Neurotropic Mouse Hepatitis Virus Research Articles

CD40L protects against mouse hepatitis virus-induced neuroinflammatory demyelination

Neurotropic mouse hepatitis virus (MHV-A59/RSA59) infection in mice induces acute neuroinflammation due to direct neural cell dystrophy, which proceeds with demyelination with or without axonal loss, the pathological hallmarks of human neurological disease, Multiple sclerosis (MS). Recent studies in the RSA59-induced neuroinflammation model of MS showed a protective role of CNS-infiltrating CD4+ T cells compared to their pathogenic role in the autoimmune model. The current study further investigated the molecular nexus between CD4+ T cell-expressed CD40Ligand and microglia/macrophage-expressed CD40 using CD40L-/- mice. Results demonstrate CD40L expression in the CNS is modulated upon RSA59 infection. We show evidence that CD40L-/- mice are more susceptible to RSA59 induced disease due to reduced microglia/macrophage activation and significantly dampened effector CD4+ T recruitment to the CNS on day 10 p.i. Additionally, CD40L-/- mice exhibited severe demyelination mediated by phagocytic microglia/macrophages, axonal loss, and persistent poliomyelitis during chronic infection, indicating CD40-CD40L as host-protective against RSA59-induced demyelination. This suggests a novel target in designing prophylaxis for virus-induced demyelination and axonal degeneration, in contrast to immunosuppression which holds only for autoimmune mechanisms of inflammatory demyelination.

Neuronal Ablation of Alpha/Beta Interferon (IFN-α/β) Signaling Exacerbates Central Nervous System Viral Dissemination and Impairs IFN-γ Responsiveness in Microglia/Macrophages.

Alpha/beta interferon (IFN-α/β) signaling through the IFN-α/β receptor (IFNAR) is essential to limit virus dissemination throughout the central nervous system (CNS) following many neurotropic virus infections. However, the distinct expression patterns of factors associated with the IFN-α/β pathway in different CNS resident cell populations implicate complex cooperative pathways in IFN-α/β induction and responsiveness. Here we show that mice devoid of IFNAR1 signaling in calcium/calmodulin-dependent protein kinase II alpha (CaMKIIα) expressing neurons (CaMKIIcre:IFNARfl/fl mice) infected with a mildly pathogenic neurotropic coronavirus (mouse hepatitis virus A59 strain [MHV-A59]) developed severe encephalomyelitis with hind-limb paralysis and succumbed within 7 days. Increased virus spread in CaMKIIcre:IFNARfl/fl mice compared to IFNARfl/fl mice affected neurons not only in the forebrain but also in the mid-hind brain and spinal cords but excluded the cerebellum. Infection was also increased in glia. The lack of viral control in CaMKIIcre:IFNARfl/fl relative to control mice coincided with sustained Cxcl1 and Ccl2 mRNAs but a decrease in mRNA levels of IFNα/β pathway genes as well as Il6, Tnf, and Il1β between days 4 and 6 postinfection (p.i.). T cell accumulation and IFN-γ production, an essential component of virus control, were not altered. However, IFN-γ responsiveness was impaired in microglia/macrophages irrespective of similar pSTAT1 nuclear translocation as in infected controls. The results reveal how perturbation of IFN-α/β signaling in neurons can worsen disease course and disrupt complex interactions between the IFN-α/β and IFN-γ pathways in achieving optimal antiviral responses.IMPORTANCE IFN-α/β induction limits CNS viral spread by establishing an antiviral state, but also promotes blood brain barrier integrity, adaptive immunity, and activation of microglia/macrophages. However, the extent to which glial or neuronal signaling contributes to these diverse IFN-α/β functions is poorly understood. Using a neurotropic mouse hepatitis virus encephalomyelitis model, this study demonstrated an essential role of IFN-α/β receptor 1 (IFNAR1) specifically in neurons to control virus spread, regulate IFN-γ signaling, and prevent acute mortality. The results support the notion that effective neuronal IFNAR1 signaling compensates for their low basal expression of genes in the IFN-α/β pathway compared to glia. The data further highlight the importance of tightly regulated communication between the IFN-α/β and IFN-γ signaling pathways to optimize antiviral IFN-γ activity.

