Conjugation of human serum albumin and flucloxacillin provokes specific immune response in HLA-B*57:01 transgenic mice
Conjugation of human serum albumin and flucloxacillin provokes specific immune response in HLA-B*57:01 transgenic mice
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DOI：
10.1016/j.imlet.2022.08.002
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发表时间：
2022-08-22
影响因子：
4.4
通讯作者：
Ito, Kousei
中科院分区：
文献类型：
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作者：
Gao, Yuying;Song, Binbin;Ito, Kousei
氟氯西林（FLX）可引起肝脏不良反应，有报道称其与人白细胞抗原(HLA)-B*57:01有关。在之前的一项研究中，abacvir诱导的超敏反应被诱导到HLA-B*57:01转基因小鼠（B*57:01-Tg）中，该转基因小鼠最初是由我们的团队构建的（Susukida et al., 2021）。本研究采用B*57:01-Tg小鼠复制flx诱导的肝损伤。然而，单独用FLX治疗B*57:01-Tg小鼠并没有增加血清ALT水平。将B*57:01-Tg与PD-1-/-小鼠交配，产生免疫缺陷B*57:01-Tg/PD-1-/-小鼠。CpG-oligodeoxynucleotides和anti-CD4 mAb可进一步调节B*57:01-Tg/PD-1-/-小鼠的免疫应答。然而，B*57:01-Tg小鼠的免疫调节与flx诱导的肝损伤或免疫激活无关。此外，我们生成了FLX-人血清白蛋白（HSA）偶联物，并表明FLX以时间依赖的方式与HSA共价结合。将FLX-HSA偶联物用于B*57:01-Tg小鼠。利用流式细胞术研究免疫应答，揭示CD8+ T细胞（TEM细胞）中CD44highCD62Llow的表型。通过刺激引流淋巴结的TEM细胞，FLX-HSA偶联物引起HLA-B*57:01的限制性免疫反应。综上所述，单独给药FLX对B*57:01-Tg小鼠未引起肝损伤或免疫激活。免疫系统的敏感性在这一过程中并不起决定性作用。FLX和HSA的结合导致特异性TEM细胞刺激，这表明HLA-B*57:01驱动与CD8+ T细胞更强的相互作用。这些结果表明，携带HLA-B*57:01的患者可能更容易受到FLX和白蛋白结合物的影响，并驱动CD8+ T细胞活化，这可能是FLX诱导肝损伤的重要危险因素。此外，FLXHSA加合物的应用可能是构建FLXHSA诱导的小鼠特异性肝损伤的有效方法。
Flucloxacillin (FLX) induces adverse liver reactions, which has been reported to be related to human leukocyte antigen (HLA)-B*57:01. In a previous study, abacavir-induced hypersensitivity was induced in HLA-B*57:01transgenic mice (B*57:01-Tg), originally constructed by our group (Susukida et al., 2021). In this study, B*57:01-Tg mice were used to reproduce FLX-induced liver injury. However, treatment of B*57:01-Tg mice with FLX alone did not increase serum ALT levels. Immune-deficient B*57:01-Tg/PD-1-/-mice were produced by mating B*57:01-Tg with PD-1-/- mice. The immune response of B*57:01-Tg/PD-1-/- mice was further modulated by co-administration of CpG-oligodeoxynucleotides and anti-CD4 mAb. Nevertheless, immune regulation in B*57:01-Tg mice did not contribute to the onset of FLX-induced liver injury or immune activation. Moreover, we generated an FLX-human serum albumin (HSA) conjugate and showed that FLX covalently bound to HSA in a time-dependent manner. The FLX-HSA conjugate was administered to the B*57:01-Tg mice. The immune response was investigated using flow cytometry, revealing the phenotype of CD44highCD62Llow in CD8+ T cells (TEM cells). Administration of the FLX-HSA conjugate resulted in an HLA-B*57:01 restricted immune response as shown by the stimulation of TEM cells in the draining lymph nodes. In conclusion, administration of FLX alone to B*57:01-Tg mice did not induce liver injury or immune activation. Immune system sensitivity does not play a decisive role in this process. The conjugation of FLX and HSA results in specific TEM cell stimulation, which suggests that HLA-B*57:01 drives a stronger interaction with CD8+ T cells. These results suggest that patients carrying HLA-B*57:01 could be more susceptible to a conjugate of FLX and albumin and drive CD8+ T cell activation, which may be a vital risk factor for FLX-induced liver injury. In addition, the application of the FLXHSA adduct may be an effective method for the construction of FLX-induced idiosyncratic liver injury in mice.