Development of mouse models with restricted HLA-B∗57:01 presentation for the study of flucloxacillin-driven T-cell activation and tolerance in liver injury

Ananthula, Suryatheja; Krishnavenisivakumar, Kirthiram; Cardone, Marco; Su, Shan; Roderiquez, Gregory; Abuzeineh, Hanan; Kleiner, David E.; Norcross, Michael A.; Puig, Montserrat

doi:10.1016/j.jaci.2023.03.029

Cited by 8 publications

(8 citation statements)

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“…With regard to adverse reactions to antibiotics, a flucloxacillin DILI mouse model was assessed. A key finding of this model was that hepatotoxic effects produced by CD8 + T cells were restricted by the tolerogenic environment in the liver and the absence of T reg cells may promote cytotoxic T cell responses through lack of IL-2 depletion ( 68 ). A humanized mouse model was used to evaluate IL-2-mediated toxicities in different organs.…”

Section: Discussionmentioning

confidence: 99%

Immuno-inflammatory in vitro hepatotoxicity models to assess side effects of biologicals exemplified by aldesleukin

Roser,

Luckhardt,

Ziegler

et al. 2023

Front. Immunol.

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IntroductionHepatotoxicity induced by immunotherapeutics is an appearing cause for immune-mediated drug-induced liver injury. Such immuno-toxic mechanisms are difficult to assess using current preclinical models and the incidence is too low to detect in clinical trials. As hepatotoxicity is a frequent reason for post-authorisation drug withdrawal, there is an urgent need for immuno-inflammatory in vitro models to assess the hepatotoxic potential of immuno-modulatory drug candidates. We developed several immuno-inflammatory hepatotoxicity test systems based on recombinant human interleukin-2 (aldesleukin).MethodsCo-culture models of primary human CD8+ T cells or NK cells with the hepatocyte cell line HepaRG were established and validated with primary human hepatocytes (PHHs). Subsequently, the HepaRG model was refined by increasing complexity by inclusion of monocyte-derived macrophages (MdMs). The main readouts were cytotoxicity, inflammatory mediator release, surface marker expression and specific hepatocyte functions.ResultsWe identified CD8+ T cells as possible mediators of aldesleukin-mediated hepatotoxicity, with MdMs being implicated in increased aldesleukin-induced inflammatory effects. In co-cultures of CD8+ T cells with MdMs and HepaRG cells, cytotoxicity was induced at intermediate/high aldesleukin concentrations and perforin was upregulated. A pro-inflammatory milieu was created measured by interleukin-6 (IL-6), c-reactive protein (CRP), interferon gamma (IFN-γ), and monocyte chemoattractant protein-1 (MCP-1) increase. NK cells responded to aldesleukin, however, only minor aldesleukin-induced cytotoxic effects were measured in co-cultures. Results obtained with HepaRG cells and with PHHs were comparable, especially regarding cytotoxicity, but high inter-donor variations limited meaningfulness of the PHH model.DiscussionThe in vitro test systems developed contribute to the understanding of potential key mechanisms in aldesleukin-mediated hepatotoxicity. In addition, they may aid assessment of immune-mediated hepatotoxicity during the development of novel immunotherapeutics.

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Section: Discussionmentioning

confidence: 99%

Immuno-inflammatory in vitro hepatotoxicity models to assess side effects of biologicals exemplified by aldesleukin

Roser,

Luckhardt,

Ziegler

et al. 2023

Front. Immunol.

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“…HLA expression on antigen presenting cells was required as well as CD28 signaling through engagement of co-stimulatory CD80/86 molecules on antigen presenting cells [5 ▪ ]. Similarly, CD8+ T cells from drug naïve HLA∗B:57:01 Tg mice responded to FLX in vitro after prolonged repeated activation of cells because of a lower frequency of drug reactive T cells [6 ▪ ] compared with the reported polyclonal ABC-induced response [7]. These results paralleled those shown in human peripheral blood mononuclear cells (PBMCs) expressing HLA-B∗57:01 [8–11], proving the value of expressing the HLA risk allele in Tg mice to study immune activation by drugs.…”

Section: Proof-of-conceptmentioning

confidence: 99%

“…In in vivo studies with different immunocompetent HLA Tg murine strains, administration of drugs associated with DHRs has proven to be insufficient to trigger clinical disease, although T cell activation was demonstrated [ 4 , 5 ▪ , 6 ▪ , 12 , 13 ]. The lack of clinical symptoms in drug-treated mice reflects drug tolerance as experienced by most human subjects treated with the same medicines, indicating that in mice, like in humans, environmental and host factors other than the HLA risk allele are involved in the development of drug pathology.…”

