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. 2017 Jun 23;12:4579–4591. doi: 10.2147/IJN.S131309

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© 2017 Di Bonito et al. This work is published and licensed by Dove Medical Press Limited

The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed.

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Antitumor therapeutic effect induced by IM inoculation of Nef^mut/E7 DNA vector.

Notes: C57 Bl/6 mice were challenged with 2×10⁵ TC-1 cells, and 4 days later (arrows), when tumor masses became detectable by palpation, inoculated with either Nef^mut/E7 vector (seven mice) Nef^mut vector, empty vector, or vehicle (four mice per group). The DNA inoculations were repeated at day 11 (arrows) after tumor cell implantation, and the growth of tumor mass was followed over time. (A) E7-specific CD8⁺ T-cell response as detected by IFN-γ Elispot assay was carried out with PBMCs recovered from retro-orbital bleeding 7 days after the last immunization and cultivated for 16 hours in the presence of either unrelated or E7 peptides. As control, PBMCs were incubated with 5 ng/mL of PMA and 500 ng/mL of ionomycin. Shown are the number of SFU/10⁵ cells from triplicate wells seeded with splenocytes from mice inoculated with either Nef^mut or Nef^mut/E7 vectors. (B) Determination of the tumor size during the 30-day observation time. (C) Weight of tumors from mice injected with either Nef^mut or Nef^mut/E7 DNA vectors at the time of sacrifice. Shown are the results of one representative of two independent experiments.

Abbreviations: IM, intramuscular; PBMCs, peripheral blood mononuclear cells; PMA, phorbol 12-myristate 13-acetate; SFU, spot-forming unit.

Antitumor therapeutic effect induced by IM inoculation of Nef^mut/E7 DNA vector.

Notes: C57 Bl/6 mice were challenged with 2×10⁵ TC-1 cells, and 4 days later (arrows), when tumor masses became detectable by palpation, inoculated with either Nef^mut/E7 vector (seven mice) Nef^mut vector, empty vector, or vehicle (four mice per group). The DNA inoculations were repeated at day 11 (arrows) after tumor cell implantation, and the growth of tumor mass was followed over time. (A) E7-specific CD8⁺ T-cell response as detected by IFN-γ Elispot assay was carried out with PBMCs recovered from retro-orbital bleeding 7 days after the last immunization and cultivated for 16 hours in the presence of either unrelated or E7 peptides. As control, PBMCs were incubated with 5 ng/mL of PMA and 500 ng/mL of ionomycin. Shown are the number of SFU/10⁵ cells from triplicate wells seeded with splenocytes from mice inoculated with either Nef^mut or Nef^mut/E7 vectors. (B) Determination of the tumor size during the 30-day observation time. (C) Weight of tumors from mice injected with either Nef^mut or Nef^mut/E7 DNA vectors at the time of sacrifice. Shown are the results of one representative of two independent experiments.

Abbreviations: IM, intramuscular; PBMCs, peripheral blood mononuclear cells; PMA, phorbol 12-myristate 13-acetate; SFU, spot-forming unit.