In situ vaccination with defined factors overcomes T cell exhaustion in distant tumors
Danny N. Khalil, … , Jedd D. Wolchok, Taha Merghoub
Danny N. Khalil, … , Jedd D. Wolchok, Taha Merghoub
Published July 22, 2019
Citation Information: J Clin Invest. 2019;129(8):3435-3447. https://doi.org/10.1172/JCI128562.
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Research Article Oncology

In situ vaccination with defined factors overcomes T cell exhaustion in distant tumors

Abstract

Irreversible T cell exhaustion limits the efficacy of programmed cell death 1 (PD-1) blockade. We observed that dual CD40-TLR4 stimulation within a single tumor restored PD-1 sensitivity and that this regimen triggered a systemic tumor-specific CD8+ T cell response. This approach effectively treated established tumors in diverse syngeneic cancer models, and the systemic effect was dependent on the injected tumor, indicating that treated tumors were converted into necessary components of this therapy. Strikingly, this approach was associated with the absence of exhausted PD-1hi T cells in treated and distant tumors, while sparing the intervening draining lymph node and spleen. Furthermore, patients with transcription changes like those induced by this therapy experienced improved progression-free survival with anti–PD-1 treatment. Dual CD40-TLR4 activation within a single tumor is thus an approach for overcoming resistance to PD-1 blockade that is unique in its ability to cause the loss of exhausted T cells within tumors while sparing nonmalignant tissues.

Authors

Danny N. Khalil, Nathan Suek, Luis Felipe Campesato, Sadna Budhu, David Redmond, Robert M. Samstein, Chirag Krishna, Katherine S. Panageas, Marinela Capanu, Sean Houghton, Daniel Hirschhorn, Roberta Zappasodi, Rachel Giese, Billel Gasmi, Michael Schneider, Aditi Gupta, James J. Harding, John Alec Moral, Vinod P. Balachandran, Jedd D. Wolchok, Taha Merghoub

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Figure 2

CMP combination therapy augments APC activation and nodal accumulation followed by a systemic CD8+ T cell response.

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CMP combination therapy augments APC activation and nodal accumulation f...
(A) Using the bilateral tumor model, distant tumors from isotype- (Control) and CMP-treated (Trx) animals were assessed by H&E staining after 1 week of treatment (scale bars: 50 μm) and by flow cytometry to quantify CD8+ T cell infiltrates and the fraction of this cell population expressing Ki67 (n = 4/group). (B) Distant tumors were analyzed by immunofluorescence (IF) at 3 and 6 weeks for CD4 (green), FoxP3 (yellow), and CD8 (red) cell populations (scale bars: 50 μm). Quantification of the CD8+ fraction of DAPI+ cells in IF images (n = 3–10/group). N/A, no remaining live animals. (C) Growth of treated and distant tumors from WT or Rag1–/– C57BL/6 animals (n = 10/group). (D) Growth of treated and distant tumors from mice depleted of CD4+ and CD8+ T cells. Peripheral blood was collected to confirm the absence of corresponding cell populations (n = 10/group). (E) Tumor growth in mice bearing treated WT B16F10 and distant B78H10 tumors (n = 9–10/group). (F) Mice previously cured of unilateral B16F10 tumors with CMP treatment and age-matched naive controls were implanted with tumors on day 90 (n = 8–10/group). Adjacent panel shows fur depigmentation at the site of the initial cured tumor (green arrowhead) and at the site of post-treatment tumor reimplantation (red arrowhead). (G) CD86 expression in the CD11chi cell population in the treated tumor and DLN (n = 4/group). (H) Fraction of CD11chi cells among live CD45+ cells in the treated tumor and DLN (n = 4/group). (I) Tumor growth of WT or Batf3–/– C57BL/6 animals bearing B16F10 tumors treated with isotype or CMP (n = 10/group). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001, by unpaired, 2-tailed Student’s t test.