Department of Biology, Shahed University, Tehran-Qom Express Way, Iran.
Department of Biology, Shahed University, Tehran-Qom Express Way, Iran; Molecular Microbiology Research Center, Shahed University, Tehran-Qom Express Way, Iran.
Microb Pathog. 2021 Dec;161(Pt B):105291. doi: 10.1016/j.micpath.2021.105291. Epub 2021 Nov 17.
Acinetobacter baumannii, an opportunistic extracellular pathogen is one of the major causes of nosocomial infections. Omp34, also known as Omp33-36, is a bacterial porin protein involved in the virulence and fitness of this pathogen by adhesion to the host cell. This antigen nominated as an appropriate candidate for immunization against A. baumannii. In this study, the expression of the recombinant Omp34 (rOmp34) was carried out in E. coli BL21 (DE3). The immunogenicity of the rOmp34 in A. baumannii was studied in a murine sepsis model. Antibody response in mice injected with the recombinant protein was assessed using indirect ELISA. Bactericidal activity of rOmp34-immunized mice sera (1:10 dilution) against A. baumannii ATCC 19606 after 0, 1, 2, 4, and 8 h of incubation at 37 °C was assessed. In addition to survival rate, load of bacteria in liver and spleen of the infected mice were evaluated. A high titer of specific antibody equivalent to optical density of 1.54 ± 0.06 against rOmp34 was elicited in the immunized mice sera. Viability of the A. baumannii incubated 8 h with immunized mice sera was 64%. Homogenized liver and spleen samples of the control mice challenged with A. baumannii were loaded with 8 × 10 and 9 × 10 CFU per gram tissue respectively 48 h post-challenge as against complete clearance of A. baumannii in the immunized group. The protective immunity was achieved by challenging the mice groups with 5 × LD of live A. baumannii. Omp34 can be nominated as an immunogen that can bring about protection against Acinetobacter baumannii.
鲍曼不动杆菌是一种机会性细胞外病原体,是医院感染的主要原因之一。Omp34,也称为 Omp33-36,是一种细菌孔道蛋白,通过与宿主细胞黏附参与该病原体的毒力和适应性。这种抗原被提名作为针对鲍曼不动杆菌免疫接种的合适候选物。在这项研究中,在大肠杆菌 BL21(DE3)中进行了重组 Omp34(rOmp34)的表达。在小鼠败血症模型中研究了 rOmp34 在鲍曼不动杆菌中的免疫原性。通过间接 ELISA 评估注射重组蛋白的小鼠中的抗体反应。在 37°C 孵育 0、1、2、4 和 8 小时后,评估 rOmp34 免疫小鼠血清(1:10 稀释)对 ATCC 19606 鲍曼不动杆菌的杀菌活性。除了存活率外,还评估了感染小鼠肝脏和脾脏中的细菌负荷。在免疫小鼠血清中诱导了针对 rOmp34 的高滴度特异性抗体,相当于 1.54±0.06 的光密度。与对照小鼠相比,用免疫小鼠血清孵育 8 小时的鲍曼不动杆菌的存活率为 64%。在挑战后 48 小时,用鲍曼不动杆菌挑战的对照小鼠的肝和脾匀浆样本分别载有 8×10 和 9×10 CFU/克组织,而在免疫组中完全清除了鲍曼不动杆菌。通过用 5×LD 的活鲍曼不动杆菌挑战小鼠组来实现保护免疫力。Omp34 可以被提名作为一种免疫原,可带来针对鲍曼不动杆菌的保护。