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. 2016 Aug 12;6:82. doi: 10.3389/fcimb.2016.00082

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Copyright © 2016 Kuo, Chen, Chiu, Teng, Hsu, Lu, Syu, Wang, Chou and Chen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Deletion of EHEC rfaD increases its susceptibility to human serum killing and the antimicrobial peptide LL-37. The fold changes of bacterial cell numbers of wild-type EDL933, EDL933 rfaD::Tn5, EDL933:ΔrfaD and EDL933:ΔrfaD-prfaD after incubation in 10% normal human serum (NHS) (A) or 10% heat-inactivated normal human serum (HNHS) (B) at 37°C for 60 min were measured. Experiments were conducted independently at least three times, and error bars indicate the standard deviations. (A) The rfaD transposon mutant (rfaD::Tn5, P = 0.001) and rfaD deletion mutant (ΔrfaD, P = 0.001) showed significant decreased survival ratio compared to that of wild-type EDL933 (EDL933) in 10% NHS. The rfaD complement bacteria (ΔrfaD-prfaD) showed comparable survival ratio compared to that of EDL933 (P = 0.101) in 10% NHS. (B) The rfaD transposon mutant (rfaD::Tn5, P = 0.031) and rfaD deletion mutant (ΔrfaD, P = 0.034) showed significantly decreased survival ratio compared to that of wild-type EDL933 (EDL933) in 10% HNHS. The rfaD complement bacteria (ΔrfaD-prfaD) showed comparable survival ratio compared to that of EDL933 (P = 0.854) in 10% HNHS. n.s. indicates no statistical significance. (C) The OD₅₉₅ values of wild-type EDL933 (EDL933), EDL933 rfaD::Tn5 (rfaD::Tn5), EDL933:ΔrfaD (ΔrfaD), and EDL933:ΔrfaD-prfaD (ΔrfaD-prfaD) cultured with different dose of LL-37 at 37°C for 16 h were monitored. (D) The growth curves of EDL933, rfaD::Tn5, ΔrfaD and ΔrfaD-prfaD in the presence of 12.5 μg/ml LL-37 at 37°C.