HSV-infected Mice Research Articles

Topical tenofovir, a microbicide effective against HIV, inhibits herpes simplex virus-2 replication.

The HIV reverse-transcriptase inhibitor, tenofovir, was recently formulated into a vaginal gel for use as a microbicide. In human trials, a 1% tenofovir gel inhibited HIV sexual transmission by 39% and, surprisingly, herpes simplex virus-2 (HSV-2) transmission by 51%. We demonstrate that the concentration achieved intravaginally with a 1% tenofovir topical gel has direct antiherpetic activity. Tenofovir inhibits the replication of HSV clinical isolates in human embryonic fibroblasts, keratinocytes, and organotypic epithelial 3D rafts, decreases HSV replication in human lymphoid and cervicovaginal tissues ex vivo, and delays HSV-induced lesions and death in topically treated HSV-infected mice. The active tenofovir metabolite inhibits HSV DNA-polymerase and HIV reverse-transcriptase. To exert dual antiviral effects, tenofovir requires topical administration to achieve a drug concentration higher than systemic levels achieved by oral treatment. These findings indicate that a single topical treatment, like tenofovir, can inhibit the transmission of HIV and its copathogens.

Activation of PKR Causes Amyloid ß-Peptide Accumulation via De-Repression of BACE1 Expression

BACE1 is a key enzyme involved in the production of amyloid ß-peptide (Aß) in Alzheimer's disease (AD) brains. Normally, its expression is constitutively inhibited due to the presence of the 5′untranslated region (5′UTR) in the BACE1 promoter. BACE1 expression is activated by phosphorylation of the eukaryotic initiation factor (eIF)2-alpha, which reverses the inhibitory effect exerted by BACE1 5′UTR. There are four kinases associated with different types of stress that could phosphorylate eIF2-alpha. Here we focus on the double-stranded (ds) RNA-activated protein kinase (PKR). PKR is activated during viral infection, including that of herpes simplex virus type 1 (HSV1), a virus suggested to be implicated in the development of AD, acting when present in brains of carriers of the type 4 allele of the apolipoprotein E gene. HSV1 is a dsDNA virus but it has genes on both strands of the genome, and from these genes complementary RNA molecules are transcribed. These could activate BACE1 expression by the PKR pathway. Here we demonstrate in HSV1-infected neuroblastoma cells, and in peripheral nervous tissue from HSV1-infected mice, that HSV1 activates PKR. Cloning BACE1 5′UTR upstream of a luciferase (luc) gene confirmed its inhibitory effect, which can be prevented by salubrinal, an inhibitor of the eIF2-alpha phosphatase PP1c. Treatment with the dsRNA analog poly (I∶C) mimicked the stimulatory effect exerted by salubrinal over BACE1 translation in the 5′UTR-luc construct and increased Aß production in HEK-APPsw cells. Summarizing, our data suggest that PKR activated in brain by HSV1 could play an important role in the development of AD.

Topical cream-based oxyresveratrol in the treatment of cutaneous HSV-1 infection in mice

Anti-herpes simplex virus (HSV) activities of oxyresveratrol in vitro and topical administration in cutaneous HSV-1 infection in mice were examined. The inhibitory concentrations for 50% plaque formation (IC 50) of oxyresveratrol against HSV-1 clinical isolates and HSV-2 clinical isolates were 20.9–29.5 and 22.2–27.5 μg/ml, respectively. In topical administration in cutaneous HSV-1 infection in mice, 2.5%, 5%, 10% and 20% oxyresveratrol in cream vehicle applied three times daily for 7 days after infection were evaluated and 10% and 20% oxyresveratrol cream were significantly effective in delaying the development of skin lesions and protection from death ( P < 0.01). The concentration of 10% oxyresveratrol in cream was significantly more effective than that of 30% oxyresveratrol in vaseline applied three times daily ( P < 0.01). Oxyresveratrol cream at 20% was as effective as 5% ACV cream applied three times daily ( P < 0.01). Both 10% and 20% oxyresveratrol cream were as effective as that of 5% ACV cream applied two times daily ( P > 0.05). Therapeutic efficacy of oxyresveratrol in cream vehicle was dose-dependent and the maximum efficacy observed on day 6 after infection was shown at 10% oxyresveratrol in cream applied three times daily. The frequency of application of 10% oxyresveratrol cream at three, four and five times daily was as effective as that of 5% ACV cream applied five times daily ( P > 0.05). These results demonstrated that topical administration of oxyresveratrol in novel cream vehicle reduced the concentration of oxyresveratrol to 10% and was suitable for cutaneous HSV infection.

