Structural decomposition and heterogeneity of commercial lipoteichoic acid preparations
Structural decomposition and heterogeneity of commercial lipoteichoic acid preparations
复制标题
DOI：
10.1128/iai.70.2.938-944.2002
复制
发表时间：
2002-02-01
影响因子：
3.1
通讯作者：
Hartung, T
中科院分区：
文献类型：
--
作者：
Morath, S;Geyer, A;Hartung, T
利用疏水相互作用色谱（HIC）和核磁共振波谱对商业制备的脂壁酸（LTA）进行分离，发现LTA的组成和分解结构非常不均匀：化脓性链球菌的LTA平均含量为61%，枯草芽孢杆菌为16%，金黄色葡萄球菌为75%。分解的特点是甘油磷酸单位以及丙氨酸和n -乙酰氨基葡萄糖取代基的损失。所有制剂都不同程度地含有非lta、非脂多糖（LPS）免疫刺激成分，这可以从它们在HIC中的洗脱情况、缺乏磷酸盐和鲎试剂（LAL）测试结果中看出。纯化后，从枯草芽孢杆菌和化脓性葡萄球菌中提取的LTA能诱导人血液中肿瘤坏死因子α、白细胞介素1 β (il -1 β)、IL-6和IL-10的释放，而从金黄色葡萄球菌中提取的LTA则不能。虽然纯LTA在LAL检测中呈阴性，但在商业制剂中发现超过10 ng LPS/mg LTA的内毒素当量。综上所述，这些内毒素污染相对较高的粗制剂不适合用于表征LTA对免疫细胞的激活作用。
Fractionation of commercial preparations of lipoteichoic acids (LTA) by hydrophobic interaction chromatography (HIC) and nuclear magnetic resonance spectroscopy revealed very inhomogeneous compositions and decomposition of the LTA structure: LTA content of the preparations averaged 61% for Streptococcus pyogenes, 16% for Bacillus subtilis, and 75% for Staphylococcus aureus. The decomposition was characterized by a loss of glycerophosphate units as well as alanine and N-acetylglucosamine substituents. All preparations contained to varying degrees-non-LTA, non-lipopolysaccharide (LPS) immunostimulatory components as indicated by their elution profile in HIC, lack of phosphate, and negative Limulus amoebocyte lysate (LAL) test results. After purification, the commercial LTA from Bacillus subtilis and S. pyogenes but not LTA from S. aureus induced the release of tumor necrosis factor alpha, interleukin I beta (IL-1beta), IL-6, and IL-10 in human blood. While pure LTA are negative in the LAL assay, endotoxin equivalents of more than 10 ng of LPS/mg of LTA were found in the commercial preparations. Taken together, these data indicate that these crude preparations with relatively high endotoxin contamination are not suitable for characterizing the activation of immune cells by LTA.