Memory T cell–driven differentiation of naive cells impairs adoptive immunotherapy
Christopher A. Klebanoff, … , Richard M. Siegel, Nicholas P. Restifo
Christopher A. Klebanoff, … , Richard M. Siegel, Nicholas P. Restifo
Published December 14, 2015
Citation Information: J Clin Invest. 2016;126(1):318-334. https://doi.org/10.1172/JCI81217.
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Research Article Immunology Oncology Article has an altmetric score of 28

Memory T cell–driven differentiation of naive cells impairs adoptive immunotherapy

Abstract

Adoptive cell transfer (ACT) of purified naive, stem cell memory, and central memory T cell subsets results in superior persistence and antitumor immunity compared with ACT of populations containing more-differentiated effector memory and effector T cells. Despite a clear advantage of the less-differentiated populations, the majority of ACT trials utilize unfractionated T cell subsets. Here, we have challenged the notion that the mere presence of less-differentiated T cells in starting populations used to generate therapeutic T cells is sufficient to convey their desirable attributes. Using both mouse and human cells, we identified a T cell–T cell interaction whereby antigen-experienced subsets directly promote the phenotypic, functional, and metabolic differentiation of naive T cells. This process led to the loss of less-differentiated T cell subsets and resulted in impaired cellular persistence and tumor regression in mouse models following ACT. The T memory–induced conversion of naive T cells was mediated by a nonapoptotic Fas signal, resulting in Akt-driven cellular differentiation. Thus, induction of Fas signaling enhanced T cell differentiation and impaired antitumor immunity, while Fas signaling blockade preserved the antitumor efficacy of naive cells within mixed populations. These findings reveal that T cell subsets can synchronize their differentiation state in a process similar to quorum sensing in unicellular organisms and suggest that disruption of this quorum-like behavior among T cells has potential to enhance T cell–based immunotherapies.

Authors

Christopher A. Klebanoff, Christopher D. Scott, Anthony J. Leonardi, Tori N. Yamamoto, Anthony C. Cruz, Claudia Ouyang, Madhu Ramaswamy, Rahul Roychoudhuri, Yun Ji, Robert L. Eil, Madhusudhanan Sukumar, Joseph G. Crompton, Douglas C. Palmer, Zachary A. Borman, David Clever, Stacy K. Thomas, Shashankkumar Patel, Zhiya Yu, Pawel Muranski, Hui Liu, Ena Wang, Francesco M. Marincola, Alena Gros, Luca Gattinoni, Steven A. Rosenberg, Richard M. Siegel, Nicholas P. Restifo

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Figure 7

Precocious differentiation is associated with augmented Akt signaling.

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Precocious differentiation is associated with augmented Akt signaling. R...
Representative FACS histograms and summary bar graphs demonstrating the mean fluorescence intensity of (A) pAkt T308, (B) pAkt S473, and (C) pS6 in Ly5.2+CD8+ TN cells at rest or 24 hours after stimulation with CD3/CD28-specific antibodies alone or in a 1:1 mixture with Ly5.1+ TMem cells. Data shown after gating on live+Ly5.2+CD8+ cells. (D) RMA-normalized intensity showing expression of Il7ra, S1pr1, Tfrc, Hk2, and Slc2a1 in resting TN cells, TN cells primed alone, and TN cells primed in a 1:1 mixture with TMem cells. (E) Western blot and densitometry analysis of pAkt T308 and GAPDH in resting TN or TN cells primed for 24 hours with CD3/CD28-specific antibodies and titrated doses of lz-FasL. (F) Representative FACS plot and (G) summary scatter plot showing 2-NBDG expression in TN cells primed alone for 6 days with CD3/CD28-specific antibodies, IL-2, and lz-FasL (50 ng/ml) or vehicle control. Data shown after gating on live+CD8+ lymphocytes. (H) Representative FACS plots, (I) summary bar graph demonstrating the frequency of CD8+ T cell subsets, and (J) scatter plots showing IFN-γ production in TN cells primed with CD3/CD28-specific antibodies, IL-2, and lz-FasL (50 ng/ml) in the presence or absence of an inhibitor of Akt1/2 (AktI) for 6 days. Statistical comparisons performed using an unpaired 2-tailed Student’s t test corrected for multiple comparisons by a Bonferroni adjustment. *P < 0.05; **P < 0.01; ***P < 0.001. All data shown are displayed as mean ± SEM. Experiments performed with n = 3 per condition/time point in A–D and F–J. All data shown are representative of 2 independently conducted experiments.