Effect of LIGHT adjuvant on kinetics of T-cell responses induced by HSV-1 DNA immunization.

Studies on efficacy of various vaccines that prevent or reduce the primary and recurrent HSV-1 infection have demonstrated the importance of cellular immunity for protection against the infection. We previously used DNA vaccination to induce cellular immunity against HSV-1 infection in mice. The aim of our study was to evaluate the effect of LIGHT; a member of TNF super family, on the kinetic of CTL response induced by HSV-1 glycoprotein B based DNA vaccine. Using a granzyme B ELISA for detection and analysis of CD8+ T cells, CTL activity was determined in the spleen of BALB/c mice at various time points after primary and booster dose of vaccination. The kinetics of CTL response to primary and secondary HSV-1 infection and DNA vaccination were compared to those induced by DNA vaccination in combination with LIGHT adjuvant in the present study. In primary and secondary immunization, the CTL activity in the HSV injected group peaked 7 days and 12 hours post immunization, respectively. After 5 days, LIGHT could neither accelerate the CTL response compared to DNA vaccination alone nor could enhance the CTL activity in the primary and the first peak of memory response, the amount of granzyme B induced by the LIGHT containing vaccine was significantly higher than that induced by the vaccine without the adjuvant. Although LIGHT enhances the cellular response in the booster dose of vaccination, it does not accelerate the CTL response.

Controlling viral immunoinflammatory lesions by modulating aryl hydrocarbon receptor signaling (168.19)

Abstract Ocular HSV-1 infection can cause a blinding CD4+ T cell orchestrated immunoinflammatory lesion called Stromal Keratitis (SK). Lesion severity is affected by the balance of proinflammatory effector T and regulatory T cells (Treg). In this report we show that expanding Treg activity using TCDD (2, 3, 7, 8- Tetrachlorodibenzo-p-dioxin), a ligand for the AhR receptor, results in the differential expansion of Treg in the eye and a resultant reduction in the extent of inflammatory lesions. Treatment of HSV infected mice with TCDD either on day 1 or day 5 pi, diminished the severity of SK lesions as well as the extent of corneal angiogenesis. The observed effect was mainly due to the changed balance of CD4+ T cells in the cornea that were IFN-γ or IL17 cytokine producers compared to those that were Foxp3+ Tregs. The change was explained by the significantly increased apoptosis of activated effector T cells but not of Tregs. Additionally, effector T cells from HSV infected mice proliferated significantly less in TCDD treated mice compared to control mice. Furthermore the addition of TCDD to the culture of antiCD3/CD28 stimulated naïve CD4+ T cells caused significant induction of Tregs. In conclusion, our results demonstrate that modulation of AhR signaling by TCDD represents a valuable approach to expand Treg and inhibit Th1 and Th17 cells to control HSV induced blinding lesions.

An influence of galectin-9/Tim-3 interaction on Herpes simplex virus-1 latency (105.14)

Abstract In this report, we show that the majority of CD8+T cells in the trigeminal ganglion (TG) of mice ocularly infected with HSV-1 express Tim-3, a molecule that delivers negative signals to CD8+T cells when it engages its ligand, Galectin 9 (Gal-9). Curiously Gal-9 glycoprotein is also up regulated in the TG both during the time when replicating virus is present as well during early viral latency. Accordingly it is conceivable that the efficiency of ganglionic CD8+T cell function could be influenced by Gal-9/Tim-3 interaction. In support of this we show that the activity of peptide stimulated CD8+T cells recovered from the TG of WT mice produced less IFN-γ, TNF-α and Granzyme B molecules in comparison to TG CD8+T cells of Gal-9KO mice. Differences in the number of productively and latently infected neuronal cells were also evident between WT and Gal-9KO mice. In vitro studies demonstrated that TG cell cultures of WT HSV-1 infected mice exposed to recombinant Gal-9 in the latent phase caused apoptosis of some CD8+T cells and induce viral reactivation. Our results demonstrate that the host homeostatic mechanism mediated by Gal-9/Tim-3 interaction on CD8+T cells can influence the outcome of latent HSV-1 infection. Neuronal cell function is likely to be preserved when host Gal-9 levels are low, since inhibitory effects on latency controlling CD8+T cells will be minimal.

