毒力相关染色体位点J (VacJ)是一种外膜脂蛋白,可引起小鼠对鲍曼不动杆菌感染的保护性免疫

Saeideh Masoumkhani, S. D. A. Astaneh, Abolfazl Jahangiri, I. Rasooli
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引用次数: 2

摘要

鲍曼不动杆菌是革兰氏阴性、严格需氧非运动细菌,其DNA G-C含量为39%至47%。鲍曼不动杆菌是一种重要的机会性医院病原体,可引起肺炎、菌血症、尿路感染、脑膜炎、皮肤和软组织感染。鲍曼不动杆菌的主要问题是耐药菌株的出现,这需要开发新的预防、控制和治疗方法。近年来,细菌脂蛋白在诱导机体对传染病产生保护性免疫方面受到了研究人员的关注。VacJ是一种高度保守的外膜脂蛋白,存在于许多鲍曼不动杆菌菌株中,值得对其免疫原性进行研究。克隆了鲍曼不动杆菌成熟vacJ编码基因ATCC19606,并以融合蛋白的形式在大肠杆菌中过表达。重组VacJ (rVacJ)得到纯化。20µg和40µg纯化的~ 31 kDa rVacJ与Freund或Alum佐剂一起用于小鼠免疫。间接ELISA法测定重组蛋白的抗体效价。以1:200血清稀释法检测鲍曼不动杆菌全细胞。用不同剂量的鲍曼不动杆菌对小鼠进行主动、被动和鼻内感染。测定对照组和免疫组小鼠肺部的细菌负荷。高抗体滴度被注意到是免疫rVacJ的结果。与rVacJ-Alum佐剂相比,rVacJ-Freund佐剂激发了更高的抗体水平。腹腔内注射活鲍曼不动杆菌的小鼠组没有存活,而鼻内注射组肺部细菌负荷显著减少了600倍。这些发现对于开发新型佐剂疫苗和精确的给药路线以对抗臭名昭著的多药耐药鲍曼杆菌具有重要价值。
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Virulence-associated chromosome locus J, VacJ, an outer membrane lipoprotein elicits protective immunity against Acinetobacter baumannii infection in mice
Acinetobacter baumannii is a Gram-negative, strictly aerobic nonmotile bacterium with a DNA G-C content of 39% to 47%. A. baumannii is an important opportunistic nosocomial pathogen that causes pneumonia, bacteremia, urinary tract infections, meningitis, skin, and soft tissue infections. The major concern about A.baumannii is the emergence of resistant strains which necessitates the development of new prevention, control, and treatment methods. Recently the bacterial lipoproteins have attracted the attention of the researchers for the induction of protective immunity against infectious diseases. VacJ is a highly conserved outer membrane lipoprotein that exists in many A. baumannii strains that deserves research on its immunogenicity. The gene encoding mature vacJ of A. baumannii ATCC19606 was cloned and over-expressed in Escherichia coli as a fusion protein. The recombinant VacJ (rVacJ) was purified. 20 and 40 µg of the purified ∼ 31 kDa rVacJ were used for immunization of mice along with Freund’s or Alum adjuvants. Antibody titres raised against the recombinant protein were determined by indirect ELISA. Whole A.baumannii cell was detected at 1:200 serum dilution. Bacterial challenges of mice groups with varying doses of A. baumannii were performed in the active, passive, and intranasal forms. The bacterial load in the mice lungs was determined in both control and immunized groups. A high antibody titre was noted as a result of immunization with rVacJ. The rVacJ-Freund’s adjuvant elicited a higher antibody level compared to the rVacJ-Alum adjuvant. The mice groups challenged intraperitoneally with live A. baumannii did not survive while the intranasally challenged group exhibited a significant reduction of 600 fold of the bacterial load in the lungs. The findings are of significant value in the development of novel adjuvanted vaccines and precise routes of administrations in combat against the notorious multidrug-resistant A.bauamnnii .
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