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. 2016 Oct 14;6:35173. doi: 10.1038/srep35173

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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(a) OT-II transferred mice were immunized with aAVC-OVA or Alum plus OVA protein (100 μg). Fourteen days later, germinal center (GC) formation in spleen was analyzed by flow cytometric analysis using B220-APC, CD95-PE and GL7-FITC. (Mean ± SEM, n = 4–6.) ***P < 0.001. (b) GC formation in spleen sections was verified by confocal microscopy using IgD-PE and PNA-Alexa 647. Scale bar, 200 μm (c) ELISPOT analysis of OVA-specific IgG-secreting splenic B cells. (Mean ± SEM, n = 4). *P < 0.05. (d) Localization of OT-II cells in OT-II transferred mice immunized with aAVC-OVA (left panel) or non-immunized (right panel) was analyzed using CD45.1-FITC, B220-APC, and CD169-PE 7 days after immunization or 3 days after a transfer of OT-II cells. The dotted line shows approximate location of the boundary between the B cell follicle and the T cell zone. All data are representative of at least four experiments. Scale bar, 50 μm.