The effect of emodin, an anthraquinone derivative extracted from the roots of Rheum tanguticum, against herpes simplex virus in vitro and in vivo

Herpes simplex viruses (HSV-1 and -2) are important pathogens for humans and the discovery of novel anti-HSV drugs with low toxicity deserves great efforts. Rhubarb is one of the oldest and best-known traditional Chinese medicines. We initiated this study to test if emodin is the active ingredients from Rheum tanguticum (R. tanguticum, one of the Chinese Rhubarb) against HSV infection and to investigate its antiviral activity on HSV infection in tissue culture cells and in a mouse model. Emodin (3-methyl-1,6,8-trihydroxyanthraquinone) was extracted and purified from R. tanguticum (cultivated at high mountainous area in Qinghai) and the purity was determined by high performance liquid chromatography. The antiviral experiments of emodin against HSV infection were performed in vitro and in vivo. In vivo, the HSV-infected mice were orally administered with emodin beginning at 24 h post-HSV exposures with dosages of 3.3 g/kg/day, 6.7 g/kg/day, and 11.3 g/kg/day, respectively, for 7 days. Emodin was found to inhibit the replication of HSV-1 and HSV-2 in cell culture at the concentration of 50 μg/ml with antiviral index of 2.07 and 3.53, respectively. The emodin treatment increased the survival rate of HSV-infected mice, prolonged survival time and showed higher efficacy of HSV elimination from brain, heart, liver and ganglion, compared to the viral controls. In addition, the antiviral activity of emodin was found to be equivalent to that of acyclovir in vivo. Our results indicate that emodin has the anti-HSV activity in vitro and in vivo and is thus a promising agent in the clinical therapy of HSV infection.

An Imbalance in sVEGFR1 and VEGF-A mediates corneal neovascularization after HSV-1 ocular infection (137.23)

Abstract Ocular transparency is essential for optimal vision. The cornea maintains avascularity by expressesing soluble VEGFR1 (sVR) which binds resident VEGFA and constrain its pro-angiogenic activity. Here we report that early on, herpes simplex virus 1 (HSV-1) infection leads to an increase in VEGFA expression while low sVR levels in vitro as well as in vivo in infected mice corneas. In addition, we found degradation of sVR protein which was subsequently attributed to matrix metallopeptidase 7 and 9 as observed by in-vitro and ex-vivo degradation assays. Furthermore, mice receiving an inhibitor of MMPs showed decreased degradation of sVR and diminished HSV induced corneal angiogenesis. The second peak of VEGFA observed during clinical phase of herpes stromal keratitis (HSK) was attributed to infiltrating neutrophils and their depletion during this stage resulted in low VEGFA load and diminished corneal angiogenesis. Finally, the provision of recombinant sVR to HSV infected mice diminished angiogenic response supporting the notion that the activity of sVR has a constraining effect on the angiogenic activity of VEGFA. Taken together our results support the notion that the binding of sVR to VEGFA acts to limit the extent of ocular angiogenesis in pathological situations such as observed in HSK. Thus therapies designed to increase the presence, or diminish the breakdown, of sVR may be a useful therapeutic approach to manage the severity of HSK, an important cause of human blindness.

Effect of Corilagin on anti-inflammation in HSV-1 encephalitis and HSV-1 infected microglias

