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囊性纤维化:牡蛎纤毛试验研究

Cystic fibrosis: studies with the oyster ciliary assay.

作者信息

Bowman B H, Lankford B J, McNeely M C, Carson S D, Barnett D R, Berg K

出版信息

Clin Genet. 1977 Dec;12(6):333-43. doi: 10.1111/j.1399-0004.1977.tb00952.x.

Abstract

Bioassays using ciliary systems have detected a factor or factors in cystic fibrosis (CF) sera and tissue culture medium derived from CF cells. The typical shortcomings of an assay measuring biological activity have been studied, and the means to overcome the weaknesses of the oyster gill cilia assay have been established. The presence of the cystic fibrosis mucociliary inhibitor (CFMI) in experimental fractions may be determined by accepting data from only those assays in which authentic CF and normal (non-CF) fractions give defined reactions, by measuring the reaction of each sample at least three times, and by examining each experimental sample at a protein concentration greater than the minimum established in this study. The relative concentrations of the CFMI present in the first steps of purification of serum and medium have been calculated in terms of units of inhibition. Generally, the units of inhibition present in serum and medium fractions from heterozygotes are close to one-half of that in fractions from homozygous sources. Analogous fractions concentrated from a normal (non-CF) source never inhibited mucociliary activity, even when tested at nearly 100 times the CF concentration. Ciliary assays utilizing oyster gills are essential for monitoring fractionation procedures aimed at purifying the CFMI, and have been shown to be capable and reliable enough to do so.

摘要

使用纤毛系统的生物测定法已在囊性纤维化(CF)血清和源自CF细胞的组织培养基中检测到一种或多种因子。已研究了测量生物活性的测定法的典型缺点,并已建立克服牡蛎鳃纤毛测定法弱点的方法。实验组分中囊性纤维化黏液纤毛抑制剂(CFMI)的存在可通过仅接受来自那些真实CF和正常(非CF)组分给出明确反应的测定法的数据、通过至少测量每个样品的反应三次以及通过在高于本研究确定的最低蛋白质浓度下检查每个实验样品来确定。已根据抑制单位计算了血清和培养基纯化第一步中存在的CFMI的相对浓度。一般来说,杂合子血清和培养基组分中的抑制单位接近纯合子来源组分中的抑制单位的一半。即使在接近CF浓度100倍的情况下进行测试,从正常(非CF)来源浓缩的类似组分也从未抑制黏液纤毛活性。利用牡蛎鳃的纤毛测定法对于监测旨在纯化CFMI的分级分离程序至关重要,并且已证明其有足够的能力和可靠性来做到这一点。

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