铜绿假单胞菌重组PcrV-OprI疫苗保护小鼠骨折固定术后感染的作用

张译文; 程新; 张怡; 张月月; 邹全明; 顾江; 肖洪; 胡仁建

doi:10.16016/j.2097-0927.202207117

您当前的位置：

首页 >

文章列表页 >

铜绿假单胞菌重组PcrV-OprI疫苗保护小鼠骨折固定术后感染的作用

基础医学 | 更新时间：2026-01-09

- 铜绿假单胞菌重组PcrV-OprI疫苗保护小鼠骨折固定术后感染的作用
- Recombinant protein vaccine PcrV-OprI prevents mice against Pseudomonas aeruginosa infection after intramedullary nailing
- 陆军军医大学学报 2023年45卷第3期页码：185-191
- 作者机构：
  
  1.400054 重庆, 重庆理工大学药学与生物工程学院
  2.400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系
  3.400020 重庆，陆军军医大学(第三军医大学)958医院骨科
- 作者简介：
  
  [顾江，E-mail：jianggu2012@163.com HU Renjian, E-mail: hrj@cqut.edu.cn
  [胡仁建，E-mail：hrj@cqut.edu.cn GU Jiang, E-mail: jianggu2012@163.com
- 基金信息：
  
  重庆市自然科学基金(cstc2021jcyj-msxmX0799);陆军军医大学临床重大领域技术创新项目(CX2019LC121)
- DOI：10.16016/j.2097-0927.202207117
  中图分类号： R392.7;R619.3;R683.05
- 收稿：2022-07-18，
  
  修回：2022-08-23，
  
  纸质出版：2023-02-15
- 稿件说明：
移动端阅览
张译文, 程新, 张怡, 等. 铜绿假单胞菌重组PcrV-OprI疫苗保护小鼠骨折固定术后感染的作用[J]. 陆军军医大学学报, 2023,45(3):185-191.

Yiwen ZHANG, Xin CHENG, Yi ZHANG, et al. Recombinant protein vaccine PcrV-OprI prevents mice against Pseudomonas aeruginosa infection after intramedullary nailing[J]. Journal of Army Medical University, 2023, 45(3): 185-191.
张译文, 程新, 张怡, 等. 铜绿假单胞菌重组PcrV-OprI疫苗保护小鼠骨折固定术后感染的作用[J]. 陆军军医大学学报, 2023,45(3):185-191. DOI： 10.16016/j.2097-0927.202207117.

Yiwen ZHANG, Xin CHENG, Yi ZHANG, et al. Recombinant protein vaccine PcrV-OprI prevents mice against Pseudomonas aeruginosa infection after intramedullary nailing[J]. Journal of Army Medical University, 2023, 45(3): 185-191. DOI： 10.16016/j.2097-0927.202207117.

摘要

目的

构建髓内针固定小鼠开放性股骨骨折后铜绿假单胞菌(

Pseudomonas aeruginosa

PA)感染模型，评价候选PA疫苗重组PcrV-OprI的免疫保护效果。

方法

采用大肠杆菌表达PcrV-OprI蛋白，依次经亲和、疏水层析纯化得到目的蛋白。C57BL/6J小鼠在第0、14、21天免疫PcrV-OprI蛋白后，使用酶联免疫吸附法检测血清中特异性IgG抗体及亚型效价。末次免疫后14 d，进行髓内针固定小鼠开放性股骨骨折术。术后PAO1菌株感染，通过比较免疫组与对照组细菌定植、体质量、组织病理学和X线影像表现等差异，评价PcrV-OprI抗原保护效果。

结果

PcrV-OprI免疫组小鼠血清中特异性抗体水平显著升高(

0.05)，且以IgG1亚型为主；PcrV-OprI免疫组体质量减轻幅度显著低于对照组(

0.05)，PcrV-OprI免疫组软组织、骨内、植入物细菌数量显著低于对照组(

0.05)；PcrV-OprI免疫组炎症病理损伤小于对照组；在观察期结束时，PcrV-OprI免疫组骨折线清晰且逐渐闭合，呈愈合趋势而对照组骨折处未见明显愈合。

结论

PcrV-OprI免疫能有效引起小鼠体液免疫应答，在开放性骨折髓内针固定术后PA感染模型中具有显著保护效果。

Abstract

Objective

To establish a model of

Pseudomonas aeruginosa

(PA) infection in mice with open fractures after the femur with intramedullary nailing and to explore the immunoprotective effects of recombinant PcrV-OprI.