The aim of this explore is to study the anti-inflammatory effect of Corilagin in herpes simplex virus (HSV)-1 infected microglial cells and HSV-1 infected mouse brain. The cellular model was set with microglial cells stimulated by HSV-1 and divided respectively, into virus, astragalus polysaccharides (APS), Dexamethasone and Corilagin group. A normal control group consisting of uninfected microglial cells was also included. ELISA for measuring TNF-α, IL-1β and IL-10 and Greiss method for detecting NO secretion in supernatant, flow cytometry assay for examining apoptosis rate, expression of caspase-3, caspase-8, caspase-9 and caspase-12, and western-blot for measuring protein expression of cytochrome c were performed. The animal model was set up using Balb/c male mice that were intracranially inoculated with HSV-1. Animals were then divided in groups as described for the cellular model. Here, too a normal control group was included. HE staining was used to assay pathological changes in brain. As results, after Corilagin intervention, the release of TNF-α, IL-1β and NO from HSV-stimulated migroglia cells was significantly inhibited. Furthermore, Corilagin induced apoptosis of HSV-stimulated microglia through all the 3 known apoptotic pathways. The animal model treated with Corilagin also displayed significant decrease of herpes simplex encephalitis induced brain pathological changes. In conclusion, Corilagin has the potential to reduce HSV-1-induced inflammatory insult to the brain, and its mode of action is through the induction of apoptosis of microglias and reduction of cytokines production.

Expression of Herpes Virus Entry Mediator (HVEM) in the Cornea and Trigeminal Ganglia of Normal and HSV-1 Infected Mice

Purpose: Herpes virus entry mediator (HVEM) plays a critical role in the regulation of inflammation through interaction with its natural ligands LIGHT and lymphotoxin alpha and also serves as one of the entry receptors of herpes simplex virus (HSV). The purpose of this study was to better understand the expression of HVEM in the cornea and trigeminal ganglia (TG), which are important targets of HSV infection.Materials and Methods: Immunohistochemistry was used to define HVEM expression in the cornea and TG of normal and HSV-1 infected mice euthanized 2 to 5 days or 7 months following corneal inoculation of virus.Results: We found that HVEM is widely expressed in the normal corneal epithelium and endothelium, is weakly and focally expressed in the corneal stroma, and is expressed in a portion of neurons and non-neuronal cells in the TG. Acute HSV-1 keratitis and ganglionitis were associated with increased HVEM expression in the corneal epithelium and stroma and in neurons and non-neuronal cells of TG, and many inflammatory cells in these tissues also expressed HVEM. TG derived from mice 7 months after virus inoculation demonstrated latent HSV-1 infection that was associated with increased HVEM expression in neurons and non-neuronal cells relative to uninfected control tissues. Latent TG also contained focal infiltrates of mononuclear inflammatory cells, many of which expressed HVEM. Corneas derived from latently infected mice demonstrated chronic keratitis, with no evidence of virus replication or increased HVEM expression in the corneal epithelium, and inflammatory cells present showed only weak HVEM expression.Conclusions: HVEM is expressed in the cornea and TG and therefore may serve as an HSV entry receptor in these tissues. Furthermore, these findings raise the possibility that changes in HVEM expression following ocular HSV-1 infection can modulate HSV spread and infection-induced inflammation in the cornea and TG.

Inhibition of HSV-1 ocular infection with morpholino oligomers targeting ICP0 and ICP27

Alternative therapies are needed for HSV-1 infections in patients refractory to treatment with Acyclovir (ACV) and its derivatives. Peptide-conjugated phosphorodiamidate morpholino oligomers (PPMO) are single-stranded DNA analogues that enter cells readily and reduce target gene expression through steric blockage of complementary RNA. When applied before or soon after infection PPMO targeting the translation-start-site regions of HSV-1 ICP0 or ICP27 mRNA reduced HSV-1 plaque formation by 70–98% in vitro. The ICP0 PPMO also reduced ACV-resistant HSV-1 (strain 615.9) plaque formation by 70–90%, while an equivalent dose of ACV produced only 40–50% inhibition when the treatment was applied between 1 and 3hpi. Seven daily topical treatments of 100μg ICP0 PPMO caused no gross or microscopic damage to the corneas of uninfected mice. Topical application of 10μg ICP0 PPMO to the eyes of HSV-1 infected mice reduced the incidence of eye disease by 37.5–50% compared to controls. This study demonstrates that topically applied PPMO holds promise as an antiviral drug candidate against HSV-1 ocular infection.

