. Here we report the isolation of genomic DNA and cDNA clones encoding human pIgR and development of a sensitive ribonuclease protection assay for pIgR mRNA. This assay was used to determine if induction of pIgR by IFN-gamma is mediated by accumulation of pIgR mRNA. After an initial lag of 12 hr, pIgR mRNA increased seven-fold in response to IFN-gamma, reaching a plateau at 24 hr. Concentrations of pIgR protein also increased seven-fold, but the increase was delayed until 48 hr following stimulation with IFN-gamma. Cycloheximide treatment abolished the IFN-gamma induced increase in pIgR mRNA, indicating that induction of pIgR mRNA by IFN-gamma requires de novo protein synthesis. These results suggest that induction of pIgR expression by IFN-gamma involves an increase in steady-state concns of pIgR mRNA via a protein synthesis dependent mechanism.
分泌型IgA向外部液体的转运是由黏膜上皮细胞表面的聚合免疫球蛋白受体(pIgR)介导的。我们研究了干扰素-γ(IFN-γ)诱导HT-29.74细胞中pIgR表达的机制,HT-29.74细胞是从HT-29细胞系中挑选出来的一个亚克隆,其pIgR浓度较高。在此我们报告了编码人pIgR的基因组DNA和cDNA克隆的分离,以及一种针对pIgR mRNA的灵敏核糖核酸酶保护分析方法的建立。该分析方法用于确定IFN-γ对pIgR的诱导是否由pIgR mRNA的积累介导。在最初12小时的延迟后,pIgR mRNA对IFN-γ的反应增加了7倍,在24小时达到平台期。pIgR蛋白的浓度也增加了7倍,但增加延迟到用IFN-γ刺激后48小时。放线菌酮处理消除了IFN-γ诱导的pIgR mRNA增加,表明IFN-γ诱导pIgR mRNA需要从头合成蛋白质。这些结果表明,IFN-γ诱导pIgR表达涉及通过蛋白质合成依赖机制增加pIgR mRNA的稳态浓度。