Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India.
Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India.
Microb Pathog. 2021 Sep;158:104974. doi: 10.1016/j.micpath.2021.104974. Epub 2021 May 18.
Probiotic intervention has been long believed to have beneficial effects on human health by curbing the intestinal colonization of pathogens. However, the application of live probiotics therapy may not be an ideal approach to circumvent the infections of superbug origin due to the risk of horizontal antibiotic resistance genes transfer. In this study, the anti-adhesion ability of extractable cell surface proteins from two indigenous potential probiotic strains (Lactiplantibacillus plantarum A5 and Limosilactobacillus fermentum Lf1) and two standard reference strains (Lactobacillus acidophilus NCFM and Lacticaseibacillus rhamnosus LGG) was evaluated against clinical isolates of Methicillin-Resistant Staphylococcus aureus (MRSA) on porcine gastric mucin and HT-29 cells. The surface proteins from the probiotic strains were extracted by treatment with 5 M lithium chloride. The surface protein quantification and SDS-PAGE profiling indicated that the yield and protein patterns were strain-specific. Surface proteins significantly hampered the mucoadhesion of MRSA isolates via protective, competitive, and displacement. Similarly, the treatment with surface proteins probiotic strains displayed anti-adhesion against MRSA isolates on HT-29 cells without affecting the viability of the cell line. Surface proteins treatment to the confluent monolayer of HT-29 cells maintained the epithelial integrity; however, MRSA isolates (10 cells/mL) showed considerable alteration in the epithelial integrity by exacerbating the FITC-dextran transflux. Contrarily, the co-treatment with surface proteins with MRSA isolates significantly lowered the FITC-dextran transflux across the differentiated HT-29 monolayer. Overall, the findings of this study suggest that probiotic-derived surface proteins could be the novel biotherapeutics to combat the MRSA colonization and their concomitant intestinal infections.
益生菌干预被长期认为通过抑制病原体在肠道的定植对人类健康有益。然而,由于水平抗生素耐药基因转移的风险,活益生菌治疗的应用可能不是规避超级细菌来源感染的理想方法。在这项研究中,评估了从两个本土潜在益生菌株(植物乳杆菌 A5 和发酵乳杆菌 Lf1)和两个标准参考株(嗜酸乳杆菌 NCFM 和鼠李糖乳杆菌 LGG)中提取的可提取细胞表面蛋白对猪胃粘蛋白和 HT-29 细胞上耐甲氧西林金黄色葡萄球菌(MRSA)临床分离株的抗粘附能力。通过用 5M 氯化锂处理提取益生菌株的表面蛋白。表面蛋白定量和 SDS-PAGE 分析表明,产率和蛋白模式具有菌株特异性。表面蛋白通过保护、竞争和置换显着阻碍了 MRSA 分离株的粘膜粘附。同样,表面蛋白益生菌株处理对 HT-29 细胞上的 MRSA 分离株显示出抗粘附作用,而不影响细胞系的活力。表面蛋白处理到 HT-29 细胞的汇合单层维持上皮完整性;然而,MRSA 分离株(10 个细胞/毫升)通过加剧 FITC-葡聚糖通量来显著改变上皮完整性。相反,用表面蛋白与 MRSA 分离株共同处理可显著降低穿过分化的 HT-29 单层的 FITC-葡聚糖通量。总的来说,这项研究的结果表明,益生菌衍生的表面蛋白可能是对抗 MRSA 定植及其伴随的肠道感染的新型生物治疗剂。