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未培养和培养的人外周血单个核细胞中的内啡肽样免疫反应性

Endorphin-like immunoreactivities in uncultured and cultured human peripheral blood mononuclear cells.

作者信息

van Woudenberg A D, Hol E M, Wiegant V M

机构信息

Department of Medical Pharmacology, University of Utrecht, The Netherlands.

出版信息

Life Sci. 1992;50(10):705-14. doi: 10.1016/0024-3205(92)90473-3.

Abstract

It has been reported that cells of the immune system produce and release considerable amounts of pro-opiomelanocortin (POMC) -derived peptides in response to coculture with a variety of stimulatory agents. The present study investigated whether extracts of human peripheral blood mononuclear cells (PBMC) contain immunoreactivity for beta-endorphin (beta E) and related peptides. Using four endorphin RIA systems with different specificities, extracts of freshly isolated PBMC and PBMC cultured in the presence or absence of mitogens or of corticotropin releasing factor (CRF) and vasopressin (VP), were analyzed. With a radioimmunoassay (RIA) system directed to the midportion of beta E, immunoreactivity (MP beta E-IR) was readily detectable, although the concentration was extremely low (ca. 200 pg/10(7) cells). beta E immunoreactivity (beta E-IR) and alpha-endorphin immunoreactivity (alpha E-IR), as determined in C-terminally directed RIA systems, were present in even lower concentrations. gamma-Endorphin immunoreactivity (gamma E-IR) was hardly detectable. Of subsets enriched in T-cells, B-cells or monocytes, the highest concentration of MP beta E-IR was detected in extracts of monocytes. Coculture of PBMC with the mitogen Concanavalin A (Con A) or Phytohaemagglutinin (PHA) increased the amount of MP beta E-IR in extracts of the cells. No increase in alpha E-IR, however, was detected, whereas beta E-IR was only increased in extracts of cells cultured in the presence of Con A. No increase, in any of the immunoreactivities, was observed in extracts of PBMC cultured with bacterial lipopolysaccharide (LPS) or with the combination of CRF and VP, both stimuli that have been reported to induce POMC peptides in cultured PBMC. The present data show that human PBMC contain endorphin-like immunoreactivity, but in very small amounts. The extremely low concentrations and the ineffectiveness of LPS and the combination of CRF and VP to increase the endorphin-like immunoreactivity raise questions about the reported capacity of PBMC to synthesize POMC-derived peptides.

摘要

据报道,免疫系统细胞在与多种刺激剂共培养时会产生并释放大量源自阿片促黑激素原(POMC)的肽。本研究调查了人外周血单核细胞(PBMC)提取物中是否含有β-内啡肽(βE)及相关肽的免疫反应性。使用四种具有不同特异性的内啡肽放射免疫分析(RIA)系统,对新鲜分离的PBMC提取物以及在有丝分裂原、促肾上腺皮质激素释放因子(CRF)或血管加压素(VP)存在或不存在的情况下培养的PBMC提取物进行了分析。使用针对βE中部的放射免疫分析(RIA)系统,虽然浓度极低(约200 pg/10⁷个细胞),但免疫反应性(MP βE-IR)很容易检测到。在C末端导向的RIA系统中测定的βE免疫反应性(βE-IR)和α-内啡肽免疫反应性(αE-IR)浓度更低。γ-内啡肽免疫反应性(γE-IR)几乎检测不到。在富含T细胞、B细胞或单核细胞的亚群中,单核细胞提取物中检测到的MP βE-IR浓度最高。PBMC与有丝分裂原刀豆球蛋白A(Con A)或植物血凝素(PHA)共培养会增加细胞提取物中MP βE-IR的量。然而,未检测到αE-IR增加,而βE-IR仅在Con A存在下培养的细胞提取物中增加。在用细菌脂多糖(LPS)或CRF与VP组合培养的PBMC提取物中,未观察到任何免疫反应性增加,这两种刺激剂据报道可在培养的PBMC中诱导POMC肽。目前的数据表明,人PBMC含有内啡肽样免疫反应性,但含量极少。极低的浓度以及LPS和CRF与VP组合增加内啡肽样免疫反应性的无效性,对报道的PBMC合成POMC衍生肽的能力提出了疑问。

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