Psychological stress impairs the local CD8 + T cell response to mucosal HSV-1 infection and allows for increased pathogenicity via a glucocorticoid receptor-mediated mechanism

Psychological stress and its associated increases in corticosterone are generally immunosuppressive and contribute to increased herpes simplex virus (HSV)-associated pathogenicity. However, the impact of stress on local control of the initial mucosal-based HSV infection has not been elucidated, nor have the ramifications of such failures of the immune response in terms of viral spread. To address these gaps in knowledge, the studies described herein sought to determine how psychological stress and associated increases in corticosterone may increase susceptibility to HSV encephalitis by allowing for increased viral titers at the site of initial infection. We have shown that in mice intranasally infected with HSV-1, a cell-mediated immune response occurs in the nasopharyngeal-associated lymphoid tissue (NALT), mediastinal lymph nodes (MLN), and superficial cervical lymph nodes (CLN). However, psychological stress induced by restraint decreased the number of lymphocytes in these tissues in HSV-infected mice. Surprisingly, the effects of this restraint stress on HSV-specific CTL function varied by immune tissue. Increased viral titers were found in the nasal cavity of stressed mice, an observation which correlated with an increased CD8+ cell response in the CLN. These findings led us to extend our studies to also determine the ramifications of decreased numbers of locally derived lymphocytes on viral titers following infection. Using an approach in which the NALT was surgically removed prior to infection, we confirmed that decreased numbers of NALT-derived lymphocytes at the time of infection allows for increased viral replication. We conclude that the increased viral titers observed in mice experiencing psychological stress are the consequence of a glucocorticoid-mediated reduction in the numbers of lymphocytes responsible for resolving the initial infection.

Anti-herpes simplex virus (HSV-1) activity of oxyresveratrol derived from Thai medicinal plant: Mechanism of action and therapeutic efficacy on cutaneous HSV-1 infection in mice

Oxyresveratrol, a major compound purified from Artocarpus lakoocha, a Thai traditional medicinal plant, was evaluated for its mechanism of action and therapeutic efficacy on cutaneous herpes simplex virus (HSV) infection in mice. The inhibitory concentrations for 50% HSV-1 plaque formation of oxyresveratrol, three clinical isolates, thymidine kinase (TK)-deficient and phosphonoacetic acid (PAA)-resistant HSV-1 were 19.8, 23.3, 23.5, 24.8, 25.5 and 21.7 μg/ml, respectively. Oxyresveratrol exhibited the inhibitory activity at the early and late phase of viral replication and inhibited the viral replication with pretreatment in one-step growth assay of HSV-1 and HSV-2. Oxyresveratrol inhibited late protein synthesis at 30 μg/ml. The combination of oxyresveratrol and acyclovir (ACV) produced synergistic anti-HSV-1 effect, as characterized by the isobologram of plaque inhibition. Mice orally treated with oxyresveratrol (500 mg/kg/dose) dose at 8 h before and three times daily had significant delay in herpetic skin lesion development ( P < 0.05). Topical application of 30% oxyresveratrol ointment five times daily significantly delayed the development of skin lesions and protected mice from death ( P < 0.0001).

IL-10 and Natural Regulatory T Cells: Two Independent Anti-Inflammatory Mechanisms in Herpes Simplex Virus-Induced Ocular Immunopathology

Two prominent anti-inflammatory mechanisms involved in controlling HSV-1-induced corneal immunopathology (stromal keratitis or SK) are the production of the cytokine IL-10 and the activity of natural regulatory T cells (nTregs). It is not known whether, under in vivo conditions, IL-10 and nTregs influence the corneal pathology independently or in concert. In the current study using wild-type and IL-10(-/-) animals, we have assessed the activity of nTregs in the absence of IL-10 both under in vitro and in vivo conditions. The IL-10(-/-) animals depleted of nTregs before ocular infection showed more severe SK lesions as compared with the undepleted IL-10(-/-) animals. In addition, nTregs purified from naive WT and IL-10(-/-) animals were equally able to suppress the proliferation and the cytokine production from anti-CD3-stimulated CD4(+)CD25(-) T cells in vitro. Furthermore, intracellular cytokine staining results indicated that nonregulatory cells expressing B220 and CD25 markers were the major IL-10-producing cell types in the lymphoid tissues of HSV-infected mice. In contrast, in the infected corneas, cells with the CD11b(+)Gr1(+) phenotype along with a minor population of Foxp3(-)CD4(+) and a few F4/80(+) cells produced IL-10. Our current investigations indicate that at least two independent anti-inflammatory mechanisms are involved in limiting the corneal lesions in SK, both of which may need to be modulated to control SK therapeutically.