Methods

PcrV-OprI protein was expressed in

E. coli

and then the target protein was purified by affinity chromatography and hydrophobic interaction chromatography. The C57BL/6J mice were immunized with PcrV-OprI protein on Day 0

14 and 21. After that

the titers of specific IgGs and their subtypes were determined by ELISA. In 14 d after the final vaccination

open femur fractures and intramedullary nailing were performed in mice

which was followed by a challenge with PAO1 strain. The protection of PcrV-OprI was assessed with bacterial colonization

body weight

histology

and X-ray imaging.

Results

The levels of specific antibodies were significantly higher in PcrV-OprI-immunized sera than that of the control group (

0.05)

and the obtained IgG1 was the predominating subtype. The loss of body weight of the immunized mice was significantly lower than that of the control group (

0.05). In addition

a similiar trend was observed in the amount of colonized bacteria in the tissues

bones

and implants (

0.05). Moreover

the histological observation suggested that bacteria caused less damage in the PcrV-OprI-immunized mice. At the end of the observation period

the fracture line in immunized mice was clear

indicating gradual healing.

Conclusion

Vaccination of PcrV-OprI is efficient in causing a humoral immune response and shows significant protective effect on PA infection model in mice after open fractures of the femur with intramedullary nailing.

关键词

Keywords

references

BASSETTI M, VENA A, CROXATTO A, et al . How to manage Pseudomonas aeruginosa infections[J ] . Drugs Context, 2018, 7: 212527. DOI:10.7573/dic.212527..

TRIFFAULT-FILLIT C, FERRY T, LAURENT F, et al . Microbiologic epidemiology depending on time to occurrence of prosthetic joint infection: a prospective cohort study[J ] . Clin Microbiol Infect, 2019, 25(3): 353-358. DOI:10.1016/j.cmi.2018.04.035.

CHEVALIER S, BOUFFARTIGUES E, BODILIS J, et al . Structure, function and regulation of Pseudomonas aeruginosa porins[J ] . FEMS Microbiol Rev, 2017, 41(5): 698-722. DOI:10.1093/femsre/fux020..

PELEGRIN A C, PALMIERI M, MIRANDE C, et al . Pseudomonas aeruginosa : a clinical and genomics update[J ] . FEMS Microbiol Rev, 2021, 45(6): fuab026. DOI:10.1093/femsre/fuab026..

HOGGARTH A, WEAVER A, PU Q Q, et al . Mechanistic research holds promise for bacterial vaccines and phage therapies for Pseudomonas aeruginosa [J ] . Drug Des Devel Ther, 2019, 13: 909-924. DOI:10.2147/DDDT.S189847..

THANABALASURIAR A, SUREWAARD B G, WILLSON M E, et al . Bispecific antibody targets multiple Pseudomonas aeruginosa evasion mechanisms in the lung vasculature[J ] . J Clin Invest, 2017, 127(6): 2249-2261. DOI:10.1172/JCI89652..

BORKAR D S, FLEISZIG S M, LEONG C, et al . Association between cytotoxic and invasive Pseudomonas aeruginosa and clinical outcomes in bacterial keratitis[J ] . JAMA Ophthalmol, 2013, 131(2): 147-153. DOI:10.1001/jamaophthalmol.2013.778..

FELTMAN H, SCHULERT G, KHAN S, et al . Prevalence of type Ⅲ secretion genes in clinical and environmental isolates of Pseudomonas aeruginosa [J ] . Microbiology (Reading), 2001, 147(Pt 10): 2659-2669. DOI:10.1099/00221287-147-10-2659..

GAREY K W, VO Q P, LAROCCO M T, et al . Prevalence of type Ⅲ secretion protein exoenzymes and antimicrobial susceptibility patterns from bloodstream isolates of patients with Pseudomonas aeruginosa bacteremia[J ] . J Chemother, 2008, 20(6): 714-720. DOI:10.1179/joc.2008.20.6.714..

WAN C, GAO C, XIE Q, et al . Flagella hook protein FlgE is a novel vaccine candidate of Pseudomonas aeruginosa identified by a genomic approach[J ] . Vaccine, 2021, 39(17): 2386-2395. DOI:10.1016/j.vaccine.2021.03.051..

刘潇, 李文桂. 铜绿假单胞菌外膜蛋白I(OprI)疫苗的研制现状[J]. 中国病原生物学杂志, 2017, 12(6): 590-595. DOI:10.13350/j.cjpb.170625.

LIU X, LI W G. Status of research on the outer membrane protein I of Pseudomonas aeruginosa [J ] . J Pathog Biol, 2017, 12(6): 590-595. DOI:10.13350/j.cjpb.170625.

OEZEL L, BVREN C, SCHOLZ A O, et al . Effect of antibiotic infused calcium sulfate/hydroxyapatite (CAS/HA) insets on implant-associated osteitis in a femur fracture model in mice[J ] . PLoS One, 2019, 14(3): e0213590. DOI:10.1371/journal.pone.0213590.

