A Systematic Analysis of Host Factors Reveals a Med23-Interferon-? Regulatory Axis against Herpes Simplex Virus Type 1 Replication

Griffiths, Samantha J.; Koegl, Manfred; Boutell, Chris; Zenner, Helen L.; Crump, Colin M.; Pica, Francesca; Gonzalez, Orland; Friedel, Caroline C.; Barry, Gerald; Martin, Kim; Craigon, Marie H.; Chen, Rui; Kaza, Lakshmi N.; Fossum, Even; Fazakerley, John K.; Efstathiou, Stacey; Volpi, Antonio; Zimmer, Ralf; Ghazal, Peter; Haas, Jürgen

Ethnically Italian subjects with or without a history of recurrent Herpes labialis (HL) gave written voluntary informed consent and were enrolled in this study at the University of Rome Tor Vergata with the approval of the Ethical Committee at the University of Rome Tor Vergata. All subjects were interviewed by medically trained investigators using an appropriate questionnaire and agreed to provide saliva and/or blood samples. Data and blood sample collection was carried out as previously described [51] . A total of 58 healthy immunocompetent individuals (17 men and 41 women) age 22-61 years (overall median age 38.5 yr; male median age 40 yr, range 22-61; female median age 38, range 23-60) participated in the study. None of the patients presented with an active lesion at the time of or in the 3 weeks preceding saliva sample collection. For the purpose of this study, patients were characterized into no recurrence of HL (NR), low recurrence (L; 1-3 HL episodes/yr, with a maximum extension of 1 cm, mild symptoms and healing time ,7 days), high recurrence (H; 4 or more HL episodes/yr, extension of lesions more than 1 cm), very high recurrence (H+; more than 4 HL episodes/yr, extension of lesions .3 cm and/or involving nose or cheek beyond the lip, more severe and long lasting associated symptoms including itch, burning, paresthesias and/or neuralgia, with healing times .7 days and who required antiviral therapy (35) (36) (37) (38) (39) (40) (41) (42) (43) (44) (45) (46) ]). Saliva samples (,3 ml) were obtained from each subject after an overnight fast and after rinsing the mouth twice with water, split into two aliquots and frozen at 220uC. Samples were anonymised and stored with dual code labels before shipping on dry ice to the University of Edinburgh for DNA extraction. Saliva and PBMC samples were thawed and DNA extracted using a QIAamp DNA Blood Mini Kit (Qiagen) as per manufacturer's instructions, quantified using a NanoDrop and IL28B genotype determined by melt-curve analysis PCR on a LightCycler480 (Roche) using the LightMixH Kit IL28B (TIB Molbiol) as per manufacturer's instructions. Significance of genotype association was determined by Fisher's exact test, comparing the frequency of the CC, CT or TT genotype in the NR group versus the L (pvalue = 1), H (p-value = 0.12) or H+ (p-value = 0.015) clinical groups.