Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Dec 5;8(3):1542917. doi: 10.1080/2162402X.2018.1542917

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2018 The Author(s). Published with license by Taylor & Francis Group, LLC

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.

PMC Copyright notice

Figure 2. — Temporal induction of IL-12 in TCR-engineered T cells increases the numbers of T cells in the tumor and prevents PD-1 upregulation.

(a) Experimental setup. C57BL/6 female mice (Thy1.2⁺) were inoculated subcutaneously with 5 × 10⁵ B16 melanoma cells. 10 days later, mice were sublethally irradiated with 4Gy TBI 3-4hrs prior receiving intravenous injection of 2 × 10⁶ T cells (Thy1.1⁺) that were transduced with TRP2-TCR (T_TCR), Tet-IL-12 (T_iIL-12), TRP2-TCR + Tet-IL-12 (T_TCR+iIL-12), or mock-transduced T cells (T_mock). All mice received Dox (2mg/ml) in drinking water 2–3 days prior receiving T cell infusion and kept on Dox water for another 3 days. Tumors, spleens and lymph nodes were harvested on day 5 after the adoptive transfer and analyzed by flow cytometry. (b) Representative dot plots showing the percentage of transferred cells (Thy1.1⁺) (top) and pooled summary data (bottom). Cells were pre-gated on live-singlet lymphocytes. Symbols represent individual mice and bars indicate group averages. P values: T_TCR versus T_TCR+iIL-12 transduced cells and T_iIL-12 versus T_TCR+iIL-12 transduced cells; p < 0.0001. (c) Relative accumulation of TCR-expressing cells (CD19⁺) in tumor, spleen and draining lymph nodes (DLN) 5 days post T cell transfer. Cells were pre-gated on live-singlet transferred Thy1.1⁺ T cells. Data shown are cumulative results from at least two independent experiments. P values for the frequency of TCR⁺ cells: T_TCR versus pre-injection in the tumor, spleen and DLN; p > 0.05, T_TCR+iIL-12 versus pre-injection in the tumor; p = 0.0010, spleen; p = 0.0020 and DLN; p = 0.0049. P values for CD19 MFI: T_TCR versus T_TCR+iIL-12 in the tumor; p < 0.0001, spleen and DLN; p > 0.05. Symbols represent individual mice (T_TCR, n = 8 and T_TCR+iIL-12, n = 11). (d) Representative flow cytometry plots depicting the percentage of TCR-expressing cells (CD19⁺) and mean fluorescence intensity (MFI) of CD19 within the adoptively transferred Thy1.1⁺ T cells. Cells were pre-gated on live-singlet lymphocytes. (e) Representative flow cytometry plots depicting PD-1 staining profile of TCR-expressing cells (Vβ3⁺) in the tumor which divided them into two populations: Vβ3^hi PD-1^lo and Vβ3^lo PD-1^hi T cells (top) and bar charts representing summary data pooled from two independent experiments (bottom). P values: percentage of Vβ3^hi PD-1^lo T cells in T_TCR versus T_TCR+iIL-12; p < 0.0001, percentage of Vβ3^lo PD-1^hi T cells in T_TCR versus T_TCR+iIL-12; p = 0.0011.