Differences in Nature Killer cell Response and Interference with Mitochondrial DNA induced Apoptosis in Moxifloxacin Environment

In light of the widespread adoption of antibiotics, the imperative to promptly manage and regulate their use, ensuring accuracy and safety, is evident. Such misuse can lead to frequent adverse drug reactions, causing unnecessary harm, and prolonged erroneous use may culminate in irreversible injuries and fatalities. Of particular concern is the prevalent use of moxifloxacin (MXF), a fourth-generation fluoroquinolone antibiotic, across various infections and disease prevention scenarios. This have been reported to have immunomodulatory effects on a variety of immune cells and even anti-proliferative and pro-apoptotic effects, but the mechanism of action is not fully clear. There exists an urgent necessity to investigate the safety and mechanisms of action on immunomodulation of this drugs. Overall design: The study involved the in vitro treatment of peripheral blood mononuclear cells (PBMC) from healthy adults (n=3) with MXF (10ug/ml) for 24 hours. Single-cell sequencing was employed to delineate the impacts of MXF on various immune cell taxa both before and after its administration. Subsequent to MXF exposure, the identification of subsets displaying the most significant damage in Natural Killer (NK) cells was accomplished, followed by the screening of pivotal differential genes. Phenotyping of the most affected NK cell subsets was executed using flow cytometry. The exploration of drivers influencing NK cell differentiation stimulated by MXF was achieved through mimetic time-series analysis. The differential genes extensively down-regulated in NK cells were localised within the mitochondrial respiratory chain complexes. The endpoints of MXF-induced stress damage in NK cells were identified through assays measuring mitochondrial respiratory chain complexes I, III, IV, and ATP synthase activity. Additionally, assessments included mitochondrial membrane potential, reactive oxygen species, and mitochondrial electron microscopy.