Eghbal M, Khaki P, Ghandehari F, Taghizadeh M, Tebianian M
Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran.
Department of Microbiology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
Arch Razi Inst. 2024 Oct 31;79(5):1099-1108. doi: 10.32592/ARI.2024.79.5.1099. eCollection 2024 Oct.
Leptospirosis represents a significant threat to the health of both humans and animals. It is a disease caused by pathogenic Leptospira. The early detection of pathogenic Leptospira is an effective method for preventing a multitude of potential complications. The protected outer membrane protein (OMP) of pathogenic Leptospira, LipL41-OmpL1, was inserted into E. coli bacteria using different software for the amino acid sequence of OmpL1 and LipL41. This was done to design a recombinant fusion protein, which was then expressed to investigate immunogenicity. The selected genes were propagated and cloned as a fusion in a PET32a+ plasmid vector and expressed by Escherichia coli strain S (DE3) via a heat shock method. The evaluation was conducted using the BALB/c mouse as the laboratory animal model. The recombinant LipL41-OmpL1 protein was confirmed using the urea purification method and western blot, and its immunogenicity was evaluated by measuring the high humoral immune stimulation and antibody secretion in BALB/c mice by the ELISA method. The findings demonstrated that the animals that received both the OmpL1 and LipL41 proteins exhibited 85% immunogenicity, whereas the control group that did not receive the fusion protein demonstrated only 25% immunogenicity (P<0.001). Moreover, no evidence of infection was identified in recipients of the OmpL1-LipL41 fusion protein, indicating that this protein is safe for use. The protective effects of immunization with OmpL1 and LipL41 were synergistic, as no significant levels of protection were observed in animals immunized with OmpL1 or LipL41 alone. In conclusion, this study underscores the potential of a recombinant OmpL1 and LipL41 fusion protein as a promising avenue for research in the development of vaccines and ELISA diagnostic kits for the prevention and rapid diagnosis of leptospirosis.
钩端螺旋体病对人类和动物的健康构成重大威胁。它是由致病性钩端螺旋体引起的疾病。早期检测致病性钩端螺旋体是预防多种潜在并发症的有效方法。利用不同软件根据OmpL1和LipL41的氨基酸序列,将致病性钩端螺旋体的保护性外膜蛋白(OMP)LipL41 - OmpL1插入大肠杆菌中。这样做是为了设计一种重组融合蛋白,然后将其表达以研究免疫原性。所选基因在PET32a +质粒载体中作为融合体进行扩增和克隆,并通过热休克法由大肠杆菌菌株S(DE3)进行表达。以BALB / c小鼠作为实验动物模型进行评估。使用尿素纯化法和蛋白质印迹法对重组LipL41 - OmpL1蛋白进行鉴定,并通过ELISA法测量BALB / c小鼠中的高体液免疫刺激和抗体分泌来评估其免疫原性。研究结果表明,同时接受OmpL1和LipL41蛋白的动物表现出85%的免疫原性,而未接受融合蛋白的对照组仅表现出25%的免疫原性(P<0.001)。此外,在接受OmpL1 - LipL41融合蛋白的动物中未发现感染迹象,表明该蛋白使用安全。单独用OmpL1或LipL41免疫的动物未观察到显著的保护水平,而用OmpL1和LipL41免疫的保护作用具有协同性。总之,本研究强调了重组OmpL1和LipL41融合蛋白在开发用于预防和快速诊断钩端螺旋体病的疫苗和ELISA诊断试剂盒方面作为一种有前景的研究途径的潜力。