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. 2015 May;125(5):2046-58.
doi: 10.1172/JCI80445. Epub 2015 Apr 13.

TIGIT and PD-1 impair tumor antigen-specific CD8⁺ T cells in melanoma patients

Joe-Marc Chauvin, Ornella Pagliano, Julien Fourcade, Zhaojun Sun, Hong Wang, Cindy Sander, John M Kirkwood, Tseng-hui Timothy Chen, Mark Maurer, Alan J Korman, Hassane M Zarour

TIGIT and PD-1 impair tumor antigen-specific CD8⁺ T cells in melanoma patients

Joe-Marc Chauvin et al. J Clin Invest. 2015 May.

Abstract

T cell Ig and ITIM domain (TIGIT) is an inhibitory receptor expressed by activated T cells, Tregs, and NK cells. Here, we determined that TIGIT is upregulated on tumor antigen-specific (TA-specific) CD8⁺ T cells and CD8⁺ tumor-infiltrating lymphocytes (TILs) from patients with melanoma, and these TIGIT-expressing CD8⁺ T cells often coexpress the inhibitory receptor PD-1. Moreover, CD8⁺ TILs from patients exhibited downregulation of the costimulatory molecule CD226, which competes with TIGIT for the same ligand, supporting a TIGIT/CD226 imbalance in metastatic melanoma. TIGIT marked early T cell activation and was further upregulated by T cells upon PD-1 blockade and in dysfunctional PD-1⁺TIM-3⁺ TA-specific CD8⁺ T cells. PD-1⁺TIGIT⁺, PD-1⁻TIGIT⁺, and PD-1⁺TIGIT⁻ CD8⁺ TILs had similar functional capacities ex vivo, suggesting that TIGIT alone, or together with PD-1, is not indicative of T cell dysfunction. However, in the presence of TIGIT ligand-expressing cells, TIGIT and PD-1 blockade additively increased proliferation, cytokine production, and degranulation of both TA-specific CD8⁺ T cells and CD8⁺ TILs. Collectively, our results show that TIGIT and PD-1 regulate the expansion and function of TA-specific CD8⁺ T cells and CD8⁺ TILs in melanoma patients and suggest that dual TIGIT and PD-1 blockade should be further explored to elicit potent antitumor CD8⁺ T cell responses in patients with advanced melanoma.

