抗菌肽BNBD5调节肺脏先天免疫抵御胸膜肺炎放线杆菌感染的研究

doi:10.16431/j.cnki.1671-7236.2023.10.040

Abstract

Abstract: 【Objective】 This study was aimed to investigate the role of antimicrobial peptide bovine neutrophil β-defensins 5 (BNBD5) in regulating the innate immune response of lung against Actinobacillus pleuropneumoniae (APP) infection,and provide experimental basis and theoretical support for the prevention and control of porcine pleuropneumonia.【Method】 30 BALB/c mice were randomly divided into three groups,which were normal control group (Con),APP infection group (APP), antibacterial peptide BNBD5 pretreatment and APP infection group (B5),10 mice in each group. Mice in Con group were treated with PBS (20 μL per mouse) two times at two-day intervals. Mice in APP group were treated with PBS (20 μL per mouse) two times at two-day intervals and intranasally challenged with 10⁷ CFU of APP 3 d after treatment. Mice in B5 group were treated with antibacterial peptide BNBD5 (20 μg per mouse) two times at two-day intervals and intranasally challenged with 10⁷ CFU of APP 3 d after treatment.Mice were euthanized 6 h after APP challenge.The general pathological changes of lung and spleen were observed,and the organ coefficient and bacterial load in lung were measured.The pathological changes of lung were observed by HE staining.Levels of cytokines tumor necrosis factor α (TNF-α),interleukin 1β (IL-1β),IL-23,IL-17 and IL-22 in lung were tested using ELISA.The number of neutrophils in lung was measured using flow cytometry.【Result】 Compared with Con group,mice in APP group had rough and disordered hair,decreased mental state,enlarged lungs,and obvious patchy or punctate bleeding.Microscopically,there was significant bleeding and a large amount of inflammatory cell infiltration in lung.The cytokine levels of TNF-α,IL-1β,IL-17,IL-22 and IL-23 in lung were not significantly changed (P＞0.05),but they all decreased to different degrees,and the number of neutrophils decreased.Compared with APP group,the mental state of mice in B5 group was better,the degree of lung swelling was reduced,the lung index was significantly decreased (P＜0.05),no obvious plaque bleeding was observed,the bacterial load in lung was extremely significantly decreased (P＜0.01),and the inflammatory cells in lung were significantly decreased.The levels of TNF-α,IL-1β,IL-17 and IL-22 in lung were significantly increased (P＜0.05),and the number of neutrophils in lung was extremely significantly increased (P＜0.01).【Conclusion】 Antimicrobial peptide BNBD5 could ameliorate APP-caused immune suppression of lung and enhance the innate immunity of lung against APP in the early stages of infection.

Key words: antimicrobial peptide BNBD5; Actinobacillus pleuropneumoniae; immunity of lung; cytokines; neutrophils

CLC Number:

S859

HUANG Jingsheng, ZHU Shuxin, KANG Weichao, DENG Zhijie, YANG Yunmei, LIU Chengzhi, YI Dandan, HE Jiakang, LIANG Zhengmin. Study on Antibacterial Peptide BNBD5 Regulating the Innate Immune Response of Lung Against Actinobacillus pleuropneumoniae[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4261-4269.

