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. 2016 Dec 7;7:559. doi: 10.3389/fimmu.2016.00559

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Copyright © 2016 Guttman, Freixo-Lima, Kaner, Lior, Rider and Lewis.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Increased activation marker expression by RMA tumor-infiltrating leukocytes in hAAT-treated mice. (A) PBS- and hAAT-treated RMA tumor-bearing mice (n = 4 per group) were sacrificed 13 days after RMA cell inoculation (1 × 10⁵ cells per animal, s.c.). Tumors were dissociated, and cell populations were stained for indicated markers. (B) Tumors and spleens of RMA tumor-bearing animals were dissociated, and T cell populations were stained and analyzed by flow cytometry. (C,D) Purified splenic or tumor-infiltrating CD8⁺ T cells (1 × 10⁵ cells per well in triplicate) were incubated for 24 h with or without cultured RMA cells (CT:RMA, respectively) (1:1 ratio). Cells were then stained for (C) intracellular perforin and TNFα, and (D) release of TNFα to the supernatant was quantified. Mean ± SEM, *p < 0.05, **p < 0.01. Unpaired two-tailed Student’s t-test was employed to assess differences between groups.

Increased activation marker expression by RMA tumor-infiltrating leukocytes in hAAT-treated mice. (A) PBS- and hAAT-treated RMA tumor-bearing mice (n = 4 per group) were sacrificed 13 days after RMA cell inoculation (1 × 10⁵ cells per animal, s.c.). Tumors were dissociated, and cell populations were stained for indicated markers. (B) Tumors and spleens of RMA tumor-bearing animals were dissociated, and T cell populations were stained and analyzed by flow cytometry. (C,D) Purified splenic or tumor-infiltrating CD8⁺ T cells (1 × 10⁵ cells per well in triplicate) were incubated for 24 h with or without cultured RMA cells (CT:RMA, respectively) (1:1 ratio). Cells were then stained for (C) intracellular perforin and TNFα, and (D) release of TNFα to the supernatant was quantified. Mean ± SEM, *p < 0.05, **p < 0.01. Unpaired two-tailed Student’s t-test was employed to assess differences between groups.