Microbiota-independent antiviral protection conferred by aminoglycoside antibiotics

Smita Gopinath; Myoungjoo V. Kim; Tasfia Rakib; Patrick W. Wong; Michael van Zandt; Natasha A. Barry; Tsuneyasu Kaisho; Andrew L. Goodman; Akiko Iwasaki

To elucidate the mechanisms underlying neomycin-mediated host resistance against HSV-119 2, we analyzed vaginal gene transcription in neomycin-treated mice prior to infection. We 120 identified over a hundred significantly upregulated genes ( Fig. 2a ; Supplementary Table 121 1). No genes were significantly downregulated in expression. Using Ingenuity pathway 122 analysis, we found that genes in the type I interferon (IFN) pathway were heavily enriched 123 ( Fig. 2b) , with over 30% of the upregulated genes falling within this pathway. We 124 independently confirmed the upregulation of a subset of IFN-stimulated-genes (ISGs) 125 using RT-qPCR (Fig. 2c) . ISG expression was also increased in germ-free mice upon 126 neomycin treatment ( Supplementary Fig. 2 ). Neomycin-induced ISG expression was rapid, 127 as a single treatment was sufficient to significantly increase ISG expression 2-5 fold 128 though not to the levels observed after a week's treatment (8-10 fold) ( Supplementary Fig. 129 3a). Neomycin-mediated ISG induction was also dose-dependent as increased amounts of 130 neomycin correspondingly increased ISG expression ( Supplementary Fig. 3a ). Significant 131 ISG induction was maintained up to 3 days after neomycin treatment and was only partially 132 lost by 1 week post neomycin treatment with 5 of 8 genes still significantly upregulated 133 ( Supplementary Fig. 3c ). Finally, this neomycin-mediated ISG induction was restricted to 134 the site of application, as we observed no upregulation of ISG expression in the lungs of 135 neomycin-treated mice ( Supplementary Fig. 3d ). 136 137