Figure 6. HELP eliminates HSV-1 in tissue culture of human corneas.
a, Flow chart for evaluating antiviral effects of HELP in human corneas. 1.5 μg p24 HELP or 15 μL PBS (Mock) were injected into the corresponding punches derived from the same human cornea. After 24 h, the corneal punches were infected with HSV-1 17syn+ (2×106 PFU/piece). b, Confocal analysis of the distribution of HSV-1 and HELP in the human cornea. GFP is indicative of the presence of HELP and VP5 for HSV-1. Scale bar, 30 μm. c, Percentage of VP5+ cells presented in (b). Data shown as the percentage of mock-treated tissues. n=6 biologically independent samples. P<0.0001. d, qPCR analysis the fold change of HSV-1 genome. n=3 biologically independent samples. P=0.0005. e, Titration of supernatants from human corneal cultures. n=3 biologically independent samples. P=0.0003. f, Western blot analysis of VP5 protein. The experiment was repeated twice with similar results. g, Identification of SNV and indel mutations in the HELP treated corneal punch at WGS level. Valid sequencing data were aligned to Human Genome version 19 (HG19). h, Summary of unique SNV and indel mutations. Data and error bars represent mean ± SEM. Unpaired two-tailed Student’s t-tests.