2026-10-14T04:39:00Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34036181 2026-09-30T17:44:53Z category_7 category_12 category_21 category_46 category_64 category_132 category_134 portal_5 item_type_1 month_year_09_2026

Upstream Variability in IFNγ Signaling Amongst Keratinocytes from Asymptomatic Donors and Donors with Ulcerative Disease. Angelina Zimenko (25145435) Ian Hayman (25145438) Youyi Fong (519951) Luke D. Monroe (5165948) Margaret G. Mills (3756298) Alex Greninger (11084570) Christine Johnston (375038) Lawrence Corey (140058) Jia Zhu (135506) Medicine Cell Biology Biotechnology Immunology Cancer Infectious Diseases Virology strongly induced gene protein levels compared limited genital lesions infects genital epithelia increased nuclear localization including robust induction ifn &# 947 enhanced chemokine production immune cell recruitment epithelial antiviral defense antiviral immune cells asymptomatic viral shedding doses much lower cell cytokine signaling epithelial cells efficient recruitment either asymptomatic wide spectrum vaccine strategies understanding variability symptomatic donors results suggest results revealed recurrent self previously reported interferon gamma improving therapies clinical outcomes chemokines involved Keratinocytes isolated from asymptomatic donors (A) or donors with ulcerative disease (B) were pretreated with specified doses of IFNγ for 24 hours, then infected with GFP-expressing HSV-1. Live cell images were acquired every 2 hours for 48 hours and normalized to measure GFP fluorescence. (C) The amount of IFNγ required to inhibit 50% of viral GFP expression was calculated for keratinocytes from both asymptomatic and ulcerative donors at 24 hours post infection. (D) Expression of IFNGR1 and STAT1 was measured by RT-qPCR in untreated keratinocytes from asymptomatic or ulcerative donors. (E) Representative images from a single donor illustrating the detection of pSTAT1 in the nucleus of keratinocytes treated with various doses of IFNγ for the specified time. (F) Representative images from each donor at 60 minutes following treatment with 1U of IFNγ highlight heterogeneity in nuclear pSTAT1 signal. Yellow is nuclear DAPI staining, blue is membrane staining with wheat-germ agglutinin, and pink is pSTAT1. Scale bar represents 100 µm (E and F). (G) Normalized average nuclear fluorescence of pSTAT1 at specified times following treatment with specified doses of IFNγ, separated by donor cohort. Data was normalized using the asymptomatic donor pool (0% set by 0U IFNγ, 100% set by 100U IFNγ). Each point is derived from the average fluorescence of at least 29,000 nuclei. (H) Box-and-whisker plot of raw nuclear fluorescence of pSTAT1 at 240 minutes following treatment with specified doses of IFNγ, separated by donor severity. Each bar represents the inner quartile range of raw fluorescence in at least 29,000 nuclei. *P < 0.05, ** P < 0.01, *** P < 0.001, NS, not significant; Unpaired t-test models were used to analyze fold change in gene expression. N = 4 donors and 3 replicates per donor in each cohort (A-D) or 5 donors and 2 replicates per donor in each cohort (E-H). 2026-09-30T17:55:35Z Image Figure 10.1371/journal.ppat.1014612.g006 https://figshare.com/articles/figure/_p_Upstream_Variability_in_IFN_Signaling_Amongst_Keratinocytes_from_Asymptomatic_Donors_and_Donors_with_Ulcerative_Disease_p_/34036181 CC BY 4.0