Granato D, Perotti F, Masserey I, Rouvet M, Golliard M, Servin A, Brassart D
Nestlé Research Center, CH-1000 Lausanne 26, Switzerland.
Appl Environ Microbiol. 1999 Mar;65(3):1071-7. doi: 10.1128/AEM.65.3.1071-1077.1999.
The influence of pH on the adhesion of two Lactobacillus strains to Caco-2 human intestinal cells was investigated. One strain, Lactobacillus johnsonii La1, was adherent at any pH between 4 and 7. The other one, L. acidophilus La10, did not attach to this cell line under the same experimental conditions. On the basis of these results, we used the monoclonal antibody technique as a tool to determine differences on the surface of these bacteria and to identify a factor for adhesion. Mice were immunized with live La1, and the hybridomas produced by fusion of spleen cells with ONS1 cells were screened for the production of antibodies specific for L. johnsonii La1. A set of these monoclonal antibodies was directed against a nonproteinaceous component of the L. johnsonii La1 surface. It was identified as lipoteichoic acid (LTA). This molecule was isolated, chemically characterized, and tested in adhesion experiments in the same system. The adhesion of L. johnsonii La1 to Caco-2 cells was inhibited in a concentration-dependent way by purified LTA as well as by L. johnsonii La1 culture supernatant that contained LTA. These results showed that the mechanism of adhesion of L. johnsonii La1 to human Caco-2 cells involves LTA.
研究了pH对两种乳酸杆菌菌株黏附于Caco-2人肠道细胞的影响。其中一株约氏乳杆菌La1在pH值4至7之间的任何条件下均具有黏附性。另一株嗜酸乳杆菌La10在相同实验条件下不黏附于该细胞系。基于这些结果,我们使用单克隆抗体技术作为工具来确定这些细菌表面的差异,并鉴定黏附因子。用活的La1免疫小鼠,筛选由脾细胞与ONS1细胞融合产生的杂交瘤,以获得对约氏乳杆菌La1具有特异性的抗体。一组这些单克隆抗体针对约氏乳杆菌La1表面的一种非蛋白质成分。它被鉴定为脂磷壁酸(LTA)。分离出该分子,进行化学表征,并在同一系统的黏附实验中进行测试。纯化的LTA以及含有LTA的约氏乳杆菌La1培养上清液以浓度依赖的方式抑制约氏乳杆菌La1对Caco-2细胞的黏附。这些结果表明,约氏乳杆菌La1黏附于人Caco-2细胞的机制涉及LTA。