A transgenic mouse model for HLA-B*57:01–linked abacavir drug tolerance and reactivity
Marco Cardone, … , David H. Margulies, Michael A. Norcross
Marco Cardone, … , David H. Margulies, Michael A. Norcross
Published May 21, 2018
Citation Information: J Clin Invest. 2018;128(7):2819-2832. https://doi.org/10.1172/JCI99321.
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Research Article Immunology

A transgenic mouse model for HLA-B*57:01–linked abacavir drug tolerance and reactivity

Abstract

Adverse drug reactions (ADRs) are a major obstacle to drug development, and some of these, including hypersensitivity reactions to the HIV reverse transcriptase inhibitor abacavir (ABC), are associated with HLA alleles, particularly HLA-B*57:01. However, not all HLA-B*57:01+ patients develop ADRs, suggesting that in addition to the HLA genetic risk, other factors may influence the outcome of the response to the drug. To study HLA-linked ADRs in vivo, we generated HLA-B*57:01–Tg mice and show that, although ABC activated Tg mouse CD8+ T cells in vitro in a HLA-B*57:01–dependent manner, the drug was tolerated in vivo. In immunocompetent Tg animals, ABC induced CD8+ T cells with an anergy-like phenotype that did not lead to ADRs. In contrast, in vivo depletion of CD4+ T cells prior to ABC administration enhanced DC maturation to induce systemic ABC-reactive CD8+ T cells with an effector-like and skin-homing phenotype along with CD8+ infiltration and inflammation in drug-sensitized skin. B7 costimulatory molecule blockade prevented CD8+ T cell activation. These Tg mice provide a model for ABC tolerance and for the generation of HLA-B*57:01–restricted, ABC-reactive CD8+ T cells dependent on both HLA genetic risk and immunoregulatory host factors.

Authors

Marco Cardone, Karla Garcia, Mulualem E. Tilahun, Lisa F. Boyd, Sintayehu Gebreyohannes, Masahide Yano, Gregory Roderiquez, Adovi D. Akue, Leslie Juengst, Elliot Mattson, Suryatheja Ananthula, Kannan Natarajan, Montserrat Puig, David H. Margulies, Michael A. Norcross

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Figure 2

CD4+ T cells prevent ABC drug reactivity in HLA-B*57:01–Tg mice.

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CD4+ T cells prevent ABC drug reactivity in HLA-B*57:01–Tg mice. HLA-B*5...
HLA-B*57:01–Tg or WT mice were treated systemically (i.p. injection) and topically (ear painting) with vehicle (Veh) or ABC, in the absence or presence of a CD4-depleting mAb. (A) Photos of ears (left) and CD8α staining of ear sections (IHC, right) from Tg mice treated for 3 weeks. Data are representative of 2 independent experiments. (B) Percentage of PD-1+ cells within CD8+ T lymphocytes in the LNs of treated Tg mice, as measured by flow cytometry. (C) Percentage of PD-1+, Ki-67+, and BrdU+ cells within CD8+ T lymphocytes in the LNs of treated Tg mice. Flow cytometric data are from 1 of 2 experiments. (D) Percentage of CD44- and CD62L-expressing cells within CD8+PD-1+ T lymphocytes in the LNs of ABC-exposed Tg mice, as measured by flow cytometry. n = 3–6 mice per time point. Statistics refer to the comparison of CD44hiCD62Lhi versus CD44hiCD62Llo cells. (E) IFN-γ in supernatants from day 5 cultures of CD8+ T cells from the LNs of ABC-naive or -treated Tg animals, as measured by ELISA. (F) Photos of ears (left) and CD8α staining of ear sections (IHC, right) from CD4-depleted Tg mice treated for 3 weeks. Data are representative of 2 independent experiments. (G) Ear thickness at week 3 of treatment. (H) Percentage of PD-1+ cells within CD8+ T lymphocytes in the LNs of Tg mice, as measured by flow cytometry at day 10 of treatment. Animals in the ABC control group were also included in the ABC (day 10) group in B. Scale bars: 100 μm. Data represent the mean ± SEM. Dots indicate values for individual mice from each group: n = 3–11 (B); n = 3–10 (E); n = 4–12 (G); n = 4–7 (H). *P < 0.05, **P < 0.005, ***P < 0.0005, and ****P < 0.0001, by unpaired, 2-tailed Student’s t test (B and E), 2-way ANOVA (D), or 1-way ANOVA (G and H) with Tukey’s multiple comparisons correction. None, no drug.