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. 2020 Nov 6;8(2):e001355. doi: 10.1136/jitc-2020-001355

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© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.

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Proof-in-concept that inhaled IL-15 drives peripheral NK and CD8⁺ T cell activation and TIGIT upregulation in dogs with metastatic osteosarcoma. (A) Schema for our first-in-dog clinical trial of inhaled IL-15 in dogs with naturally occurring metastatic osteosarcoma. (B) Representative flow cytometry showing peripheral blood CD45⁺, lymphocytes, CD3, CD3⁻NKp46⁺ NK, and CD3⁺CD8⁺ T cell populations. (C–D) Inhaled IL-15 stimulates peripheral cytotoxic lymphocytes in vivo leading to upregulation of granzyme B and Ki67 by flow cytometry analysis. (E–F) Fold change in granzyme B expression significantly increased on NK cells (p=0.01) and fold change in Ki67 expression significantly increased on T cells (p=0.03). (G) RNA sequencing analysis of CD5-depleted NK cells from three dogs on-trial showing TIGIT upregulation in 2/3 patients. Mean±SEM. P values determined using paired Student’s t-test. Fold change comparison using Pre Tx value of 1. IL-15, interleukin 15; NK, natural killer; On Tx, on treatment; Pre Tx, pretreatment.