Herrera M F, Grant C S, Maercklein P, van Heerden J A, Fitzpatrick L A
Department of Surgery, Mayo Clinic, Rochester, Minn. 55905.
Surgery. 1993 Apr;113(4):410-8.
We have recently validated a reverse hemolytic plaque assay (RHPA) for the quantitative measurement of parathyroid hormone release from single isolated parathyroid cells. The information regarding secretory behavior of normal and abnormal parathyroid tissue has been based previously on fluorescent spectroscopy or quantification of immunoreactive parathyroid hormone by radioimmunoassay. Thus, individual differences in secretion and assessment of viability are not clearly measured in these types of experiments. The RHPA has the advantage of allowing measurements of secretory activity in individual viable parathyroid cells. In this report, we have modified the RHPA to measure the release of parathyroid hormone from human cells. This is the first instance of measurement of hormone secretion from human cells, and we present data indicating the specificity of the assay for human parathyroid hormone. The response of normal human parathyroid cells to alterations in extracellular calcium was assessed by two parameters: the ability of an individual cell to secrete parathyroid hormone in response to a stimulus and the recruitment of normal human cells to release parathyroid hormone. This assay will allow elucidation of secretory dynamics in normal and abnormal glands, which may help us to understand the mechanisms involved in parathyroid dysfunction.
我们最近验证了一种反向溶血空斑试验（RHPA），用于定量测量单个分离甲状旁腺细胞释放的甲状旁腺激素。以前，关于正常和异常甲状旁腺组织分泌行为的信息是基于荧光光谱法或通过放射免疫测定法对免疫反应性甲状旁腺激素进行定量。因此，在这些类型的实验中，分泌的个体差异和活力评估并未得到明确测量。RHPA的优点是能够测量单个存活甲状旁腺细胞的分泌活性。在本报告中，我们对RHPA进行了改进，以测量人细胞中甲状旁腺激素的释放。这是首次测量人细胞激素分泌的实例，我们提供的数据表明了该测定法对人甲状旁腺激素的特异性。通过两个参数评估正常人甲状旁腺细胞对细胞外钙变化的反应：单个细胞响应刺激分泌甲状旁腺激素的能力以及募集正常人细胞释放甲状旁腺激素的情况。该测定法将有助于阐明正常和异常腺体中的分泌动态，这可能有助于我们了解甲状旁腺功能障碍所涉及的机制。