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Figures

Figure 8
Figure 8. CD8+ TILs downregulate CD226 expression in metastatic melanoma.
(A and B) Pooled data comparing the frequencies (A) and MFI (B) of CD226 expression by CD8+ T cells isolated from PBMCs from healthy donors (n = 6) and melanoma patients (n = 6), by circulating NY-ESO-1–specific CD8+ T cells from melanoma patients (n = 6), and by CD8+ TILs (n = 7) isolated from metastatic melanoma. **P < 0.01 and ***P < 0.001 by Kruskal-Wallis test, followed by Dunn’s multiple comparisons test. Data are representative of 3 independent experiments.
Figure 7
Figure 7. CD8+ TILs upregulate TIGIT and PD-1 and exhibit enhanced proliferation and degranulation upon dual TIGIT and PD-1 blockade.
(A) Pooled data showing TIGIT expression (percentage and MFI) by CD8+ T cells from PBMCs from healthy donors (HD) (n = 8), melanoma patients (MP) (n = 13), and melanoma TILs (TILs) (n = 18). P values were obtained by a 1-way ANOVA, followed by Tukey’s multiple comparisons test (left panel) and a Kruskal-Wallis test, followed by Dunn’s multiple comparisons test (right panel). (B) Pooled data showing the frequencies of melanoma CD8+ TILs (n = 22) expressing PD-1 and/or TIGIT. P values were obtained by repeated-measures ANOVA, followed by Tukey’s multiple comparisons test. (C) Pooled data showing the frequencies of CD8+ TILs producing IFN-γ, TNF, or IL-2 according to PD-1 and TIGIT expression or PD-1 and TIM-3 coexpression (n = 9) after a 6-hour IVS with PMA and ionomycin. P values were obtained by repeated-measures ANOVA, followed by Tukey’s (left panel) and Friedman’s tests, followed by Dunn’s multiple comparisons test (center and right panels). (D) Pooled data showing the percentages of CFSElo and CD107a+ CD8+ TILs from melanoma patients (n = 6) after a 5-day IVS with IFN-γ–treated autologous CD3– cells and anti-CD3 Ab. P values were obtained by Wilcoxon’s test. *P < 0.05; **P < 0.01; ***P < 0.001. Horizontal bars depict the mean. Data are representative of 3 independent experiments.
Figure 6
Figure 6. The TIGIT ligands CD155 and CD112 are upregulated on metastatic melanoma cells and APCs in the TME.
(A) Frequencies of APCs and tumor cells within the CD3– cells isolated from metastatic melanoma single-cell suspensions (n = 11). (B) Representative flow cytometric analysis of CD155 and CD112 expression on CD11c–CD14–SSChiCSPG4+ melanoma cells, CD11c+CD14+ monocytes, and CD11c+CD14– DCs from CD3– cells from PBMCs from 1 healthy donor and 1 melanoma patient and from 1 metastatic melanoma single-cell suspension. (C) Pooled data showing CD155 (n = 16), CD112 (n = 10), PD-L1 (n = 20), HVEM (n = 17), and galectin-9 (n = 5) expression by CSPG4+ cells from melanoma single-cell suspensions. P values were obtained by Friedman’s, test followed by Dunn’s multiple comparisons test. (D) Pooled data showing CD155 and CD112 expression (percentage and MFI) on monocytes and DCs from PBMCs from healthy donors (n = 12) and melanoma patients (n = 8) and from metastatic melanoma single-cell suspensions (n = 15). P values were obtained by a 1-way ANOVA, followed by Tukey’s multiple comparisons test (bottom row, second panel from the left) and by a Kruskal-Wallis test, followed by Dunn’s multiple comparisons test (all other panels). Data are representative of 3 independent experiments. Horizontal bars depict the mean. *P < 0.05; **P < 0.01; ***P < 0.001.
Figure 5
Figure 5. PD-1 blockade increases TIGIT expression by NY-ESO-1–specific CD8+ T cells.
PBMCs from melanoma patients were incubated in vitro for 6 days with NY-ESO-1 157-165 peptide and blocking mAbs against TIGIT or PD-1 or isotype control mAbs (IgG). (A and B) Pooled data for TIGIT (n = 7) and PD-1 (n = 10) expression as shown by MFI (A) and fold-change of expression (B) on A2/NY-ESO-1 157-165 tet+ CD8+ T cells and tet– CD8+ T cells after PD-1 and TIGIT blockade, respectively. P = 0.016 by a 2-tailed, paired t test. Data are representative of 2 independent experiments performed in duplicate.
Figure 4
Figure 4. TIGIT blockade added to PD-1 blockade increases the frequencies of cytokine-producing NY-ESO-1–specific CD8+ T cells.
PBMCs from melanoma patients were incubated in vitro for 6 days with NY-ESO-1 157-165 peptide and blocking mAbs against TIGIT and/or PD-1 or isotype control mAbs (IgG) before evaluating intracellular cytokine production of A2/NY-ESO-1 157-165 tet+ CD8+ T cells in response to the cognate peptide. (A and B) Representative flow cytometric analysis of cells from 1 melanoma patient (A) and pooled data (n = 9) (B) showing the variation in numbers of IFN-γ– and TNF-producing NY-ESO-1 157-165 tet+ CD8+ T cells per 106 CD8+ T cells. P values were obtained by 2-tailed, paired t tests. (C) Fold-change of frequencies of IFN-γ– and TNF-producing A2/NY-ESO-1 157-165 tet+ CD8+ T cells after a 6-day IVS with cognate antigen and the indicated mAb (n = 9). P values were obtained by Friedman’s test, followed by Dunn’s multiple comparisons test (top panel) and by repeated-measures ANOVA (bottom panel), followed by Tukey’s multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001. Data shown are representative of 2 independent experiments performed in duplicate.
