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. 2018 Feb 12;8:2786. doi: 10.1038/s41598-018-21134-1

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Immune response to intravaginal application of C. crescentus. (a) PBS, 20 μg LPS from Salmonella enterica serotype Minnesota or 10⁸ Cc-Control were applied to the vaginal tract of progesterone treated C57Bl/6 mice. Vaginal lavage fluid was collected prior to application then 4, 24 and 48 hours post-application and analyzed by cytometric bead array for mouse inflammatory cytokines. Statistical analysis was performed by two-way ANOVA with Bonferroni’s correction for multiple comparisons. *p ≤ 0.05, **p ≤ 0.01. n = 4–5 (TNF), n = 7–11 (remaining cytokines). (b) Hematoxylin and eosin stain vaginal lavage tissue 4 hours after application of PBS, LPS from Salmonella enterica serotype Minnesota or C. crescentus. (c) Mice were inoculated intravaginally with 10⁸ C. crescentus biweekly for 9 weeks before vaginal lavage fluid was collected and analyzed by ELISA for antibodies against C. crescentus. An anti-RsaA antibody raised in rabbits was used to create a standard curve (limit of detection 14 pg/mL) and values were interpolated from the standard curve.