References

[1] SASSU E L,BOSSE J T,TOBIAS T J,et al.Update on Actinobacillus pleuropneumoniae-knowledge,gaps and challenges[J].Transboundary and Emerging Diseases,2018,65(S1):72-90.
[2] NAHAR N,TURNI C,TRAM G,et al.Actinobacillus pleuropneumoniae:The molecular determinants of virulence and pathogenesis[J].Advances in Microbial Physiology,2021,78:179-216.
[3] 徐民生,柯海意,施科达,等.广东省猪传染性胸膜肺炎放线杆菌分离鉴定及耐药表型和耐药基因检测与分析[J].中国畜牧兽医,2022,49(8):3212-3225. XU M S,KE H Y,SHI K D,et al.Isolation and identification of Actinobacillus pleuropneumoniae from Guangdong province and detection and analysis of drug resistance phenotype and gene[J]. China Animal Husbandry & Veterinary Medicine,2022,49(8):3212-3225.(in Chinese)
[4] MA X,ZHENG B,WANG J,et al.Quinolone resistance of Actinobacillus pleuropneumoniae revealed through genome and transcriptome analyses[J]. International Journal of Molecular Sciences,2021,22(18):10036.
[5] BAO C,JIANG H,ZHU R,et al.Differences in pig respiratory tract and peripheral blood immune responses to Actinobacillus pleuropneumoniae[J].Veterinary Microbiology,2020,247:108755.
[6] ZHU R,BAO C,LIU B,et al.ITRAQ-based quantitative proteomic analysis of peripheral blood serum in piglets infected with Actinobacillus pleuropneumoniae[J].AMB Express,2020,10(1):121.
[7] BAO C,LIU B,ZHU R,et al.IFN-γ^-/- mice resist Actinobacillus pleuropneumoniae infection by promoting early lung IL-18 release and PMN-Ⅰ accumulation[J].Infection and Immunity,2021,89(6):e00069-21.
[8] MOOKHERJEE N,ANDERSON M A,HAAGSMAN H P,et al.Antimicrobial host defence peptides:Functions and clinical potential[J]. Nature Reviews Drug Discovery,2020,19(5):311-332.
[9] LAZZARO B P,ZASLOFF M,ROLFF J.Antimicrobial peptides:Application informed by evolution[J].Science,2020,368(6490):eaau5480.
[10] MCHUGH B J,WANG R,LI H N,et al.Cathelicidin is a "fire alarm",generating protective NLRP3-dependent airway epithelial cell inflammatory responses during infection with Pseudomonas aeruginosa[J].PLoS Pathogens,2019,15(4):e1007694.
[11] COORENS M,SCHNEIDER V A F,DE GROOT A M,et al.Cathelicidins inhibit Escherichia coli-induced TLR2 and TLR4 activation in a viability-dependent manner[J].Journal of Immunology,2017,199(4):1418-1428.
[12] WANG L,ZHAO X,ZHU C,et al.The antimicrobial peptide MPX kills Actinobacillus pleuropneumoniae and reduces its pathogenicity in mice[J]. Veterinary Microbiology,2020,243:108634.
[13] COORENS M,BANASCHEWSKI B J H,BAER B J,et al.Killing of Pseudomonas aeruginosa by chicken cathelicidin-2 is immunogenically silent,preventing lung inflammation in vivo[J]. Infection and Immunity,2017,85(12):e00546-17.
[14] LIANG Z,LIU Y,SUN X,et al.Immunoregulatory and antimicrobial activity of bovine neutrophil β-defensin-5-loaded PLGA nanoparticles against Mycobacterium bovis[J].Pharmaceutics,2020,12(12):1172.
[15] LIANG Z,LI H,QU M,et al.Intranasal bovine β-defensin-5 enhances antituberculosis immunity in a mouse model by a novel protein-based respiratory mucosal vaccine[J]. Virulence,2022,13(1):949-962.
[16] BAO C T,XIAO J M,LIU B J,et al.Establishment and comparison of Actinobacillus pleuropneumoniae experimental infection model in mice and piglets[J].Microbial Pathogenesis,2019,128:381-389.
[17] WANG L,ZHAO X,ZHU C,et al.Thymol kills bacteria,reduces biofilm formation,and protects mice against a fatal infection of Actinobacillus pleuropneumoniae strain L20[J].Veterinary Microbiology,2017,203:202-210.
[18] CHEN P,BAO C,ZHU R,et al.IL-5 enhances the resistance of Actinobacillus pleuropneumoniae infection in mice through maintaining appropriate levels of lung M2,PMN-Ⅱ and highly effective neutrophil extracellular traps[J].Veterinary Microbiology,2022,269:109438.
[19] YANG X,CHENG Y T,TAN M F,et al.Overexpression of porcine beta-defensin 2 enhances resistance to Actinobacillus pleuropneumoniae infection in pigs[J].Infection and Immunity,2015,83(7):2836-2843.
[20] ZHU R,JIANG H,WANG J,et al.Dynamic immune response characteristics of piglets infected with Actinobacillus pleuropneumoniae through omic[J].AMB Express,2021,11(1):175.
[21] 鲍春彤.胸膜肺炎放线杆菌诱导的肺脏细胞因子应答特征和IL-21作用机制研究[D].长春:吉林大学,2022. BAO C T.Characteristics of pulmonary cytokine response and immunologic mechanism of IL-21 as Actinobacterium pleuropneumoniae infection[D].Changchun:Jilin University,2022.(in Chinese)
[22] CHUEKWON K,CHU C Y,CHENG L T.N-terminus of flagellin enhances vaccine efficacy against Actinobacillus pleuropneumoniae[J]. BMC Veterinary Research,2022,18(1):279.
[23] QI W,ZHU R,BAO C,et al.Porcine circovirus type 2 promotes Actinobacillus pleuropneumoniae survival during coinfection of porcine alveolar macrophages by inhibiting ros production[J].Veterinary Microbiology,2019,233:93-101.
[24] STEHLE C,HERNANDEZ D C,ROMAGNANI C.Innate lymphoid cells in lung infection and immunity[J]. Immunological Reviews,2018,286(1):102-119.
[25] XIONG H,KEITH J W,SAMILO D W,et al.Innate lymphocyte/Ly6C(hi) monocyte crosstalk promotes Klebsiella pneumoniae clearance[J].Cell,2016,165(3):679-689.
[26] MCALEER J P,KOLLS J K.Directing traffic:IL-17 and IL-22 coordinate pulmonary immune defense[J].Immunological Reviews,2014,260(1):129-144.
[27] LIU Y,MEI J,GONZALES L,et al.IL-17A and TNF-α exert synergistic effects on expression of CXCL5 by alveolar type Ⅱ cells in vivo and in vitro[J].Journal of Immunology,2011,186(5):3197-3205.
[28] YANG L,LIU Y,WANG N,et al.Albumin-based LL37 peptide nanoparticles as a sustained release system against Pseudomonas aeruginosa lung infection[J].ACS Biomaterials Science Engineering,2021,7(5):1817-1826.
[29] LIANG W,DIANA J.The dual role of antimicrobial peptides in autoimmunity[J].Frontiers in Immunology,2020,11:2077.
[30] BURN G L,FOTI A,MARSMAN G,et al.The neutrophil[J]. Immunity,2021,54(7):1377-1391.

Study on Antibacterial Peptide BNBD5 Regulating the Innate Immune Response of Lung Against Actinobacillus pleuropneumoniae

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