The online version of this article (https://doi.org/10.1186/s12974-018-1293-3) contains supplementary material, which is available to authorized users.
Listeria monocytogenes is a common cause of bacterial meningitis. We developed an animal model of listerial meningitis.
In survival studies, C57BL/6 mice received intracisternal injections with different L. monocytogenes sequence type 1 (ST1) colony forming units per milliliter (CFU; n = 48, 105, 106, 107, 108, and 109 CFU/ml). Second, mice were inoculated with 108 CFU/ml ST1 and sacrificed at 6 h and 24 h (n = 12/group). Outcome parameters were clinical score, CFUs, cyto- and chemokine levels, and brain histopathology. Third, 84 mice were inoculated (109 CFU/ml ST1) to determine optimal antibiotic treatment with different doses of amoxicillin and gentamicin. Fourth, mice were inoculated with 109 CFU/ml ST1, treated with amoxicillin, and sacrificed at 16 h and 24 h (n = 12/group) for outcome assessment. Finally, time point experiments were repeated with ST6 (n = 24/group).
Median survival time for inoculation with 108 and 109 CFU/ml ST1 was 46 h and 40 h; lower doses of bacteria led to minimal clinical signs of disease. Brain levels of IL-6, IL-17A, and IFN-γ were elevated at 24 h, and IL-1β, IL-6, IL-10, IFN-γ, and TNF-α were elevated in blood at 6 h and 24 h. Histopathology showed increased meningeal infiltration, vascular inflammation of meningeal vessels, hemorrhages, and ventriculitis. In the treatment model, brain levels of IL-6 and IL-17A and blood levels of IL-6 and IFN-γ were elevated. Compared to ST6, infection with ST1 led initially to higher levels of IL-1β and TNF-α in blood and more profound neuropathological damage. At 16 h post inoculation, IL-1β, IL-10, and TNF-α in blood and IL-6, IL17A, TNF-α, and IFN-γ levels in brain were higher in ST1 compared to ST6 without differences in CFUs between STs. At 24 h, neuropathology score was higher in ST1 compared to ST6 (p = 0.002) infected mice.
We developed and validated a murine model of listerial meningitis. ST1-infected mice had a more severe inflammatory response and brain damage as compared to ST6-infected mice.
Additional file 1: This table shows a histopathological scoring method of brain tissue in bacterial meningitis mouse model which has been used in this study and previously has been used in a pneumococcal meningitis model. (DOC 48 kb)12974_2018_1293_MOESM1_ESM.doc
Additional file 2: Kaplan-Meier survival curve (A) in male and female mice (24 mice/group). Clinical score of the treatment survival experiments (12 mice/ group) inoculated with 109 CFU bacteria and treated with antibiotics. Abbreviation; h = hours (PDF 30 kb)12974_2018_1293_MOESM2_ESM.pdf
Additional file 3: This table shows histopathological scoring of brain tissue in listerial meningitis time point studies with L. monocytogenes ST1 and ST6 strains. Results are presented based on number of mice and on median pathology score. (DOC 82 kb)12974_2018_1293_MOESM3_ESM.doc
Additional file 4: Kaplan-Meier survival curves in treatment survival experiments inoculated with 109 CFU/ml (A and B) and with 108 CFU/ml (C) and bacterial outgrowth after inoculation with 109 CFU/ml L. monocytogenes ST1 and amoxicillin treatment (D). --- lower limit of detection, Abbreviation; h = hours. (PDF 44 kb)12974_2018_1293_MOESM4_ESM.pdf
Additional file 5: This table shows bacterial outgrowth in brain homogenate in mice infected with L. monocytogenes ST1 and treated with antibiotics during survival experiments (70 h post inoculation). Every bacterial count represents one mouse. (DOC 51 kb)12974_2018_1293_MOESM5_ESM.doc
Additional file 6: (A) Median clinical score in ST1 and ST6 inoculated mice in the non-treatment model with interquartile ranges, (B) Bacterial outgrowth in the non-treatment model ST1 vs. ST6 6 h after inoculation. Titres are expressed per mice and with median CFU/ml or CFU/mg. (PDF 38 kb)12974_2018_1293_MOESM6_ESM.pdf
Additional file 7: This table shows the brain and plasma levels of cytokines in mice infected with 109 CFU/ml L. monocytogenes ST1 or ST6 at time points 16 and 24 h and treated with 100 mg/kg/24 h amoxicillin after 16 h. (DOC 64 kb)12974_2018_1293_MOESM7_ESM.doc
Charlier C, Perrodeau E, Leclercq A, Cazenave B, Pilmis B, Henry B, Lopes A, Maury MM, Moura A, Goffinet F, et al. Clinical features and prognostic factors of listeriosis: the MONALISA national prospective cohort study. Lancet Infect Dis. 2017;15:150–9.
