Top

Published in:

Open Access 01-12-2017 | Research article

Recombinant Salmonella expressing SspH2-EscI fusion protein limits its colonization in mice

Authors: Maozhi Hu, Weixin Zhao, Wei Gao, Wenhua Li, Chuang Meng, Qiuxiang Yan, Yuyang Wang, Xiaohui Zhou, Shizhong Geng, Zhiming Pan, Guiyou Cui, Xinan Jiao

Published in: BMC Immunology | Issue 1/2017

Abstract

Background

Activation of inflammasome contributes to the clearance of intracellular bacteria. C-terminus of E. coli EscI protein can activate NLRC4 (NLR family, CARD domain containing-4) inflammasome in macrophages. The purpose of this study was to determine if activation of NLRC4 inflammasome by EscI can reduce the colonization of Salmonella in mice.

Results

A recombinant S. typhimurium strain expressing fusion protein of the N-terminal SspH2 (a Salmonella type III secretion system 2 effector) and C-terminal EscI was constructed and designated as X4550(pYA3334-SspH2-EscI). In vitro assay showed that X4550(pYA3334-SspH2-EscI) significantly enhanced IL-1β and IL-18 secretion (P < 0.05) and pyroptotic cell death of mouse peritoneal macrophages, compared with those infected with control strain, X4550(pYA3334-SspH2). In vivo studies showed that colonization of X4550(pYA3334-SspH2-EscI) in both spleen and liver were significantly lower than that of X4550(pYA3334-SspH2) (P < 0.05). The bacterial counts of X4550(pYA3334-SspH2-EscI) in mice decreased, while those of X4550(pYA3334-SspH2) increased over the time after infection. Additionally, X4550(pYA3334-SspH2-EscI) induced a less pathological alteration in spleen and liver than X4550(pYA3334-SspH2).

Conclusion

Fusion protein SspH2-EscI may be translocated into macrophages and activate NLRC4 inflammasome, which limits Salmonella colonization in spleen and liver of mice.

Skeldon A, Saleh M. The inflammasomes: molecular effectors of host resistance against bacterial, viral, parasitic, and fungal infections. Front Microbiol. 2011;2:15.CrossRefPubMedPubMedCentral

Kofoed EM, Vance RE. Innate immune recognition of bacterial ligands by NAIPs dictates inflammasome specificity. Nature. 2012;477(7366):592–5.CrossRef

Monie TP, Bryant CE, Gay NJ. Activating immunity: lessons from the TLRs and NLRs. Trends Biochem Sci. 2009;34(11):553–61.CrossRefPubMed

Khameneh HJ, Mortellaro A. NLRC4 gets out of control. Nat Genet. 2014;46(10):1048–9.CrossRefPubMed

Broz P, Newton K, Lamkanfi M, Mariathasan S, Dixit VM, Monack DM. Redundant roles for inflammasome receptors NLRP3 and NLRC4 in host defense against Salmonella. J Exp Med. 2010;207(8):1745–55.CrossRefPubMedPubMedCentral

Carvalho FA, Nalbantoglu I, Aitken JD, Uchiyama R, Su Y, Doho GH, et al. Cytosolic flagellin receptor NLRC4 protects mice against mucosal and systemic challenges. Mucosal Immunol. 2012;5(3):288–98.CrossRefPubMedPubMedCentral

Zhao Y, Yang J, Shi J, Gong YN, Lu Q, Xu H, et al. The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature. 2011;477(7366):596–600.CrossRefPubMed

Miao EA, Leaf IA, Treuting PM, Mao DP, Dors M, Sarkar A, et al. Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria. Nat Immunol. 2010;11(12):1136–42.CrossRefPubMedPubMedCentral

Stevens MP, Humphrey TJ, Maskell DJ. Molecular insights into farm animal and zoonotic Salmonella infections. Phil Trans R Soc B. 2009;364:2709–23.CrossRefPubMedPubMedCentral

10.

Lara-Tejero M, Kato J, Wagner S, Liu X, Galán JE. A Sorting platform determines the order of protein secretion in bacterial type III systems. Science. 2011;331:1188–91.CrossRefPubMed

11.

Yu XJ, McGourty K, Liu M, Unsworth KE, Holden DW. pH sensing by intracellular Salmonella induces effector translocation. Science. 2010;328(5981):1040–43.CrossRefPubMed

12.

McGhie EJ, Brawn LC, Hume PJ, Humphreys D, Koronakis V. Salmonella takes control: effector-driven manipulation of the host. Curr Opin Microbiol. 2009;12(1):117–24.CrossRefPubMedPubMedCentral

13.

