Dehnavi Meghdad, Haghighat Setareh, Yazdi Mohammad Hossein, Mahdavi Mehdi
Department of Microbiology, Faculty of Pharmaceutical Sciences, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Department of Microbiology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Microb Pathog. 2023 Mar;176:106007. doi: 10.1016/j.micpath.2023.106007. Epub 2023 Jan 26.
Staphylococcus aureus is a gram-positive bacterium, representing one of the most important nosocomial pathogens. The treatment of infections, caused by S. aureus, has become increasingly intricate due to the emergence of highly resistant strains. Therefore, it is obvious that an effective prevention strategy against this bacterium could significantly decrease such infections. In the present study, the protective efficacy and immunological properties of recombinant autolysin, formulated in Montanide ISA266 and Alum adjuvants with Glucomannan as a polysaccharide, were assessed in the systemic mouse model of infection. Mice were immunized with the purified recombinant protein in various formulations in different groups and, subsequently, mice were challenged with 5 × 10 CFU of bacteria for the evaluation of their survival and bacterial clearances in the internal organs. ELISA was performed to determine the type of induced immunity, cytokine secretion (IFN-γ, IL-4, IL-2, and IL-17), and isotyping (IgG1 and IgG2a). In addition, we measured the opsonophagocytic activities of the antibodies. Results showed that immunization with r-autolysin + Alum + Glucomannan and r-autolysin + MontanideISA266+Glucomannan formulations significantly increased total IgG and isotypes (IgG1 and IgG2a), as compared with other vaccinated and control groups. Furthermore, the formulation of r-autolysin in Alum and MontanideISA266 adjuvants with Glucomannan enhanced IFN-γ, IL-4, and IL-17 cytokine secretion as well as protectivity, following experimental challenge. We concluded that Glucomannan has the potential to induce immune responses and would be used as an adjuvant factor in vaccine formulation.
金黄色葡萄球菌是一种革兰氏阳性细菌,是最重要的医院病原体之一。由于高耐药菌株的出现,由金黄色葡萄球菌引起的感染治疗变得越来越复杂。因此,显然针对这种细菌的有效预防策略可以显著减少此类感染。在本研究中,在小鼠全身感染模型中评估了以Montanide ISA266和明矾为佐剂、以葡甘露聚糖为多糖配制的重组自溶素的保护效力和免疫特性。将不同组的小鼠用各种制剂中的纯化重组蛋白进行免疫,随后用5×10⁶CFU细菌攻击小鼠,以评估其存活率和内脏中的细菌清除情况。进行ELISA以确定诱导免疫的类型、细胞因子分泌(IFN-γ、IL-4、IL-2和IL-17)和亚型(IgG1和IgG2a)。此外,我们测量了抗体的调理吞噬活性。结果表明,与其他接种组和对照组相比,用重组自溶素+明矾+葡甘露聚糖和重组自溶素+Montanide ISA266+葡甘露聚糖制剂免疫显著增加了总IgG和亚型(IgG1和IgG2a)。此外,在明矾和Montanide ISA266佐剂中用葡甘露聚糖配制重组自溶素,在实验攻击后增强了IFN-γ、IL-4和IL-17细胞因子的分泌以及保护作用。我们得出结论,葡甘露聚糖有诱导免疫反应的潜力,可作为疫苗制剂中的佐剂因子。