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. 2008 Jun 10;105(23):8061-6.
doi: 10.1073/pnas.0710929105. Epub 2008 Jun 3.

Effective tumor treatment targeting a melanoma/melanocyte-associated antigen triggers severe ocular autoimmunity

Douglas C Palmer  1 , Chi-Chao Chan, Luca Gattinoni, Claudia Wrzesinski, Chrystal M Paulos, Christian S Hinrichs, Daniel J Powell Jr, Christopher A Klebanoff, Steven E Finkelstein, Robert N Fariss, Zhiya Yu, Robert B Nussenblatt, Steven A Rosenberg, Nicholas P Restifo
Affiliations

Effective tumor treatment targeting a melanoma/melanocyte-associated antigen triggers severe ocular autoimmunity

Douglas C Palmer et al. Proc Natl Acad Sci U S A. .

Abstract

Nonmutated tissue differentiation antigens expressed by tumors are attractive targets for cancer immunotherapy, but the consequences of a highly effective antitumor immune response on self-tissue have not been fully characterized. We found that the infusion of ex vivo expanded adoptively transferred melanoma/melanocyte-specific CD8+ T cells that mediated robust tumor killing also induced autoimmune destruction of melanocytes in the eye. This severe autoimmunity was associated with the up-regulation of MHC class I molecules in the eye and high levels of IFN-gamma derived from both adoptively transferred CD8+ T cells and host cells. Furthermore, ocular autoimmunity required the presence of the IFN-gamma receptor on target tissues. Data compiled from >200 eyes and tumors in 10 independently performed experiments revealed a highly significant correlation (P < 0.0001) between the efficacy of tumor immunotherapy and the severity of ocular autoimmunity. Administration of high doses of steroids locally mitigated ocular autoimmunity without impairing the antitumor effect. These findings have particular importance for immunotherapies directed against self-antigens and highlight the need for targeting unique tumor antigens not expressed in critical tissues.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Induction of ocular autoimmunity during effective tumor destruction. (A) Flow cytometric analysis of Vβ13+CD8+ MDA-reactive pmel-1 T cells in the inguinal lymph node or eye 5 days after exogenous IL-2 and recombinant poxviral immunization of pmel-1 (TCR+) or WT (TCR−) mice. (B) Eyes from A were H&E-stained and examined for changes in morphology of the cornea (i), iris (ii), photoreceptors (iii), or choroid (iv). Arrows highlight cellular infiltrates in pmel-1 mice receiving recombinant gp100 poxvirus and IL-2. Data are representative of two independently identically performed experiments. (C) Ocular autoimmunity at day 5 after IL-2 and vaccination were assessed by using a masked ocular autoimmunity score as described in Methods from two independently performed experiments, *, P > 0.0001 vs. pmel-1 without vaccination. (D) Treatment of established tumors in pmel-1 transgenics vaccinated with exogenous IL-2 and recombinant gp100 poxvirus (□), without vaccination (■), or vaccinated WT littermates (▴), representative of two independent experiments.
Fig. 2.
Fig. 2.
Ocular autoimmunity is associated with the presence of MDA-reactive CD8+ T cells in mice and humans. (A) Eyes from mice that received vaccination with or without the adoptive transfer of congenic pmel-1 Thy1.1+ T cells (P) were sectioned and stained with isotype (α IgG) or α Thy1.1 antibodies using immunohistochemistry. (B) Increase in ocular autoimmunity with escalating numbers of adoptively transferred pmel-1 at 0 (♦), 1 × 105 (●), 1 × 106 (▴), or 9 × 106 (■) T cells per mouse (n = 3 mice per group) with vaccination. (C) Anterior ocular inflammation in a metastatic melanoma patient after lymphodepletion then adoptive transfer of TIL and IL-2. (D) Flow cytometric analysis and enumeration of Mart-1/Melan-A tetramer positive CD8+ T cells from this patient's PBL after ACT.
Fig. 3.
Fig. 3.
Ocular and tumor immunity is CD8+ T cell-dependent. (A) Evaluation of ocular autoimmunity 15 days after ACT of CD8+ enriched pmel-1 T cells into Rag1−/− T and B cell-deficient mice and CD8 depletion (100 μg per mouse) at times indicated. (B) Evaluation of tumor growth after CD8 depletion at days: 1 (■), 5 (♦), 9 (▴), isotype (○), no antibody (●), or no pmel-1 (X) (n = 5 mice per group). Data are representative of two independently performed experiments.
Fig. 4.
Fig. 4.
Ocular and tumor immunity is associated with IFN-γ and the up-regulation of MHC class I. (A) In situ detection of pmel-1, IFN-γ, and MHC class I in the eye after ACT using confocal microscopy. (B) Detection of enhanced MHC class I expression in tumor 5 days after ACT of pmel-1 using confocal microscopy. Images are representative of multiple fields. (C) The induction of ocular autoimmunity was blindly evaluated 5 days after the adoptive transfer of WT, FasL, TNF-α, perforin or IFN-γ-deficient pmel-1 T cells in WT tumor-bearing mice. (D) Tumor growth was assessed after the ACT of pmel-1 WT T cells (●) or pmel-1 T cells deficient in IFN-γ (▴), perforin (▵), FasL (♦), TNF-α (■), or no cells (○). Data are representative of two independently performed experiments.
Fig. 5.
Fig. 5.
Ocular autoimmunity requires the presence of IFN-γ-receptor on host tissue. (A and B) Pmel-1 and host IFN-γ-dependent ocular and tumor immunity. (A) Assessment of ocular immunity after the ACT of pmel-1 WT or IFN-γ −/− T cells into WT or IFN-γ −/− recipients. (B) Tumor therapy after the ACT of pmel-1 WT into B6 (●) or IFN-γ− −/− (□) hosts, ACT of pmel-1 IFN-γ− −/− into B6 (◇) or IFN-γ −/− (▴) hosts, no cells B6 (X) or IFN-γ−/− (○) hosts. (C) Tumor therapy after the ACT of pmel-1 T cells and the administration IFN-γ neutralizing antibody (■), isotype (▴), PBS (○), or no cells (□). (D) Target tissue expression of IFN-γ receptor, but not TNF-α or Fas receptor, dictates immunity in the eye. Evaluation of ocular autoimmunity in B6 bone marrow reconstituted WT, IFN-γR, TNF-αR, or FasR-deficient chimeric recipients 5 days after ACT with pmel-1 (■) or no cells (□). Each group contains at least five mice per group, and data are representative of at least two independently performed experiments.
Fig. 6.
Fig. 6.
Improved tumor therapy correlates with the severity of ocular autoimmunity. (A) Mean ocular immunity score (day 5) vs. mean tumor slope for each group. Data are representative of 10 independent experiments, 46 treatment groups, 218 eyes, and >250 tumors. (B) Ablation augments ocular and tumor immunity. Pmel-1 ACT with (♦) or without (■) prior ablation or no pmel-1 with (▴) or without prior ablation (●). n = 56 mice, combination of two independent experiments.
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