Labow M, Shuster D, Zetterstrom M, Nunes P, Terry R, Cullinan EB, Bartfai T, Solorzano C, Moldawer LL, Chizzonite R, McIntyre KW. Absence of IL-1 signaling and reduced inflammatory response in IL-1 type I receptor-deficient mice. J Immunol. 1997;159:2452–61. PubMed
Tripp CS, Gately MK, Hakimi J, Ling P, Unanue ER. Neutralization of IL-12 decreases resistance to listeria in SCID and C.B-17 mice. Reversal by IFN-gamma. J Immunol. 1994;152:1883–7. PubMed
Tripp CS, Wolf SF, Unanue ER. Interleukin 12 and tumor necrosis factor alpha are costimulators of interferon gamma production by natural killer cells in severe combined immunodeficiency mice with listeriosis, and interleukin 10 is a physiologic antagonist. Proc Natl Acad Sci U S A. 1993;90:3725–9. CrossRefPubMedPubMedCentral
Bou Ghanem EN, Myers-Morales T, D'Orazio SE. A mouse model of foodborne listeria monocytogenes infection. Curr Protoc Microbiol. 2013;31:9B 3 1–9B 3 16.
Maury MM, Tsai YH, Charlier C, Touchon M, Chenal-Francisque V, Leclercq A, Criscuolo A, Gaultier C, Roussel S, Brisabois A, et al. Uncovering listeria monocytogenes hypervirulence by harnessing its biodiversity. Nat Genet. 2016;48:308–13.
Seebach J, Bartholdi D, Frei K, Spanaus KS, Ferrero E, Widmer U, Isenmann S, Strieter RM, Schwab M, Pfister H, Fontana A. Experimental listeria meningoencephalitis. Macrophage inflammatory protein-1 alpha and -2 are produced intrathecally and mediate chemotactic activity in cerebrospinal fluid of infected mice. J Immunol. 1995;155:4367–75. PubMed
Koopmans MM, Bijlsma MW, Brouwer MC, van de Beek D, van der Ende A. Listeria monocytogenes meningitis in the Netherlands, 1985-2014: a nationwide surveillance study. J Inf. 2017;75:12–9.
Becavin C, Bouchier C, Lechat P, Archambaud C, Creno S, Gouin E, Wu Z, Kuhbacher A, Brisse S, Pucciarelli MG, et al. Comparison of widely used listeria monocytogenes strains EGD, 10403S, and EGD-e highlights genomic variations underlying differences in pathogenicity. MBio. 2014;5:e00969–14. CrossRefPubMedPubMedCentral
Kowalik MM, Smiatacz T, Hlebowicz M, Pajuro R, Trocha H. Coagulation, coma, and outcome in bacterial meningitis--an observational study of 38 adult cases. J Inf Secur. 2007;55:141–8.
Weisfelt M, Determann RM, de Gans J, van der Ende A, Levi M, van de Beek D, Schultz MJ. Procoagulant and fibrinolytic activity in cerebrospinal fluid from adults with bacterial meningitis. J Inf Secur. 2007;54:545–50.
Havell EA, Sehgal PB. Tumor necrosis factor-independent IL-6 production during murine listeriosis. J Immunol. 1991;146:756–61. PubMed
Rothe J, Lesslauer W, Lotscher H, Lang Y, Koebel P, Kontgen F, Althage A, Zinkernagel R, Steinmetz M, Bluethmann H. Mice lacking the tumour necrosis factor receptor 1 are resistant to TNF-mediated toxicity but highly susceptible to infection by listeria monocytogenes. Nature. 1993;364:798–802. CrossRefPubMed
Pfeffer K, Matsuyama T, Kundig TM, Wakeham A, Kishihara K, Shahinian A, Wiegmann K, Ohashi PS, Kronke M, Mak TW. Mice deficient for the 55 kd tumor necrosis factor receptor are resistant to endotoxic shock, yet succumb to L. monocytogenes infection. Cell. 1993;73:457–67.
Dai WJ, Kohler G, Brombacher F. Both innate and acquired immunity to listeria monocytogenes infection are increased in IL-10-deficient mice. J Immunol. 1997;158:2259–67. PubMed
Kremer PH, Lees JA, Koopmans MM, Ferwerda B, Arends AW, Feller MM, Schipper K, Valls Seron M, van der Ende A, Brouwer MC, et al. Benzalkonium tolerance genes and outcome in listeria monocytogenes meningitis. Clin Microbiol Infect. 2017;23:265.e1-265.e7.
Ghosh P, Halvorsen EM, Ammendolia DA, Mor-Vaknin N, O'Riordan MXD, Brumell JH, Markovitz DM, Higgins DE. Invasion of the brain by listeria monocytogenes is mediated by InlF and host cell Vimentin. MBio. 2018;9
Lee S, Ward TJ, Jima DD, Parsons C, Kathariou S. The arsenic resistance-associated listeria genomic island LGI2 exhibits sequence and integration site diversity and a propensity for three listeria monocytogenes clones with enhanced virulence. Appl Environ Microbiol. 2017;83.
Kasanmoentalib ES, Valls Seron M, Ferwerda B, Tanck MW, Zwinderman AH, Baas F, van der Ende A, Brouwer MC, van de Beek D. Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitis. J Neuroinflammation. 2017;14:2. CrossRefPubMedPubMedCentral
Mook-Kanamori BB, Valls Seron M, Geldhoff M, Havik SR, van der Ende A, Baas F, van der Poll T, Meijers JC, B PM, Brouwer MC, van de Beek D. Thrombin-activatable fibrinolysis inhibitor influences disease severity in humans and mice with pneumococcal meningitis. J Thromb Haemost. 2015;13:2076–86. CrossRefPubMed
- Characterization of a Listeria monocytogenes meningitis mouse model
Merel M. Koopmans
Matthijs C. Brouwer
Wing Kit Man
Mercedes Vall Seron
Diederik van de Beek
- BioMed Central