Experimental study of suppression of reactivation of herpes simplex virus type 1 by cyclooxygenase 2 inhibitor with acyclovir

To study whether the cyclooxygenase 2 (COX-2) inhibitor can block the herpes virus reactivation and whether the combination of COX-2 inhibitor with acyclovir can enhance the inhibition of virus reactivation. It was a experimental study. Mice were randomly divided into six groups. Five groups were HSV-1 infected mice, which included: group A, treated with lornoxicam and acyclovir; groups B and C, treated with lornoxicam or acyclovir, respectively; groups E and F were injected with saline as the untreated control groups. The sixth group was uninfected mice as the control group. All groups were undergone to reactivate the herpes virus by UV-B except group F. The shedding of the virus was determined by cultures of ocular swab or ganglion homogenates with indicator cells. The rates of corneas and ganglia containing the infectious virus in the groups A, B and C were significantly lower than those in the control group D, (cornea: 2XA-D = 36.88, XB-D = 22.43, X2C-D = 20.32, P < 0.05, ganglia : X2A-D = 49.91 X2B-D =29. 16,X2C.D = 24.89, P < 0.05). Combined use of these two drugs in group A showed no significant statistical difference as compared with using them separately in the cornea culture (X2A-B= 2.75, X2A-C = 3.66, 0. 05 < P < 0.1), but there was significant difference in trigeminal ganglia culture (X2A-B = 4.78, 2XA-c = 6. 97, P < 0.05). These experiments demonstrate that a selective COX-2 inhibitor can suppress UV-B-induced herpes virus reactivation in the cornea and nervous system. A combination of acyclovir does not significantly enhance the inhibition of virus reactivation by lornoxicam. These results provide a new method to prevent the recurrence of HSK.

Neonatal CD8+ T cells are slow to develop into lytic effectors after HSV infection in vivo

HSV is an important neonatal pathogen. We defined the kinetics of the primary CTL response to HSV-2 in vivo in neonatal mice. Using a replication-defective HSV-2 virus, we demonstrate that neonates mount a primary HSV-specific CTL effector response in the draining LN, with delayed onset and shortened peak activity, in contrast to the rapid, strong response observed in adult mice. The shortened peak neonatal CTL response is independent of HSV dose and is associated with retarded CD8(+) T cell expansion, reduced expansion of HSV-specific tetramer-positive CD8(+) T cells and a reduced CD8(+) T cell IFN-gamma response. Paradoxically, neonatal CD8(+) T cells display enhanced non-specific early activation that is not sustained. Neonatal HSV-specific TCR-transgenic CD8(+) T cells showed reduced proliferation in vivo when transferred into HSV-infected neonatal mice compared to adult T cell controls. Our data suggest that early events in CD8(+) T cell priming underlie the attenuated newborn CTL response to HSV.

Effects of CXCR3 Signaling on Development of Fatal Encephalitis and Corneal and Periocular Skin Disease in HSV-Infected Mice Are Mouse-Strain Dependent

The host inflammatory response to ocular infection with herpes simplex virus (HSV) can be either protective, with disease-free survival, or it can promote diseases such as HSV corneal disease (or herpes stromal keratitis [HSK] in humans) and encephalitis (HSE), depending on mouse strain. The role of CXCR3 chemokine signaling in HSV-induced central nervous system (CNS) inflammation and corneal disease was evaluated, and responses in genetically susceptible and resistant strains of mice were contrasted. Resistant C57BL/6J (B6) and susceptible 129S6 (129) mice were given monoclonal antibodies (mAbs) to neutralize the CXCR3 ligands monokine induced by interferon-gamma (MIG, CXCL9) and interferon inducible protein-10 (IP-10, CXCL10) during HSV infection. In addition, the development of HSV disease was monitored in CXCR3-null mutant mice derived from resistant (B6) and susceptible (BALB/c) strains. Inflammatory cells infiltrating the cornea and brain stem were isolated and stained for flow cytometric analysis. MIG and IP-10 were induced in nervous system tissue after HSV inoculation by the corneal route. HSV-infected 129 mice treated with MIG- or IP-10-neutralizing mAbs showed significantly enhanced survival compared with mice treated with control isotype antibody, whereas survival of the B6 mice was unaltered. Similarly, greater survival was observed for BALB.CXCR3(-/-) mice compared with control BALB/c mice. Reduced CNS inflammation was documented that extended to the cornea, such that HSV corneal disease severity was reduced in susceptible BALB.CXCR3(-/-). In contrast, although survival of B6 and B6.CXCR3(-/-) mice was indistinguishable, B6.CXCR3(-/-) mice developed more severe corneal and periocular skin disease. The effects of CXCR3 signaling in HSV infection are strongly dependent on mouse strain.