One proline deletion in the fusion peptide of neurotropic mouse hepatitis virus (MHV) restricts retrograde axonal transport and neurodegeneration

Mouse hepatitis virus (MHV; murine coronavirus) causes meningoencephalitis, myelitis, and optic neuritis followed by axonal loss and demyelination. This murine virus is used as a common model to study acute and chronic virus-induced demyelination in the central nervous system. Studies with recombinant MHV strains that differ in the gene encoding the spike protein have demonstrated that the spike has a role in MHV pathogenesis and retrograde axonal transport. Fusion peptides (FPs) in the spike protein play a key role in MHV pathogenesis. In a previous study of the effect of deleting a single proline residue in the FP of a demyelinating MHV strain, we found that two central, consecutive prolines are important for cell-cell fusion and pathogenesis. The dihedral fluctuation of the FP was shown to be repressed whenever two consecutive prolines were present, in contrast to the presence of a single proline in the chain. Using this proline-deleted MHV strain, here we investigated whether intracranial injection of this strain can induce optic neuritis by retrograde axonal transport from the brain to the retina through the optic nerve. We observed that the proline-deleted recombinant MHV strain is restricted to the optic nerve, is unable to translocate to the retina, and causes only minimal demyelination and no neuronal death. We conclude that an intact proline dyad in the FP of the recombinant demyelinating MHV strain plays a crucial role in translocation of the virus through axons and subsequent neurodegeneration.

Differential Regulation of DJ‐1 in Glial Cells upon MHV‐A59 ‐Induced Oxidative Stress

Astrocyte and microglial activation are two hallmarks associated with oxidative stress induction in neural cells. Generation of reactive oxygen species (ROS) is one of the molecular outcomes of neurotropic Mouse Hepatitis Virus A59 (MHV‐A59) infection in the Central Nervous System in an experimental animal model for human demyelinating disease Multiple Sclerosis. MHV‐A59 infection has been found to cause encephalitis, meningitis, and hepatitis in acute stage (Day 5–7 post‐infection) and progressive demyelination in chronic stage (Day 30 post‐infection). The virus has shown glial cell tropism leading to its activation, although the role of oxidative stress in this aspect is not well understood. Therefore, our work focuses on the regulation and function of important anti‐oxidative markers like DJ‐1, Nrf2, and HMOX‐1, upon viral infection in animal CNS tissue, isolated cells from tissues, primary culture and cell culture‐based studies. All three genes were found to be upregulated in vivo in mice brain at acute stage (Day 3 and 5) and chronic stage (Day 30) post‐infection. Similar upregulation was also observed in isolated microglia from mice brain, primary astrocytes, and microglia cultured from neonatal mice brain. In vitro studies with DBT astrocytoma cell line and N9 microglia cell line showed oxidative stress generation and upregulation in all three genes at initial time points of viral infection; however all the genes were found to be downregulated at higher time points. DBT cells exhibited decrease in cell viability and oxidative stress generation upon viral infection, and such changes had been observed to be ameliorated when DJ‐1 was stably transfected into DBT cells. Literature studies indicate that XBP1, an ER stress marker, upregulates DJ‐1 expression upon oxidative stress induction. In silico and in vitro studies showed that XBP1 was differentially activated in both DBT and N9 cells and was associated with DJ‐1 promoter upon viral infection in N9 cells. MHV‐A59 has shown tropism for glial cells resulting in their activation, and oxidative stress‐related cellular mechanisms involving these genes can contribute to the activation process of these glial cells.Support or Funding InformationThis study was supported by the Department of Biotechnology (DBT) and Indian Institute of Science Education and Research Kolkata (IISER Kolkata). Soumya Kundu was supported by University Grant Commission (UGC). Gisha Rose Anthony, Vineeth A. Raveendran, and Rahul Kumar were supported by DST‐INSPIRE fellowship under the banner of IISER Kolkata.

