Puig Montserrat, Ananthula Suryatheja, Venna Ramesh, Kumar Polumuri Swamy, Mattson Elliot, Walker Lacey M, Cardone Marco, Takahashi Mayumi, Su Shan, Boyd Lisa F, Natarajan Kannan, Abdoulaeva Galina, Wu Wells W, Roderiquez Gregory, Hildebrand William H, Beaucage Serge L, Li Zhihua, Margulies David H, Norcross Michael A
Laboratory of Immunology, Office of Biotechnology Products, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States.
Division of Applied Regulatory Science, Office of Translational Science, Center for Drugs Evaluation and Research, Food and Drug Administration, Silver Spring, MD, United States.
Front Immunol. 2021 Feb 9;11:629399. doi: 10.3389/fimmu.2020.629399. eCollection 2020.
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 β-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-B57:01. 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-B57:01 as MHC-I molecules. Selected drug-conjugated peptides identified in these cells were synthesized and tested for their immunogenicity in HLA-B57:01-transgenic mice. T cell responses were evaluated 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 . 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-B57:01-exposed lysines may also impact interactions with KIR or TCR and subsequent NK and T cell function.
药物分子与人类白细胞抗原（HLA）-肽复合物相互作用导致的新抗原形成可引发严重的超敏反应。氟氯西林（FLX）是一种用于窄谱革兰氏阳性菌感染的β-内酰胺类抗生素，与严重的免疫介导药物性肝损伤有关，这种损伤是由靶向肝细胞的T淋巴细胞流入引起的，这些T淋巴细胞可能在HLA-B57:01的背景下识别药物半抗原化肽段。为了确定可能导致药物驱动免疫原性的免疫肽组变化，我们使用质谱对仅表达HLA-B57:01作为MHC-I分子的B淋巴母细胞的蛋白质组和免疫肽组进行了表征。在这些细胞中鉴定出的选定药物缀合肽被合成，并在HLA-B57:01转基因小鼠中测试其免疫原性。通过免疫测定评估T细胞反应。氟氯西林处理细胞的免疫肽组比未处理细胞的免疫肽组更多样化，富含在肽段上含有羧基末端色氨酸和氟氯西林半抗原化赖氨酸残基的肽段。在P4和P6位置带有药物的选定氟氯西林修饰肽诱导了药物特异性CD8 T细胞。还发现氟氯西林直接与HLA K146相连，这可能会干扰KIR-3DL或肽段相互作用。这些研究确定了抗生素的一种新作用，即改变HLA呈递肽段中的锚定残基频率，这可能会影响药物诱导的炎症。肽段上共价氟氯西林修饰的赖氨酸主要在P4和P6位置确定了药物特异性免疫原性，表明这些肽段位点是氟氯西林介导的脱靶不良反应的驱动因素。HLA-B57:01暴露的赖氨酸上的氟氯西林修饰也可能影响与KIR或TCR的相互作用以及随后的NK和T细胞功能。