Evaluation of γ-interferon kinetics in HSV-1 infected mice in different days post infection (in vivo) and post re-stimulation (in vitro)

Gamma interferon (IFN-gamma) is among the most important immune factors for limiting of herpes simplex virus (HSV) infections. However, our knowledge about the kinetics of IFN-gamma production after HSV infection is limited. The present study examines the kinetics of IFN-gamma expression following secondary infection with HSV-1. Using semiquantitative RT-PCR assay, the expression of IFN-gamma in spleen lymphocytes was significantly detected on 14 days but not 7 days after intraperitoneal inoculation of HSV-1, while ELISA detected IFN-gamma on both days. At various hours after in vitro re-stimulation of spleen cells, RT-PCR showed a decreasing pattern of mRNA transcripts, whereas, ELISA assayed an increasing amount of secreted protein through the experiment. Despite the contrast results of ELISA and RT-PCR, regarding the short half-life of mRNA, the data are in correlation with each other and need to interpret.

Effect of Orally Administered Oat ?-Glucan on the Macrophage Response to HSV-1 Infection Following Exercise Stress

Oat β-glucan (OβG), a soluble fiber with mild immunostimulant activity, can offset the increased risk of infection associated with exercise stress (Davis et. al., Med Sci Sports Exerc 36(8): 1321 −7, 2004). Macrophages (Mθs) play an important role in this mechanism: we have shown that depletion of Mθs negates the beneficial effect of OβG on preventing the exercise induced increase in susceptibility to infection (Murphy et. al., Med Sci Sports Exerc 37 (5); S128, 2005). Specifically, we have found that OβG can offset the decrease in intrinsic anti-viral resistance of Mθs associated with stressful exercise, however it is likely that extrinsic functions of Mθs are also involved. PURPOSE: To examine the effect of oral feedings of OβG on Mβ cytokine release following exercise stress and in vitro HSV-1 infection in mice. METHODS: Exercise mice (Ex-H2O & Ex-OβG) were run to fatigue on a treadmill for 3 consecutive days, control mice (Con-H2O & Con-OβG) remained in their cages in the treadmill room. OβG was fed to the mice in their drinking water for 10 consecutive days. Mice were sacrificed 15 minutes following the final exercise bout or rest. Lung and peritoneal Mθs were harvested via lavage, and allowed to adhere on a 96 well microplate at a concentration of 1 X 106. A standardized dose of HSV-1 was incubated with the Mθs for a period of 24h after which time supernatants were collected. Mβ supernatants were analyzed for IL-1β, IL-6 and TNF-α via ELISA. RESULTS: Both exercise and OβG increased IL-6 and TNF-α release from peritoneal and lung Mθs following HSV-1 infection. OβG increased IL-1β release from peritoneal Mθs following infection. RESULTS: indicate that both exercise and oral feedings OβG can increase the Mθ cytokine response to HSV-1 in vitro. CONCLUSION: The benefits of OβG on preventing the exercise induced increase in susceptibility to infection may be mediated at least in part by certain extrinsic functions of Mθs. This study was funded by a student research grant from the American College of Sports Medicine.

Acute morphine exposure potentiates the development of HSV-1-induced encephalitis

A devastating consequence of HSV-1 infection is development of HSV-1-induced encephalitis (HSVE). While only a minority of individuals infected with HSV-1 experiences HSVE, clearly defined variables that consistently predict development of the disease remain to be elucidated. The current study examined the effects of a single dose of morphine prior to infection with HSV-1 on the development of HSVE in BALB/cByJ mice. Acute morphine exposure was observed to potentiate the development of HSVE in HSV-1 infected mice. The present data implicate a potential role for the blood–brain barrier in the development of HSVE in morphine-treated mice.