Pathogenic role of CD4+ T cells in the progression of neurotropic mouse hepatitis virus induced neuroinflammation

Abstract Neurotropic Mouse hepatitis virus (MHV) induced demyelination serves as a model to understand the mechanisms of demyelination and concomitant axonal loss in CNS, which mimics the pathological symptoms of the human neurological disease Multiple Sclerosis (MS). Inflammation triggered by MHV consists of mixed population of inflammatory cells, predominantly microglia/macrophages, which differs from autoimmune model of MS, where infiltrating T cells are the major contributors to the pathology. Though the acute phase of MHV inflammation is controlled by microglia/microglia, the role of T and B cells in viral induced axonal degeneration and demyelination is controversial. In order to explore the role of CD4+ T cells during transition from an early protective innate response to a highly regulated adaptive response following MHV infection, MHV-infected CD4−/− mice were compared with MHV-infected CD4+/+ (wildtype) mice at days 5/6, 10, 15 and 30 post infection. Viral titre estimation and nucleocapsid gene amplification confirm enhanced replication of the virions in the absence of functional CD4 + T cells in brain tissue. Viral anti-nucleocapsid antigen staining further shows the persistence of virus at day 30 p.i. in CD4−/− mice unlike CD4+/+ mice. Moreover, histopathological analysis of liver, brain and spinal cord showed an elevated susceptibility of CD4−/− mice to encephalitis and axonal degeneration with augmented progression of acute poliomyelitis, bulbar vacuolation, and dorsal root ganglionic inflammation rarely observed in CD4+/+ infected mice. In summary, current study suggests that CD4 T cell is critical for viral clearance, resolving microglial activation, and controlling chronic progressive axonal degeneration.

Demyelinating strain of mouse hepatitis virus infection bridging innate and adaptive immune response in the induction of demyelination

The presence of immunoglobulin oligoclonal bands in the cerebrospinal fluid of Multiple Sclerosis (MS) patients supports the hypothesis of an infectious etiology, although the antigenic targets remain elusive. Neurotropic mouse hepatitis virus (MHV) infection in mice provides a useful tool for studying mechanisms of demyelination in a virus-induced experimental model of MS. This study uses Affymetrix microarray analysis to compare differential spinal cord mRNA levels between mice infected with demyelinating and non-demyelinating strains of MHV to identify host immune genes expressed in this demyelinating disease model. The study reveals that during the acute stage of infection, both strains induce inflammatory innate immune response genes, whereas upregulation of several immunoglobulin genes during chronic stage infection is unique to infection with the demyelinating strain. Results suggest that the demyelinating strain induced an innate-immune response during acute infection that may promote switching of Ig isotype genes during chronic infection, potentially playing a role in antibody-mediated progressive demyelination even after viral clearance.

Role of inflammasome cytokines IL-1 and IL-18 during infection with neurotropic mouse hepatitis virus (INC8P.434)

Abstract The inflammasome is a cytosolic multiprotein complex that mediates secretion of pro-inflammatory cytokines IL-1 and IL-18. These cytokines have been found to play a role in restriction of a wide range of pathogens, and to contribute to the pathogenesis of many autoimmune conditions. IL-18 signaling has a known role in promoting production of interferon gamma from T and natural killer cells, while IL-1 polarizes Th17 responses and recruits neutrophils to sites of infection. However, work with influenza A and West Nile virus have shown that IL-1 signaling can also be necessary to promote strong anti-viral T cell responses via activation of dendritic cells. This effect was found to be specific to the site of infection. To further elucidate the roles of these cytokines we have infected mice lacking the IL-1 or IL-18 receptor with a dual neuro- and hepatotropic strain of mouse hepatitis virus (MHV). IL-18 receptor deficient mice succumb to normally non-lethal doses of MHV while IL-1 receptor deficient animals are somewhat protected from lethal infection, suggesting the cytokines play opposite roles to each other. The T cell response is similar in all genotypes, suggesting the cytokines act on innate populations of cells. We are currently investigating the behavior of the neutrophil and natural killer cell populations in inflammasome cytokine deficient mice. This work should elucidate the effect of inflammasome signaling on innate immune responses in differing sites of infections.

