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. 2016 Mar 10;8(3):78. doi: 10.3390/v8030078

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© 2016 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).

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Frequencies of double positive phenotypes of ASP-specific T-cell responses in immunized mice. (A) Study design: C57BL/6 mice were immunized intramuscularly with Ad5, Ad5-CMV-ASP-2, Ad48, or Ad48-CMV-ASP-2 at the indicated time points. Two weeks after the last immunization, splenocytes harvested from a subset of mice were stimulated with anti-CD107a and GolgiStop in the presence or absence of ASP-C peptide. Cells were gated by side scatter area (SSC-A); forward scatter area (FSC-A); and gated for CD3⁺ CD4⁺ or CD3⁺ CD8⁺ T lymphocytes; (B) Representative flow cytometry plots of CD107a⁺ and IFNγ⁺ by CD8⁺ T-cells from the spleens of the immunized mice; (C) Percentages of CD4⁺ T-cells which produce CD107a⁺IFNγ⁺ or CD107a⁺TNFα⁺ after stimulation with ASP-C peptide; (D) Percentages of CD8⁺ T-cells which produce CD107a⁺IFNγ⁺ or CD107a⁺TNFα⁺ after stimulation with ASP-C peptide. Results are presented as the mean ± SEM frequencies of CD4⁺ or CD8⁺ cells for 4 mice. (*) = p ≤ 0.05.

Frequencies of double positive phenotypes of ASP-specific T-cell responses in immunized mice. (A) Study design: C57BL/6 mice were immunized intramuscularly with Ad5, Ad5-CMV-ASP-2, Ad48, or Ad48-CMV-ASP-2 at the indicated time points. Two weeks after the last immunization, splenocytes harvested from a subset of mice were stimulated with anti-CD107a and GolgiStop in the presence or absence of ASP-C peptide. Cells were gated by side scatter area (SSC-A); forward scatter area (FSC-A); and gated for CD3⁺ CD4⁺ or CD3⁺ CD8⁺ T lymphocytes; (B) Representative flow cytometry plots of CD107a⁺ and IFNγ⁺ by CD8⁺ T-cells from the spleens of the immunized mice; (C) Percentages of CD4⁺ T-cells which produce CD107a⁺IFNγ⁺ or CD107a⁺TNFα⁺ after stimulation with ASP-C peptide; (D) Percentages of CD8⁺ T-cells which produce CD107a⁺IFNγ⁺ or CD107a⁺TNFα⁺ after stimulation with ASP-C peptide. Results are presented as the mean ± SEM frequencies of CD4⁺ or CD8⁺ cells for 4 mice. (*) = p ≤ 0.05.