VISTA is an inhibitory immune checkpoint that is increased after ipilimumab therapy in patients with prostate cancer

Journal name:
Nature Medicine
Volume:
23,
Pages:
551–555
Year published:
DOI:
doi:10.1038/nm.4308
Received
Accepted
Published online

To date, anti-CTLA-4 (ipilimumab) or anti-PD-1 (nivolumab) monotherapy has not been demonstrated to be of substantial clinical benefit in patients with prostate cancer. To identify additional immune-inhibitory pathways in the prostate-tumor microenvironment, we evaluated untreated and ipilimumab-treated tumors from patients in a presurgical clinical trial. Levels of the PD-L1 and VISTA inhibitory molecules increased on independent subsets of macrophages in treated tumors. Our data suggest that VISTA represents another compensatory inhibitory pathway in prostate tumors after ipilimumab therapy.

At a glance

Figures

  1. Treatment with ipilimumab increases immune cell infiltration, as well as expression of PD-L1 and VISTA in prostate tumors.
    Figure 1: Treatment with ipilimumab increases immune cell infiltration, as well as expression of PD-L1 and VISTA in prostate tumors.

    (a) Frequency of CD4+, CD8+, and ICOS+ T cells in untreated (n = 11) and treated (n = 6) tumors. (b) IHC analyses of CD4+, CD8+, ICOS+, CD45RO+, and GrB+ T cells, as well as CD68+ macrophages. (c) IHC of PD-L1, PD-1, and VISTA in tumor cells and tumor-infiltrating immune cells. In b and c, tumors are from three different cohorts of stage-matched patients: untreated (n = 18), treated with ADT (n = 10), and treated with ipilimumab (Ipi) + ADT (n = 16). An asterisk represents patients who received high-dose steroids with surgery delay. (d) Frequency of PD-L1 expression on CD4+ T cells, CD8+ T cells, CD68+ macrophages, and tumor cells. (e) Frequency of VISTA expression on CD4+ T cells, CD8+ T cells, and CD68+ macrophages. In d and e, matched pre-treatment (n = 10) and post-treatment (n = 10) tumors are shown. (f) IHC staining of CD4+ and CD8+ T cells and CD68+ macrophages in stage-matched untreated (n = 18) and ipilimumab- + ADT-treated (n = 15) prostate tumors as compared to stage-matched untreated (n = 18) and ipilimumab-treated (n = 20) metastatic melanomas. Arrows indicate a significant difference in CD8+ T cells and CD68+ macrophages between untreated prostate tumors and untreated melanomas. (g) Frequency of PD-L1 expression on CD4+ T cells, CD8+ T cells, and CD68+ macrophages. (h) Frequency of VISTA expression on CD4+ T cells, CD8+ T cells, and CD68+ macrophages. In g and h, matched pre-treatment (n = 10) and post-treatment (n = 10) prostate tumors are compared to matched pre-treatment (n = 10) and post-treatment (n = 10) melanomas. Pt, patient. In dh, the bars represent the mean of each group. P values were calculated using Welch's t-test.

  2. PD-L1+ and VISTA+ macrophages (CD68+) manifest an M2 phenotype and suppress T cell function.
    Figure 2: PD-L1+ and VISTA+ macrophages (CD68+) manifest an M2 phenotype and suppress T cell function.

    (a) Representative photographs from immunofluorescence (IF) multiplex staining in a total of nine post-treatment prostate tumors. Yellow, CD68; white, VISTA; purple, PD-L1; tumor nuclei are stained with DAPI (blue). Scale bars, 20 μm. (b) Frequency of PD-L1 and VISTA expression on CD68+ macrophages from stage-matched untreated (UnTx; n = 9) and post-treatment (Post; n = 9) prostate tumors. The bars represent the mean for each group. (c) Fold induction of M1-like genes in post-treatment prostate tumors (n = 6) and post-treatment melanomas (n = 20) as compared to untreated samples from the same types of tumors. (d) Quantitative IHC analysis of ARG1+ cells in untreated (n = 10) and post-treatment (n = 15) prostate tumors (left), as well as the ratio of ARG1 to iNOS in untreated and post-treatment tumors (right). Results represent the means ± s.d. for all samples in each group. (e) Representative photographs from multiplex IF staining of tumor nuclei (blue), CD68 (yellow), and CD163 (green) with PD-L1 (white) or VISTA (white) in post-treatment prostate tumors (left), as well as quantitative analysis of CD163 expression by CD68+PD-L1+ or CD68+VISTA+ cells from pre-treatment (n = 5) and matched post-treatment (n = 5) prostate tumors (right). Scale bars, 20 μm. (f) IFN-γ (left) and TNF-α (right) production by peripheral T cells from patients (n = 7) after CD3 stimulation (α-CD3) in the presence of plate-bound control immunoglobulin (Ctrl Ig), PD-L1 Ig, VISTA Ig, or a combination of PD-L1 Ig and VISTA Ig. *P < 0.05, **P < 0.001. (g) IFN-γ production by peripheral T cells from patients after CD3 stimulation (α-CD3) without coculture of monocytes or in the presence of either untreated monocytes (CD14+ + Ctrl IgG) or monocytes pretreated with anti-VISTA antibody (CD14+ + α-VISTA). Results represent the means ± s.d. for all samples in each group. Experiments were performed in triplicate. Pt, patient. P values were calculated using Welch's t-test.

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Author information

Affiliations

  1. Department of Genitourinary Medical Oncology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Jianjun Gao,
    • Lewis Z Shi,
    • Sumit K Subudhi,
    • Jianfeng Chen,
    • Eleni Efstathiou,
    • Christopher J Logothetis &
    • Padmanee Sharma
  2. Department of Urology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • John F Ward &
    • Curtis A Pettaway
  3. The Immunotherapy Platform, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Luis M Vence,
    • Hao Zhao,
    • Hong Chen,
    • James P Allison,
    • Jingjing Sun,
    • Jorge Blando &
    • Padmanee Sharma
  4. Department of Pathology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Patricia Troncoso
  5. Department of Immunology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • James P Allison &
    • Padmanee Sharma
  6. Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Ignacio I Wistuba
  7. Janssen Oncology Therapeutic Area, Janssen Research and Development, LLC, Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania, USA.

    • Manuel A Sepulveda
  8. Department of Surgical Oncology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Jennifer Wargo

Contributions

P.S. designed the study; L.Z.S., J.B., L.M.V., J.C., I.I.W., M.A.S., J.S., and H.C. performed the experiments; J.G., J.F.W., C.A.P., L.Z.S., J.C., J.B., L.M.V., I.I.W., J.S., H.Z., J.W., E.E., and P.T. analyzed the data; J.G., P.S., J.P.A., S.K.S., J.W., and C.J.L. drafted the manuscript; and all authors reviewed the final draft of the manuscript.

Competing financial interests

P.S. and J.P.A. are founders and advisors for Jounce Therapeutics. P.S. and J.P.A. are members of the Parker Institute for Cancer Immunotherapy. P.S. also serves as a consultant for BMS, AstraZeneca, Amgen, and GlaxoSmithKline. J.P.A. is an inventor of intellectual property owned by the University of California, Berkeley, and licensed to BMS and has received royalties from BMS. J.P.A. is also inventor of intellectual property owned by Memorial Sloan Kettering Cancer Center and licensed to Merck. J.G. serves as a consultant for Genentech. I.I.W. serves as a consultant for BMS. E.E. serves as a consultant for Janssen, Bayer, Medivation, Astellas, and Sanofi Takeda.

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