Research ArticleAutoimmunity

Antibody blockade of IL-15 signaling has the potential to durably reverse vitiligo

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Science Translational Medicine  18 Jul 2018:
Vol. 10, Issue 450, eaam7710
DOI: 10.1126/scitranslmed.aam7710
  • Fig. 1 Melanocyte-specific CD8+ TRM are present in vitiligo patient skin and express the CD122 chain of the IL-15 receptor, whereas keratinocytes express the CD215 chain.

    (A) Sample image of a vitiligo skin blister biopsy. (B) Flow cytometry staining of blister fluid in a vitiligo patient pregated on live single CD45+CD8+ T cells. MART-1, gp100, and tyrosinase pentamers were pooled to identify melanocyte antigen reactive cells, and TRM phenotype markers were assessed using CD69 and CD103 antibodies. (C) Quantification of the frequency of melanocyte-specific cells with a TRM phenotype in peripheral blood mononuclear cells (PBMCs) and lesional and nonlesional skin. Melanocyte-specific TRM were significantly enriched in lesional skin. [Each symbol represents one patient; open squares were stained with MART-1 pentamer alone, and closed circles were stained with all three pentamers MART-1, gp100, and tyrosinase; one-way analysis of variance (ANOVA) with Tukey’s post tests significant as indicated.] (D) Flow cytometry staining of CD122 on pentamer+ and pentamer TRM and (E) quantification in PBMCs and lesional and nonlesional skin. Cells were pregated on live single CD45+CD8+CD69+CD103+pentamer+/− T cells [two-way ANOVA for %CD122+, P = 0.0094 significant for differences between pentamer+ and pentamer cells; post tests, not significant (ns); two-way ANOVA for median fluorescence intensity (MFI) CD122, ns; paired t test, P = 0.0154 for PBMCs]. (F) Flow cytometry staining of CD215 on keratinocytes from blister roofs and (G) quantification in lesional and nonlesional skin. Cells were pregated on live single CD45 cells. CD215 expression was more frequent in lesional than nonlesional skin (Student’s t tests significant as indicated, P = 0.0385). FMO, Flourescence Minus One control.

  • Fig. 2 Melanocyte-specific T cells exhibit a TRM phenotype and express CD122 in the vitiligo mouse model.

    (A) Sample flow plots examining CD69 and CD103 expression on premelanosome protein (gp100)–specific transgenic T cells (PMEL) from the epidermis of vitiligo mice at weeks 3, 5, 7, 8, and 27 after vitiligo induction. Cells were pregated on live single CD45+CD8+ and Thy1.1+ or green fluorescent protein–positive (GFP+) to identify transferred PMEL cells. (B) Quantification of the total number of PMEL in the epidermis (left axis, black line) and the frequency of CD69+CD103+ PMEL TRM in the epidermis (right axis, blue line) over time after vitiligo induction (n = 4 mice at week 3, 5 mice at week 5, 5 mice at week 7, 2 mice at week 8, 3 mice at week 17, 4 mice at week 27, and 2 mice at week 63 pooled from four separate experiments). (C) Representative flow cytometry staining of CD122 on PMEL and host CD8+ TRM and (D) quantification in mouse epidermis at week 8 (Student’s t test significant as indicated, P = 0.0009 and P = 0.0018; n = 5 mice pooled from two representative experiments).

  • Fig. 3 Treatment with anti-CD122 antibody reverses disease in mice with established vitiligo.

    (A) Timing of treatments in the efficacy/repigmentation model. i.v., intravenous; i.p., intraperitoneal. (B) Sample photos of vehicle control [phosphate-buffered saline (PBS) or isotype] and anti-CD122 antibody (Ab)–treated animals at treatment baseline and week 8. (C) Comparison of the final percent change in pigmentation in vehicle and anti-CD122 antibody–treated animals. (D) Quantification of PMEL numbers in treated animals within the indicated tissues. (E) Quantification of host CD8+ T cell numbers in treated animals within the indicated tissues (each dot represents one animal; pooled from two separate experiments, n = 11 vehicle mice and n = 11 anti-CD122 antibody–treated mice; t tests significant as indicated). LN, lymph node.

