The impact of FCN2 polymorphisms and haplotypes on the ficolin-2 serum levels
The impact of FCN2 polymorphisms and haplotypes on the ficolin-2 serum levels
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DOI：
10.1111/j.1365-3083.2007.01915.x
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发表时间：
2007-04-01
影响因子：
3.7
通讯作者：
Garred, P.
中科院分区：
文献类型：
--
作者：
Munthe-Fog, L.;Hummelshoj, T.;Garred, P.
Ficolin-2 （L-ficolin）来源于FCN2基因，是一种在人血清中发现的先天性免疫模式识别分子，其血清中的个体间差异似乎受遗传控制。为了验证和扩展这一发现，我们开发了一种用于检测血清样本中人Ficolin-2的夹心ELISA，并通过Taq man小槽结合物试验和测序鉴定了FCN2基因型。血清样品应用凝胶渗透层析，用ELISA、SDS-PAGE和随后的Western blotting分析组分。在214名丹麦献血者中，测定了Ficolin-2的中位血清浓度为5.4 μ g/ml（范围：1.0-12.2 μ g/ml）。ELISA、SDS-PAGE和凝胶渗透层析组分的Western blot分析表明，Ficolin-2由共价和非共价连接的Ficolin-2低聚物组成，独立于单个基因型。血清浓度的变化与FCN2基因启动子的三个多态性和FCN2基因结构部分的一个多态性有关。进一步分析表明，同一单倍型上的两个特定等位基因决定了低的Ficolin-2浓度。我们的研究结果表明，Ficolin-2浓度的个体间变异与FCN2基因启动子和结构部分的多态性有关。
Ficolin-2 (L-ficolin), derived from the FCN2 gene, is an innate immunity pattern recognition molecule found in human serum in which inter-individual variation in serum appears to be under genetic control. To validate and extend this finding, we developed a sandwich ELISA for detection of human Ficolin-2 in serum samples and identified FCN2 genotypes with a Taq Man-based minor groove binder assay and by sequencing. Serum samples were applied to gel-permeation chromatography and fractions were analysed by an ELISA, SDS-PAGE and subsequently Western blotting. In 214 Danish blood donors, the median Ficolin-2 serum concentration was determined to 5.4 mu g/ml (range: 1.0-12.2 mu g/ml). An ELISA, SDS-PAGE and Western blot analysis of gel-permeation chromatography fractions showed that Ficolin-2 comprises a mixture of covalently and non-covalently linked Ficolin-2 oligomers independent of the individual genotypes. The variation in serum concentration was associated with three polymorphisms in the promoter and one polymorphism in the structural part of the FCN2 gene. Further analysis indicated that two particular alleles on the same haplotype determined a low Ficolin-2 concentration. Our results show that inter-individual variation of Ficolin-2 concentration is associated with polymorphisms in the promoter and the structural part of the FCN2 gene.