Lahiri A, Lahiri A, Iyer N, Das P, Chakravortty D. Visiting the cell biology of Salmonella infection. Microbes Infect. 2010;12:809–18.CrossRefPubMed

14.

Miao EA, Warren SE. Innate immune detection of bacterial virulence factors via the NLRC4 inflammasome. J Clin Immunol. 2010;30(4):502–6.CrossRefPubMedPubMedCentral

15.

Miao EA, Mao DP, Yudkovsky N, Bonneau R, Lorang CG, Warren SE, et al. Innate immune detection of the type III secretion apparatus through the NLRC4 inflammasome. Proc Natl Acad Sci U S A. 2010;107(7):3076–80.CrossRefPubMedPubMedCentral

16.

Miao EA, Rajan JV. Salmonella and caspase-1: a complex interplay of detection and evasion. Front Microbiol. 2011;2:85.CrossRefPubMedPubMedCentral

17.

Warren SE, Duong H, Mao DP, Armstrong A, Rajan J, Miao EA, et al. Generation of a Listeria vaccine strain by enhanced caspase-1 activation. Eur J Immunol. 2011;41(7):1934–40.CrossRefPubMedPubMedCentral

18.

Kupz A, Guarda G, Gebhardt T, Sander LE, Short KR, Diavatopoulos DA, et al. NLRC4 inflammasomes in dendritic cells regulate noncognate effector function by memory CD8(+) T cells. Nat Immunol. 2012;13(2):162–9.CrossRefPubMed

19.

Xu X, Hensel M. Systematic analysis of the SsrAB virulon of Salmonella enterica. Infect Immun. 2010;78(1):49–58.CrossRefPubMed

20.

Panthel K, Meinel KM, Domenech VES, Retzbach H, Igwe EI, Hardt WD, et al. Salmonella pathogenicity island 2-mediated overexpression of chimeric SspH2 proteins for simultaneous induction of antigen-specific CD4 and CD8 T Cells. Infect Immun. 2005;73(1):334–41.CrossRefPubMedPubMedCentral

21.

Medina C, Camacho EM, Flores A, Mesa-Pereira B, Santero E. Improved expression systems for regulated expression in Salmonella infecting eukaryotic cells. PLoS One. 2011;6(8):e23055.CrossRefPubMedPubMedCentral

22.

Meng FP, Ding J, Yu ZC, Han QL, Guo CC, Liu N, et al. Oral attenuated Salmonella typhimurium vaccine against MG7-Ag mimotope of gastric cancer. World J Gastroenterol. 2005;11(12):1833–6.CrossRefPubMedPubMedCentral

23.

Xu XG, Zhao HN, Zhang Q, Ding L, Li ZC, Li W, et al. Oral vaccination with attenuated Salmonella enterica serovar Typhimurium expressing Cap protein of PCV2 and its immunogenicity in mouse and swine models. Vet Microbiol. 2012;157:294–303.CrossRefPubMed

24.

Pelegrin P, Barroso-Gutierrez C, Surprenant A. P2X7 receptor differentially couples to distinct release pathways for IL-1β in mouse macrophage. J Immunol. 2008;180:7147–57.CrossRefPubMed

25.

Hoffmann C, Galle M, Dilling S, Kappeli R, Muller AJ, Songhet P, et al. In macrophages, caspase-1 activation by SopE and the type III secretion system-1 of S. Typhimurium can proceed in the absence of flagellin. PLoS One. 2010;5(8):e12477.CrossRefPubMedPubMedCentral

26.

Lightfield KL, Persson J, Trinidad NJ, Brubaker SW, Kofoed EM, Sauer JD, et al. Differential requirements for NAIP5 in activation of the NLRC4 inflammasome. Infect Immun. 2011;79(4):1606–14.CrossRefPubMedPubMedCentral

27.

Becker CE, O’Neill LAJ. Inflammasomes in inflammatory disorders: the role of TLRs and their interactions with NLRs. Semin Immunopathol. 2007;29:239–48.CrossRefPubMed

28.

Shaw MH, Reimer T, Kim YG, Nunez G. NOD-like receptors (NLRs): bona fide intracellular microbial sensors. Curr Opin Immunol. 2008;20(4):377–82.CrossRefPubMedPubMedCentral

29.

Byrne BG, Dubuisson JF, Joshi AD, Persson JJ, Swanson MS. Inflammasome components coordinate autophagy and pyroptosis as macrophage responses to infection. MBio. 2013;4(1):e00620–12.CrossRefPubMedPubMedCentral

30.