  • Fig. 4 Systemic anti-CD122 antibody treatment durably reverses disease in mice via inhibition of antigen-specific T cell IFNγ production.

    (A) Timing of treatments in the systemic durability study. (B) Sample photos of vehicle control (PBS or isotype) and anti-CD122 antibody–treated animals at treatment baseline and week 8. (C) Comparison of the final percent change in pigmentation in vehicle and CD122 antibody–treated animals (each dot represents one animal; pooled from three separate experiments, n = 12 control mice and n = 13 anti-CD122 antibody–treated mice; t test significant as indicated). (D) Analysis of the percent of tail with pigmentation over time (two-way ANOVA, P = 0.0015 for treatment, P < 0.0001 for time, and ns for interaction with Dunnett’s comparisons to baseline pigmentation with main time effect significant as indicated by asterisks; baseline versus week 6, P = 0.0143; baseline versus week 8, P = 0.0105; baseline versus week 10, P = 0.0001). (E) Timing of the treatments in the functionality study. (F) Sample flow plots of epidermal PMEL production of granzyme B and IFNγ. (G) Quantification of PMEL producing IFNγ in the indicated tissues (Student’s t tests significant as indicated; n = 14 vehicle mice and n = 14 anti-CD122 antibody–treated mice; pooled from three separate experiments).

  • Fig. 5 Local intradermal injection of anti-CD122 antibody treatment durably reverses disease in mice.

    (A) Timing of treatments in the local durability study. i.d., intradermal. (B) Sample photos of vehicle control (PBS or isotype) and CD122 antibody–treated animals at treatment baseline and week 12. (C) Comparison of the final percent change in pigmentation in control and CD122 antibody–treated animals (each dot represents one animal pooled from two separate experiments, n = 7 control mice and n = 9 CD122 antibody–treated mice; t test significant as indicated). (D) Analysis of the percent of tail with pigmentation over time (two-way ANOVA, ns for treatment and time, P = 0.0017 for interaction with Dunnett’s comparisons to baseline pigmentation with simple effects within treatment groups significant as indicated by asterisks; baseline versus week 10, P = 0.0197; baseline versus week 12, P = 0.0002).

Supplementary Materials

  • www.sciencetranslationalmedicine.org/cgi/content/full/10/450/eaam7710/DC1

    Fig. S1. Analysis of single versus pooled pentamer stains.

    Fig. S2. Analysis of melanocyte-specific cells in blister biopsies from healthy controls.

    Fig. S3. CD215 staining on human and mouse melanocyte-specific T cells.

    Fig. S4. Examining CD215 expression on dendritic cells in vitiligo patient blister fluid.

    Fig. S5. Immunohistochemistry of CD215 in vitiligo skin biopsies.

    Fig. S6. CD122 antibody blocks IL-15 activity, but not IL-2 activity, in mouse T cell bioassays.

    Fig. S7. Analysis of PMEL T cell populations in mice treated with intradermal CD122 antibody.

    Fig. S8. Proposed model for CD122 and IL-15/CD215 biology in vitiligo.

    Table S1. Characteristics of patients used in this study.

    Table S2. PMEL analysis in CD122 antibody systemic durability studies.

    Table S3. Primary data.

  • The PDF file includes:

    • Fig. S1. Analysis of single versus pooled pentamer stains.
    • Fig. S2. Analysis of melanocyte-specific cells in blister biopsies from healthy controls.
    • Fig. S3. CD215 staining on human and mouse melanocyte-specific T cells.
    • Fig. S4. Examining CD215 expression on dendritic cells in vitiligo patient blister fluid.
    • Fig. S5. Immunohistochemistry of CD215 in vitiligo skin biopsies.
    • Fig. S6. CD122 antibody blocks IL-15 activity, but not IL-2 activity, in mouse T cell bioassays.
    • Fig. S7. Analysis of PMEL T cell populations in mice treated with intradermal CD122 antibody.
    • Fig. S8. Proposed model for CD122 and IL-15/CD215 biology in vitiligo.
    • Table S1. Characteristics of patients used in this study.
    • Table S2. PMEL analysis in CD122 antibody systemic durability studies.
    • Legend for table S3

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    Other Supplementary Material for this manuscript includes the following:

    • Table S3. (Microsoft Excel format). Primary data.

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