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. 2017 Oct 17;7:13367. doi: 10.1038/s41598-017-13570-2

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© The Author(s) 2017

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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Interferon-γ (IFN-γ) secretion by splenocytes from C57BL/6 J mice (n = 4/group) 49 days after vaccination with IB in combination with commercially available adjuvants (QuilA or Alum), nanoparticles (HEC, LRD or LFN) or PBS alone (control). Splenocytes (1.0 × 10⁵ cells/well) were harvested and co-cultured with IB (specific antigen), OVA (irrelevant antigen) or PBS (no antigen) for 40 h, then secretion of interferon-γ (IFN-γ) determined by ELISPOT assay. Photomicrographs show IFN-γ expressing splenocytes in response to (a) three hectorites and (b) best clay nanoparticle or commercial adjuvants; bar charts (c) and (d) illustrate the number of IFN-γ secreting spots in each vaccination group after stimulation with antigen (IB or OVA) or PBS. Data are expressed as mean ± S.E.M. (n = 4). *P < 0.05; **P < 0.01; ***P < 0.001; and ****P < 0.0001.