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 1

ABC activates Tg mouse CD8+ T cells in vitro in an HLA-B*57:01–dependent manner.

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ABC activates Tg mouse CD8+ T cells in vitro in an HLA-B*57:01–dependent...
Cultures of purified splenic CD8+ T cells and cultures of total LN cells from drug-naive HLA-B*57:01–Tg (B*57:01) or WT mice. (A) Bright-field microscopy images (original magnification, ×10) of purified CD8+ T cells on day 5 of culture. Data are from 1 of 3 representative experiments. (B) IL-2, IFN-γ, and granzyme B (GZB) in supernatants of purified CD8+ T cells at the indicated time of culture. Data represent the mean ± SEM of ELISA results. Dots indicate the averages of technical replicates in each condition within individual experiments (n = 3–7 experiments). (C and D) Percentage of PD-1+, CD25+, and IFN-γ+ cells within CD8+ (C) and CD4+ (D) T lymphocytes in Tg purified CD8+ T cells and total LN cells cultured for 5 days. Flow cytometric data are from 1 of 2 representative experiments. (E) IFN-γ release by ABC-reactive CD8+ T cells restimulated with 5 μg/ml ABC, in the absence or presence of the specified mAb, following 14 days of primary stimulation. IFN-γ enzyme-linked immunosorbent spot (ELISpot) data show 4 replicates per condition from 1 of 3 representative experiments. *P < 0.05 and **P < 0.005, by unpaired, 2-tailed Student’s t test. None, no drug.