GU J, LIU Y Q, YU S, et al . Enterohemorrhagic Escherichia coli trivalent recombinant vaccine containing EspA, intimin and Stx2 induces strong humoral immune response and confers protection in mice[J ] . Microbes Infect, 2009, 11(10/11): 835-841. DOI:10.1016/j.micinf.2009.04.024..

GU H, LIAO Y L, ZHANG J, et al . Rational design and evaluation of an artificial Escherichia coli K1 protein vaccine candidate based on the structure of OmpA[J ] . Front Cell Infect Microbiol, 2018, 8: 172. DOI:10.3389/fcimb.2018.00172..

LUO M, SHAO B, YU J Y, et al . Simultaneous enhancement of cellular and humoral immunity by the high salt formulation of Al(OH)3 adjuvant[J ] . Cell Res, 2017, 27(4): 586-589. DOI:10.1038/cr.2017.14.

WANG Y, CHENG X, WAN C, et al . Development of a chimeric vaccine against Pseudomonas aeruginosa based on the Th17-stimulating epitopes of PcrV and AmpC[J ] . Front Immunol, 2020, 11: 601601. DOI:10.3389/fimmu.2020.601601..

SAWA T, ITO E, NGUYEN V H, et al . Anti-PcrV antibody strategies against virulent Pseudomonas aeruginosa [J ] . Hum Vaccin Immunother, 2014, 10(10): 2843-2852. DOI:10.4161/21645515.2014.971641..

WARRENER P, VARKEY R, BONNELL J C, et al . A novel anti-PcrV antibody providing enhanced protection against Pseudomonas aeruginosa in multiple animal infection models[J ] . Antimicrob Agents Chemother, 2014, 58(8): 4384-4391. DOI:10.1128/AAC.02643-14..

MEYER S, LEUSEN J H W, BOROSS P. Regulation of complement and modulation of its activity in monoclonal antibody therapy of cancer[J]. mAbs, 2014, 6(5): 1133-1144. DOI:10.4161/mabs.29670.

WONG R M Y, LI T K, LI J, et al . A systematic review on current osteosynthesis-associated infection animal fracture models[J ] . J Orthop Translat, 2020, 23: 8-20. DOI:10.1016/j.jot.2020.03.002.

THOMPSON J M, MILLER R J, ASHBAUGH A G, et al . Mouse model of Gram-negative prosthetic joint infection reveals therapeutic targets[J ] . JCI Insight, 2018, 3(17): 121737. DOI:10.1172/jci.insight.121737.

SHIELS S M, BEDIGREW K M, WENKE J C. Development of a hematogenous implant-related infection in a rat model[J]. BMC Musculoskelet Disord, 2015, 16: 255. DOI:10.1186/s12891-015-0699-7.

HALL-STOODLEY L, STOODLEY P. Evolving concepts in biofilm infections[J]. Cell Microbiol, 2009, 11(7): 1034-1043. DOI:10.1111/j.1462-5822.2009.01323.x.

SHORTRIDGE D, PFALLER M A, STREIT J M, et al . Antimicrobial activity of ceftolozane/tazobactam tested against contemporary (2015-2017) Pseudomonas aeruginosa isolates from a global surveillance programme[J ] . J Glob Antimicrob Resist, 2020, 21: 60-64. DOI:10.1016/j.jgar.2019.10.009..

BASSETTI M, VENA A, CROXATTO A, et al . How to manage Pseudomonas aeruginosa infections[J ] . Drugs Context, 2018, 7: 212527. DOI:10.7573/dic.212527..

BURROWS L L. The therapeutic pipeline for Pseudomonas aeruginosa infections[J ] . ACS Infect Dis, 2018, 4(7): 1041-1047. DOI:10.1021/acsinfecdis.8b00112.

NGUYEN L, GARCIA J, GRUENBERG K, et al . Multidrug-resistant Pseudomonas infections: hard to treat, but hope on the horizon? [J ] . Curr Infect Dis Rep, 2018, 20(8): 23. DOI:10.1007/s11908-018-0629-6.

PARKINS M D, SOMAYAJI R, WATERS V J. Epidemiology, biology, and impact of clonal Pseudomonas aeruginosa infections in cystic fibrosis[J ] . Clin Microbiol Rev, 2018, 31(4): e00019-e00018. DOI:10.1128/CMR.00019-18.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

铜绿假单胞菌噬菌体vB_PaS_IME307的生物学特性研究及全基因组分析

信号分子AI-2通过密度感应系统促进铜绿假单胞菌生物被膜形成和毒力因子产生

光动力抗菌疗法对多重耐药铜绿假单胞菌体外杀伤作用的研究

小鼠铜绿假单胞菌气管插管滴注法肺部感染模型的建立及评价

铜绿假单胞菌噬菌体PA-27-1的分离及其生物学特性