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. 2019 Dec 10;9:18713. doi: 10.1038/s41598-019-55014-z

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© The Author(s) 2019

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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5GHPV3-specific CD8 + T cells with an effector phenotype are detected in the cervix following administration of heterologous prime-boost vaccine regimens. (A) C57BL/6 mice (6/group) were vaccinated with DM or CM regimens. Cervicovaginal lymphocytes were isolated six weeks post-boost, stimulated with 5GHPV3 E6 and E7 peptides and analysed by ICS for IFN-ɣ, CD107, TNF-α and IL-2. (B) For CM-vaccinated mice, cervicovaginal lymphocyte responses were compared at one and six weeks post-boost. Mice were pooled due to low cell numbers. (C) Comparison of CD49d/CD29 (α4β1) expression in 5GHPV3 E6-specific T cells among cervicovaginal lymphocytes, splenocytes and PBMCs at two weeks post-boost in CM-vaccinated C57BL/6 mice and (D) in CM vs. DM vaccinated C57BL/6 mice. Expression of differentiation/memory markers in 5GHPV3 E6 or E7-specific T cells was assessed in (E) cervicovaginal lymphocytes and (F) splenocytes from C57BL/6 mice (6/group) vaccinated with CM at one week (top panels) and three weeks (bottom panels) post-boost. Samples were pooled due to low cell number. Effector (CD62L⁻, CD127⁻), effector memory (CD62L⁻, CD127+) and central memory (CD62+, CD127+) populations are shown. Horizontal lines show mean with standard deviation. Two-way ANOVA, Sidak’s multiple comparison test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001.