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Monoclonal antibodies specific to streptococcus mutans GS-5 glucosyltransferase-C inhibit bacterial glucosyltransferase

机译:变形链球菌GS-5葡萄糖基转移酶C特异的单克隆抗体抑制细菌葡萄糖基转移酶

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摘要

Glucosyltransferase-C (GTFC) is a virulence factor of Streptococcus mutans. Additionally, GTFC represents an essential element required for bacterial cell coherence, allowing for the formation of dental plaque, which leads to dental caries. As such, monoclonal antibodies (MAbs) against S. mutans are believed to offer some protection against dental caries. In the current study, we amplified an approximately 1.5 kb fragment of the N-terminal half of the S. mutans gtfC gene by PCR, then induced expression of this gene. This protein was designated GTFCN. After the expressed protein was purified, it was used as an immunogen and injected into BALB/c mice. We selected and established two MAbs by producing hybridomas (HCN17 and HCN37). The anti-GTFCN antibody isotype was confirmed as IgG2a for HCN17 and IgG2b for HCN37. The anti-GTFCN antibody was found to specifically react with the GTFCN protein. The enzymatic activity of the crude glucosyltransferase of S. mutans GS-5 was significantly inhibited at a concentration of 350 ng of MAb/mL. These results suggest that the monoclonal anti-GTFCN antibodies could represent an alternative modality for passive immunization to prevent S. mutans aggregation and dental plaque.
机译:葡萄糖基转移酶-C(GTFC)是变形链球菌的致病因子。此外,GTFC代表细菌细胞凝聚所需的必要元素,可形成牙菌斑,从而导致龋齿。这样,据信抗变形链球菌的单克隆抗体(MAbs)提供了一些抗龋齿的保护。在当前的研究中,我们通过PCR扩增了变形链球菌gtfC基因N端大约1.5 kb的片段,然后诱导该基因的表达。该蛋白称为GTFCN。纯化表达的蛋白质后,将其用作免疫原并注射到BALB / c小鼠中。我们通过产生杂交瘤(HCN17和HCN37)选择并建立了两个单克隆抗体。确认抗GTFCN抗体同种型为HCN17的IgG2a和HCN37的IgG2b。发现抗GTFCN抗体与GTFCN蛋白特异性反应。在浓度为350 ng MAb / mL的情况下,变形链球菌GS-5的粗糖基转移酶的酶活性受到明显抑制。这些结果表明,单克隆抗GTFCN抗体可以代表被动免疫的另一种形式,以防止变形链球菌聚集和牙菌斑。

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