Figueira R, Holden DW. Functions of the Salmonella pathogenicity island 2 (SPI-2) type III secretion system effectors. Microbiology. 2012;158(Pt5):1147–61.CrossRefPubMed

31.

Pereira MSF, Marques GG, Deilama JE, Zamboni DS. The Nlrc4 inflammasome contributes to restriction of pulmonary infection by flagellated Legionella spp. that trigger pyroptosis. Front Microbiol. 2011;2:33.PubMedPubMedCentral

32.

Marlovits TC, Kubori T, Sukhan A, Thomas DR, Galan JE, Unger VM. Structural insights into the assembly of the type III secretion needle complex. Science. 2004;306(5698):1040–2.CrossRefPubMedPubMedCentral

33.

Yang X, Thornburg T, Suo Z, Jun S, Robison A, Li J, et al. Flagella overexpression attenuates Salmonella pathogenesis. PLoS One. 2012;7(10):e46828.CrossRefPubMedPubMedCentral

34.

Jones-Carson J, McCollister BD, Clambey ET, Vazquez-Torres A. Systemic CD8 T-cell memory response to a Salmonella pathogenicity island 2 effector is restricted to Salmonella enterica encountered in the gastrointestinal mucosa. Infect Immun. 2007;75(6):2708–16.CrossRefPubMedPubMedCentral

35.

Miao EA, Miller SI. A conserved amino acid sequence directing intracellular type III secretion by Salmonella typhimurium. Proc Natl Acad Sci U S A. 2000;97(13):7539–44.CrossRefPubMedPubMedCentral

36.

Lefebre MD, Galan JE. The inner rod protein controls substrate switching and needle length in a Salmonella type III secretion system. Proc Natl Acad Sci U S A. 2014;111(2):817–22.CrossRefPubMed

37.

Sal-Man N, Deng W, Finlay BB. EscI: a crucial component of the type III secretion system forms the inner rod structure in enteropathogenic Escherichia coli. Biochem J. 2012;442(1):119–25.CrossRefPubMed

38.

Zhao Z, Li M, Luo D, Xing L, Wu S, Duan Y, et al. Protection of mice from Brucella infection by immunization with attenuated Salmonella enterica serovar typhimurium expressing A L7/L12 and BLS fusion antigen of Brucella. Vaccine. 2009;27:5214–9.CrossRefPubMed

39.

Kong W, Brovold M, Koeneman BA, Clark-Curtiss J, Curtiss 3rd R. Turning self-destructing Salmonella into a universal DNA vaccine delivery platform. Proc Natl Acad Sci U S A. 2012;109(47):19414–9.CrossRefPubMedPubMedCentral

40.

Aachoui Y, Sagulenko V, Miao EA, Stacey KJ. Inflammasome-mediated pyroptotic and apoptotic cell death, and defense against infection. Curr Opin Microbiol. 2013;16(3):319–26.CrossRefPubMedPubMedCentral

41.

Martinon F, Burns K, Tschopp J. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-1β. Mol Cell. 2002;10:417–26.CrossRefPubMed

Title: Recombinant Salmonella expressing SspH2-EscI fusion protein limits its colonization in mice
Authors: Maozhi Hu
Weixin Zhao
Wei Gao
Wenhua Li
Chuang Meng
Qiuxiang Yan
Yuyang Wang
Xiaohui Zhou
Shizhong Geng
Zhiming Pan
Guiyou Cui
Xinan Jiao
Publication date: 01-12-2017
Publisher: BioMed Central
Published in: BMC Immunology / Issue 1/2017
Electronic ISSN: 1471-2172
DOI: https://doi.org/10.1186/s12865-017-0203-2

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Recombinant Salmonella expressing SspH2-EscI fusion protein limits its colonization in mice

Abstract

Background

Results

Conclusion

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2017

The immunoregulatory role of alpha enolase in dendritic cell function during Chlamydia infection

Giardia-specific cellular immune responses in post-giardiasis chronic fatigue syndrome

IG and TR single chain fragment variable (scFv) sequence analysis: a new advanced functionality of IMGT/V-QUEST and IMGT/HighV-QUEST

Autoantibodies against BAFF, APRIL or IL21 - an alternative pathogenesis for antibody-deficiencies?

Antigen presenting capacity of murine splenic myeloid cells

The imbalance of Th17/Treg cells is involved in the progression of nonalcoholic fatty liver disease in mice

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page