PMID- 33633747 OWN - NLM STAT- MEDLINE DCOM- 20210623 LR - 20210623 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 11 DP - 2020 TI - Alterations in the HLA-B*57:01 Immunopeptidome by Flucloxacillin and Immunogenicity of Drug-Haptenated Peptides. PG - 629399 LID - 10.3389/fimmu.2020.629399 [doi] LID - 629399 AB - Neoantigen formation due to the interaction of drug molecules with human leukocyte antigen (HLA)-peptide complexes can lead to severe hypersensitivity reactions. Flucloxacillin (FLX), a beta-lactam antibiotic for narrow-spectrum gram-positive bacterial infections, has been associated with severe immune-mediated drug-induced liver injury caused by an influx of T-lymphocytes targeting liver cells potentially recognizing drug-haptenated peptides in the context of HLA-B*57:01. To identify immunopeptidome changes that could lead to drug-driven immunogenicity, we used mass spectrometry to characterize the proteome and immunopeptidome of B-lymphoblastoid cells solely expressing HLA-B*57:01 as MHC-I molecules. Selected drug-conjugated peptides identified in these cells were synthesized and tested for their immunogenicity in HLA-B*57:01-transgenic mice. T cell responses were evaluated in vitro by immune assays. The immunopeptidome of FLX-treated cells was more diverse than that of untreated cells, enriched with peptides containing carboxy-terminal tryptophan and FLX-haptenated lysine residues on peptides. Selected FLX-modified peptides with drug on P4 and P6 induced drug-specific CD8(+) T cells in vivo. FLX was also found directly linked to the HLA K146 that could interfere with KIR-3DL or peptide interactions. These studies identify a novel effect of antibiotics to alter anchor residue frequencies in HLA-presented peptides which may impact drug-induced inflammation. Covalent FLX-modified lysines on peptides mapped drug-specific immunogenicity primarily at P4 and P6 suggesting these peptide sites as drivers of off-target adverse reactions mediated by FLX. FLX modifications on HLA-B*57:01-exposed lysines may also impact interactions with KIR or TCR and subsequent NK and T cell function. CI - Copyright (c) 2021 Puig, Ananthula, Venna, Kumar Polumuri, Mattson, Walker, Cardone, Takahashi, Su, Boyd, Natarajan, Abdoulaeva, Wu, Roderiquez, Hildebrand, Beaucage, Li, Margulies and Norcross. FAU - Puig, Montserrat AU - Puig M AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Ananthula, Suryatheja AU - Ananthula S AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Venna, Ramesh AU - Venna R AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Kumar Polumuri, Swamy AU - Kumar Polumuri S AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Mattson, Elliot AU - Mattson E AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Walker, Lacey M AU - Walker LM AD - Division of Applied Regulatory Science, Office of Translational Science, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Cardone, Marco AU - Cardone M AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Takahashi, Mayumi AU - Takahashi M AD - Laboratory of Biological Chemistry, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Su, Shan AU - Su S AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Boyd, Lisa F AU - Boyd LF AD - Molecular Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States. FAU - Natarajan, Kannan AU - Natarajan K AD - Molecular Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States. FAU - Abdoulaeva, Galina AU - Abdoulaeva G AD - Facility for Biotechnology Resources, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Wu, Wells W AU - Wu WW AD - Facility for Biotechnology Resources, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Roderiquez, Gregory AU - Roderiquez G AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Hildebrand, William H AU - Hildebrand WH AD - Department of Microbiology and Immunology, School of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, United States. FAU - Beaucage, Serge L AU - Beaucage SL AD - Laboratory of Biological Chemistry, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Li, Zhihua AU - Li Z AD - Division of Applied Regulatory Science, Office of Translational Science, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. FAU - Margulies, David H AU - Margulies DH AD - Molecular Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States. FAU - Norcross, Michael A AU - Norcross MA AD - Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States. LA - eng PT - Journal Article PT - Research Support, N.I.H., Intramural PT - Research Support, U.S. Gov't, Non-P.H.S. PT - Research Support, U.S. Gov't, P.H.S. DEP - 20210209 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - 0 (HLA-B Antigens) RN - 0 (HLA-B*57:01 antigen) RN - 0 (Haptens) RN - 0 (Peptides) RN - 43B2M34G2V (Floxacillin) SB - IM MH - Animals MH - Cell Line MH - Floxacillin/*immunology MH - HLA-B Antigens/genetics/*immunology MH - Haptens/*immunology MH - Humans MH - Mice MH - Mice, Transgenic MH - Peptides/genetics/*immunology PMC - PMC7900192 OTO - NOTNLM OT - HLA-B*57:01 OT - drug hypersensitivity OT - flucloxacillin OT - hapten OT - immunogenicity OT - transgenic mice COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2021/02/27 06:00 MHDA- 2021/06/24 06:00 PMCR- 2020/01/01 CRDT- 2021/02/26 06:03 PHST- 2020/11/14 00:00 [received] PHST- 2020/12/23 00:00 [accepted] PHST- 2021/02/26 06:03 [entrez] PHST- 2021/02/27 06:00 [pubmed] PHST- 2021/06/24 06:00 [medline] PHST- 2020/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2020.629399 [doi] PST - epublish SO - Front Immunol. 2021 Feb 9;11:629399. doi: 10.3389/fimmu.2020.629399. eCollection 2020.