Clinical Microbiology and Infection

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Clinical Microbiology and Infection Reassessing the role of internalin B in Listeria monocytogenes virulence using the epidemic strain F2365 J.J. Quereda, I.M. Rodríguez-Gómez, J. Meza-Torres, J. Gómez-Laguna, M.A. Nahori, O. Dussurget, L. Carrasco, P. Cossart, J. Pizarro- Cerdá Clinical Microbiology and Infection DOI: 10.1016/j.cmi.2018.08.022 Copyright © 2018 The Authors Terms and Conditions

Fig. 1 (A) InlB protein levels detected by Western blot analysis. Levels of loading control protein, EF-Tu, are shown for comparison. (B) Role of InlB in epidemic Listeria monocytogenes entry. Eukaryotic cells were incubated with bacteria for 30 minutes (JEG-3 and RAW 264.7) or 1 hour (HeLa) at 37 °C. After incubation, extracellular bacteria were neutralized by adding complete fresh medium containing 40 μg/mL of gentamicin. At 2 hours after infection, cells were lysed to determine number of viable intracellular L. monocytogenes. Bars show numbers of viable intracellular bacteria. Mean and standard deviation are shown. Three independent experiments with 6 replicates in each experiment were performed. Data from one representative experiment are shown. ****p <0.0001, **p <0.01; ns indicates not statistically significant by Student's t test. Error bars represent standard deviation. (C) BALB/c mice were intravenously inoculated with 104 L. monocytogenes F2365 and F2365 InlB+. Half of organ was used to assess bacteria load and other half was used for histopathologic analysis. CFUs in spleen and liver were assessed at 72 and 96 hours after infection. Each dot represents value for one mouse. Two mice infected with F2365 InlB+ strain died before 96 hours. Statistically significant differences were evaluated by Mann-Whitney test. *p <0.05. (D) Histopathologic analysis of number of necrotic foci and ratio of necrotic area to total area were recorded in spleens and livers from same infected mice (*p <0.05 by Mann-Whitney test). Clinical Microbiology and Infection DOI: (10.1016/j.cmi.2018.08.022) Copyright © 2018 The Authors Terms and Conditions