Figure 3
Figure 3. TIGIT blockade alone or in combination with PD-1 blockade increases the frequency of proliferating and total NY-ESO-1–specific CD8+ T cells.
CFSE-labeled PBMCs from melanoma patients were incubated in vitro for 6 days with NY-ESO-1 157-165 or HIV-pol 476-484 peptide and blocking mAbs against TIGIT (aTIGIT) and/or PD-1 (aPD-1) or isotype control mAbs (IgG). (A) Representative flow cytometric analysis for 1 melanoma patient showing the percentages of CFSElo A2/NY-ESO-1 157-165 tet+ CD8+ T cells among total CD8+ T cells. (B and C) Pooled data showing the variation in the numbers of CFSElo (B) and total (C) A2/NY-ESO-1 157-165 tet+ cells per 106 CD8+ T cells (n = 9). P values were obtained by 2-tailed, paired t tests. (D and E) Fold-change of the frequencies of CFSElo (D) and total (E) A2/NY-ESO-1 157-165 tet+ cells in the presence of the indicated blocking mAb (n = 9). P values were obtained by repeated-measures ANOVA, followed by Tukey’s multiple comparisons test. *P < 0.05. Data shown are representative of 2 independent experiments performed in duplicate.
Figure 2
Figure 2. TIGIT+PD-1+ NY-ESO-1–specific CD8+ T cells exhibit an effector memory and activated T cell phenotype.
(A and B) Representative dot plots for 1 melanoma patient (A) and summary data for 8 melanoma patients (B) indicating the frequencies (%) of CD38, HLA-DR, CD45RA, and CCR7 expressed on A2/NY-ESO-1 157-165 tet+ CD8+ T cells and tet– CD8+ T cells expressing TIGIT and/or PD-1. P values were obtained by Friedman’s test, followed by Dunn’s multiple comparisons test (top left panel) and by repeated-measures ANOVA, followed by Tukey’s multiple comparisons test (all other panels). (C) Ex vivo expression of TIGIT (MFI) on A2/NY-ESO-1–specific CD8+ T cells according to PD-1 and TIM-3 coexpression. P values were obtained by repeated-measures ANOVA, followed by Tukey’s multiple comparisons test. Horizontal bars depict the mean percentage or MFI. *P < 0.05; **P < 0.01; ***P < 0.001.
Figure 1
Figure 1. TIGIT is upregulated and coexpressed with PD-1 on NY-ESO-1–specific CD8+ T cells.
(A) Representative dot plots for 1 melanoma patient showing ex vivo TIGIT expression on A2/NY-ESO-1 157-165, A2/Flu-M 58-66, and A2/CMV 495-503 tet+ CD8+ T cells. CD8+ T cells stained with A2/HIV pol 476-484 tetramers or PE-labeled IgG control mAbs were used to establish the threshold for identifying tet+ and TIGIT+ cells, respectively. (B) Pooled data showing the percentage and MFI of TIGIT expression on NY-ESO-1–, Flu-, and CMV-specific CD8+ T cells as well as on total effector (CD45RA+CCR7–) and effector memory (CD45RO+CCR7–) CD8+ T cells from melanoma patients (n = 8). P values were obtained by repeated-measures ANOVA, followed by Tukey’s multiple comparisons test. (C) Dot plots for 1 representative melanoma patient showing ex vivo TIGIT and PD-1 expression on A2/NY-ESO-1 157-165, A2/Flu-M 58-66, and A2/CMV 495-503 tet+ CD8+ T cells as well as on total tet– CD8+ T cells. (D) Pooled data showing the distribution of NY-ESO-1–, Flu-, and CMV-specific CD8+ T cells, as well as of total effector and effector memory CD8+ T cells according to TIGIT and PD-1 expression in cells from melanoma patients (n = 8). P values were obtained by Friedman’s test, followed by Dunn’s multiple comparisons test. Horizontal bars depict the mean percentage or MFI. *P < 0.05; **P < 0.01; ***P < 0.001. Data shown are representative of 3 independent experiments.
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References

    1. Boon T, Coulie PG, Van den Eynde BJ, van der Bruggen P. Human T cell responses against melanoma. Annu Rev Immunol. 2006;24:175–208. doi: 10.1146/annurev.immunol.24.021605.090733. - DOI - PubMed
    1. Wherry EJ, Barber DL, Kaech SM, Blattman JN, Ahmed R. Antigen-independent memory CD8 T cells do not develop during chronic viral infection. Proc Natl Acad Sci U S A. 2004;101(45):16004–16009. doi: 10.1073/pnas.0407192101. - DOI - PMC - PubMed
    1. Wherry EJ. T cell exhaustion. Nat Immunol. 2011;12(6):492–499. - PubMed
    1. Ahmadzadeh M, et al. Tumor antigen-specific CD8 T cells infiltrating the tumor express high levels of PD-1 and are functionally impaired. Blood. 2009;114(8):1537–1544. doi: 10.1182/blood-2008-12-195792. - DOI - PMC - PubMed
    1. Fourcade J, et al. PD-1 is a regulator of NY-ESO-1-specific CD8+ T cell expansion in melanoma patients. J Immunol. 2009;182(9):5240–5249. doi: 10.4049/jimmunol.0803245. - DOI - PMC - PubMed

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