Insights into regulatory T-cell and type-I interferon roles in determining abacavir-induced hypersensitivity or immune tolerance

Marco Cardone¹Hratch M Baghdassarian²Maryam Khalaj¹Kirthiram Krishnaveni Sivakumar¹SuJin Hwang¹Sintayehu Gebreyohannes¹KAZUYO TAKEDA¹Yura Jang¹Nathan E Lewis^2,3Michael A Norcross¹Montserrat Puig^1*

• ¹United States Food and Drug Administration, Silver Spring, Maryland, United States
• ²University of California, San Diego, La Jolla, California, United States
• ³Technical University of Denmark, Kongens Lyngby, Denmark

Introduction: Clinical use of several small molecule drugs may lead to severe T-cell-mediated idiosyncratic drug hypersensitivity reactions (iDHR) linked to HLA alleles, including abacavir (ABC) with HLA-B*57:01. Due to study limitations in humans, pathogenic networks in iDHR remain elusive. HLA transgenic murine models have been proposed to bridge knowledge gaps in tolerance and susceptibility to drugs. Methods: Mice expressing HLA-B*57:01 and Foxp3-DTR/EGFP were generated to selectively deplete regulatory T-cells (Treg) with diphtheria toxin. ABC was administered for 8 days alone or together with cell-and cytokine-depleting antibodies. Cellular and transcriptomic responses were analyzed by RNA, flow cytometry and fluorescence methods. Results: While CD8 + T-cell responses to ABC require HLA presentation, ABC also triggered mitochondrial stress in macrophages in vitro, independently of HLA. In vivo, Treg were the primary mechanism of drug tolerance controlling HLA presentation and costimulation by antigen presenting cells. Treg ablation uncovered immune adverse events linked to activation and proliferation of both drug-specific and bystander CD8 + T-cells through CD28-mediated pathways with support from CD4 + non-Treg. Type-I interferon (IFN-I) and cellular-stress pathways influenced the fate of lymph node cells responding to ABC, implicating innate immune cells such as macrophages and plasmacytoid dendritic cells in the development of T-cell responses against the drug. IFN-I and IL-2 were necessary for CD8 + T-cell differentiation and ABC-induced adverse reactions. Conclusions: This study unveils novel immune mechanisms driven by drug and host-related factors required for in vivo reactions and sheds light on potential biomarker and therapeutic targets for managing and preventing severe and life-threatening iDHR.