Use of Transcriptional Profiling to Delineate the Initial Response of Mice to Intravaginal Herpes Simplex Virus Type 2 Infection

Abstract

Intravaginal (ivag) infection of mice with herpes simplex virus type 2 (HSV-2) causes genital tissue damage, quickly followed by development of fatal encephalopathy. To delineate initial host responses generated by HSV-2 infection, here oligonucleotide microarrays compared gene expression in vaginal tissue from uninfected mice and mice 1, 2, 3, 4, 5, 6, or 7 days after ivag infection with 10 4 pfu HSV-2. While comparison of mRNA expression in uninfected and HSV-infected vaginal tissue detected few changes during the first 2 days post infection (dpi), there were 156 genes whose expression was first significantly altered 3 dpi that remained significantly modified at all later time points examined. These 156 genes were significantly enriched in canonical pathways associated with interferon (IFN) signaling, activation of IFN elements by intracellular pattern recognition receptors, and antiviral immunity induced by cytosolic RIG-like receptors. Evaluation of this gene set with the National Center for Biotechnology Information Gene and INTERFEROME databases corroborated pathway analysis, as function of most (53%) were linked to IFN-mediated host immunity. In the final set of experiments, ivag administration of the Toll-like receptor 3 agonist polyinosinic: polycytidylic acid (poly I:C) 24 h before ivag HSV-2 infection reduced the incidence of genital pathology and encephalopathy, while these poly I:C-treated mice were subsequently protected from ocular HSV-2 challenge lethal to uninfected controls. The latter results imply that the exuberant antiviral immunity produced in our experimental model is simply formed too late to prevent viral replication and dissemination, and that poly I:C-induced formation of an antiviral state protecting against primary ivag infection also permits development of HSV-specific protective immunity.

Dendritic cell function and pathogen-specific T cell immunity are inhibited in mice administered levonorgestrel prior to intranasal Chlamydia trachomatis infection

“…As an important experimental consideration, antecedent progestin treatment is exploited in numerous animal models of genital infection (including murine Chlamydia models that use intravaginal, trans-cervical, and direct ovarian bursal inoculation) to produce uniform susceptibility to infection8192021. We thus tested our hypothesis in a C. trachomatis respiratory infection model, a model in which uniform infection susceptibility is achieved without antecedent progestin treatment.…”

Medroxyprogesterone acetate and levonorgestrel increase genital mucosal permeability and enhance susceptibility to genital herpes simplex virus type 2 infection

“…Mice were sedated with 0.18Qmg xylazine (Lloyd Laboratories, Shenandoah IA) and 1.8Qmg ketamine hydrochloride (JHP Pharmaceuticals, LLC Rochester MI), and ivag infected with 10 4 plaque-forming units (pfu) of WT HSV-2 333 (provided by Dr. Robert Hendricks, University of Pittsburgh) in 10μL of RPMI. Genital pathology was evaluated daily using a previously described 5-point scale: 0, no pathology; 1, mild vulvar erythema; 2, moderate vulvar erythema; 3, severe vulvar erythema and perineal fur loss; 4, perineal ulceration; 5, extension of perineal ulceration and fur loss to surrounding tissue 52 , with mice euthanatized if scores ≥ 4 or encephalopathic changes identified. Other mice were similarly sedated and ivag infected with 3 x 10 6 pfu of HSV-1q-GPF 23 in 10μL of RPMI, and euthanized 1 h, 12 h, or 24 h later.…”

Norethisterone Enanthate Increases Mouse Susceptibility to Genital Infection with Herpes Simplex Virus Type 2 and HIV Type 1

“…Using methods described previously (24), untreated mice in estrus or mice 1 d after their fifth NET-EN treatment or 5 d after a single DMPA treatment were sedated for ivag inoculation with 10 4 PFUs of HSV-2 333 in 10 ml of RPMI 1640 (HSV-2 was kindly provided by Dr. R. L. Hendricks). HSV-induced pathology was evaluated daily via a 5-point scale: 0, no pathology; 1, mild vulvar erythema; 2, moderate vulvar erythema; 3, severe vulvar erythema; 4, perineal ulceration; and 5, extension of perineal ulceration to surrounding areas (29). Mice were euthanized when pathology scores $3 were detected or if they displayed encephalopathic changes.…”