Wilson G B, Fudenberg H H, Parise M T, Floyd E
J Clin Invest. 1981 Jul;68(1):171-83. doi: 10.1172/jci110233.
Cultured mononuclear cells (MNC) from individuals homozygous or heterozygous for the defective gene causing the inherited disease cystic fibrosis (CF) synthesize three unusual "mediators" termed ciliary dyskinesia substances (CDS), which markedly affect tracheal mucociliary systems in vitro. MNC cultures from normal healthy controls do not accumulate any CDS, whereas MNC cultures from non-CF patients controls with pulmonary disease synthesized at least one CDS. The possible involvement of the CDS in pulmonary disease is being investigated. In this study, we sought to determine whether the CDS could be chemoattractants for polymorphonuclear neutrophils (PMN), since they have characteristics in common with known chemoattractants generated by alveolar macrophages. Our analyses of crude MNC culture supernates indicated that cultures from both CF genotypes accumulate significantly higher levels of PMN chemoattractants than do analogous cultures from normal healthy controls. CF homozygote MNC also generated more activity than MNC from patient controls with chronic pulmonary disease. Fractionation of MNC culture supernates by gel permeation chromatography and characterization of active fractions demonstrated six distinct PMN chemoattractants in cultures from CF genotypes; five were also present in patient control and four in normal healthy control cultures. The excessive chemoattractant activity in MNC cultures from CF genotypes and patient controls was due to several different substances produced by monocytes: (a) two components of 1,000-3,500 mol wt. (b) two fragments of C5, and (c) a fragment of C3. One C5 fragment had ciliary dyskinesia activity, the other did not. The C3 fragment chemoattractant also had ciliary dyskinesia activity and was not found in MNC cultures from patient controls. A third CDS, Which is CF-specific (5,000 mol wt), was neither chemotactic not chemokinetic and did not inhibit random PMN migration; however, fractions containing this CF-specific CDS completely inhibited PMN chemotaxis in response to three different chemoattractants. We conclude that all of the CDS can potentially play a role in the pathophysiology of lung disease, as judged by their effects on PMN movement in vitro.
从患有遗传性疾病囊性纤维化（CF）的缺陷基因纯合子或杂合子个体中培养的单核细胞（MNC）可合成三种不同寻常的“介质”，称为纤毛运动障碍物质（CDS），它们在体外能显著影响气管黏液纤毛系统。来自正常健康对照者的MNC培养物不会积累任何CDS，而来自患有肺部疾病的非CF患者对照的MNC培养物则至少合成一种CDS。目前正在研究CDS在肺部疾病中的可能作用。在本研究中，我们试图确定CDS是否可能是多形核中性粒细胞（PMN）的趋化因子，因为它们具有与肺泡巨噬细胞产生的已知趋化因子相同的特征。我们对粗制MNC培养上清液的分析表明，来自两种CF基因型的培养物积累的PMN趋化因子水平明显高于来自正常健康对照的类似培养物。CF纯合子MNC产生的活性也高于患有慢性肺部疾病的患者对照的MNC。通过凝胶渗透色谱法对MNC培养上清液进行分级分离并对活性级分进行表征，结果表明来自CF基因型的培养物中有六种不同的PMN趋化因子；患者对照培养物中也存在五种，正常健康对照培养物中存在四种。CF基因型和患者对照的MNC培养物中趋化因子活性过高是由于单核细胞产生的几种不同物质所致：（a）两种分子量为1000 - 3500的成分；（b）C5的两个片段；（c）C3的一个片段。一个C5片段具有纤毛运动障碍活性，另一个则没有。C3片段趋化因子也具有纤毛运动障碍活性，且在患者对照的MNC培养物中未发现。第三种CDS是CF特异性的（分子量为5000），既没有趋化作用也没有促动作用，并且不抑制PMN的随机迁移；然而，含有这种CF特异性CDS的级分完全抑制了PMN对三种不同趋化因子的趋化作用。我们得出结论，从它们对体外PMN运动的影响来看，所有的CDS都可能在肺部疾病的病理生理学中发挥作用。