Section: Initial In Vivo Mouse Studies With Drugs ...mentioning

confidence: 99%

“…FLX is thought to be presented through both covalent and noncovalent mechanisms [ 9 – 11 , 25 ]. To elucidate the factors leading to FLX-DILI associated with HLA-B∗57:01, Ananthula and collaborators [ 6 ▪ ] treated HLA-B∗57:01 Tg mice with FLX by skin sensitization with retinoic acid followed by oral gavage to enhance generation and mobilization of drug-reactive cells to the gut-liver axis and induce liver injury. In immune-competent mice, treatment with FLX was not sufficient to activate T cells responses to the drug.…”

Section: Efforts To Break Immune-tolerance and Characterization Of Im...mentioning

confidence: 99%

“…Therefore, to gain knowledge on mechanisms leading to FLX-DILI when drug is presented by HLA-B:57:01, our group generated a strain that exclusively expresses the human allele [HLA Tg/ H2-K b D b KO mice (Tg/KO)] [ 27 ▪ ]. These animals had decreased numbers of CD8+ T cells compared to HLA Tg animals due to the H2-K b D b deletion [ 28 ] but increased expression of the HLA transgene per cell [ 6 ▪ ], thereby resulting in enrichment of drug-reactive T cells in the CD8+ T cell repertoire as seen in other models for nondrug antigens [ 29 ]. Tg/ KO mice treated with FLX and aCD4 Ab had increased levels of drug-induced PD1+CD8+ T cells in both lymphatic organs and liver (Fig.…”

Section: Efforts To Break Immune-tolerance and Characterization Of Im...mentioning

confidence: 99%

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Transgenic murine models for the study of drug hypersensitivity reactions linked to HLA-I molecules

Puig

Norcross

2023

Current Opinion in Allergy &Amp; Clinical Immunology

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Purpose of review Immune-mediated drug hypersensitivity reactions (DHRs) can be life-threatening and an impediment to drug development. Mechanism of disease studies are difficult to perform in humans. Here we review human leukocyte antigens class I (HLA-I) transgenic murine models and highlight how these systems have helped to elucidate drug-specific and host immune factors that initiate, propagate and control severe drug toxicities to skin and liver. Recent findings HLA transgenic mice have been developed and used to study immune-mediated drug reactions in vitro and in vivo. CD8+ T cells from HLA-B∗57:01-expressing mice respond strongly to abacavir (ABC) in vitro but have self-limited responses to drug exposure in vivo. Immune tolerance can be overcome by depleting regulatory T cells (Treg) allowing antigen-presenting dendritic cells to express CD80/86 costimulatory molecules and signal through CD28 on the CD8+ T cell. Depletion of Treg also removes competition for interleukin 2 (IL-2) to allow T cell expansion and differentiation. Fine tuning of responses depends on inhibitory checkpoint molecules such as PD-1. Improved mouse models express only HLA in the absence of PD-1. These models show enhanced liver injury to flucloxacillin (FLX) which depends on drug priming, CD4+ T cell depletion, and lack of PD-1 expression. Drug-specific HLA-restricted cytotoxic CD8+ T cells can infiltrate the liver but are suppressed by Kupffer and liver sinusoidal endothelial cells. Summary HLA-I transgenic mouse models are now available to study ABC, FLX and carbamazepine-induced adverse reactions. In vivo studies range from characterizing drug-antigen presentation, T cell activation, immune-regulatory molecules and cell-cell interaction pathways that are specifically involved in causing or controlling unwanted DHRs.

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Clues of HLAs, metabolic SNPs, and epigenetic factors in T cell-mediated drug hypersensitivity reactions

Molatefi,

Talebi,

Samei

et al. 2024

Heliyon

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Development of mouse models with restricted HLA-B∗57:01 presentation for the study of flucloxacillin-driven T-cell activation and tolerance in liver injury

Cited by 8 publications

References 48 publications

Immuno-inflammatory in vitro hepatotoxicity models to assess side effects of biologicals exemplified by aldesleukin

Immuno-inflammatory in vitro hepatotoxicity models to assess side effects of biologicals exemplified by aldesleukin

Transgenic murine models for the study of drug hypersensitivity reactions linked to HLA-I molecules

Clues of HLAs, metabolic SNPs, and epigenetic factors in T cell-mediated drug hypersensitivity reactions

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