Microglia Play a Major Role in Direct Viral-Induced Demyelination

Microglia are the resident macrophage-like populations in the central nervous system (CNS). Microglia remain quiescent, unable to perform effector and antigen presentation (APC) functions until activated by injury or infection, and have been suggested to represent the first line of defence for the CNS. Previous studies demonstrated that microglia can be persistently infected by neurotropic mouse hepatitis virus (MHV) which causes meningoencephalitis, myelitis with subsequent axonal loss, and demyelination and serve as a virus-induced model of human neurological disease multiple sclerosis (MS). Current studies revealed that MHV infection is associated with the pronounced activation of microglia during acute inflammation, as evidenced by characteristic changes in cellular morphology and increased expression of microglia-specific proteins, Iba1 (ionized calcium-binding adaptor molecule 1), which is a macrophage/microglia-specific novel calcium-binding protein and involved in membrane ruffling and phagocytosis. During chronic inflammation (day 30 postinfection), microglia were still present within areas of demyelination. Experiments performed in ex vivo spinal cord slice culture and in vitro neonatal microglial culture confirmed direct microglial infection. Our results suggest that MHV can directly infect and activate microglia during acute inflammation, which in turn during chronic inflammation stage causes phagocytosis of myelin sheath leading to chronic inflammatory demyelination.

Surgical Transplantation of Mouse Neural Stem Cells into the Spinal Cords of Mice Infected with Neurotropic Mouse Hepatitis Virus

Mice infected with the neurotropic JHM strain of mouse hepatitis virus (MHV) develop pathological and clinical outcomes similar to patients with the demyelinating disease Multiple Sclerosis (MS). We have shown that transplantation of NSCs into the spinal cords of sick mice results in a significant improvement in both remyelination and in clinical outcome. Cell replacement therapies for the treatment of chronic neurologic diseases are now a reality and in vivo models are vital in understanding the interactions between the engrafted cells and host tissue microenvironment. This presentation provides an adapted method for transplanting cells into the spinal cord of JHMV-infected mice. In brief, we provide a procedure for i) preparation of NSCs prior to transplant, ii) pre-operative care of mice, iii) exposure of the spinal cord via laminectomy, iv) stereotactic injection of NSCs, and iv) post-operative care.

Surgical Transplantation of Mouse Neural Stem Cells into the Spinal Cords of Mice Infected with Neurotropic Mouse Hepatitis Virus

Mice infected with the neurotropic JHM strain of mouse hepatitis virus (MHV) develop pathological and clinical outcomes similar to patients with the demyelinating disease Multiple Sclerosis (MS). We have shown that transplantation of NSCs into the spinal cords of sick mice results in a significant improvement in both remyelination and in clinical outcome. Cell replacement therapies for the treatment of chronic neurologic diseases are now a reality and in vivo models are vital in understanding the interactions between the engrafted cells and host tissue microenvironment. This presentation provides an adapted method for transplanting cells into the spinal cord of JHMV-infected mice. In brief, we provide a procedure for i) preparation of NSCs prior to transplant, ii) pre-operative care of mice, iii) exposure of the spinal cord via laminectomy, iv) stereotactic injection of NSCs, and iv) post-operative care.

Pathogenesis of neurotropic murine coronavirus is multifactorial

Although coronavirus tropism is most often ascribed to receptor availability, increasing evidence suggests that for the neurotropic strains of the murine coronavirus mouse hepatitis virus (MHV), spike–receptor interactions cannot fully explain neurovirulence. The canonical MHV receptor CEACAM1a and its spike-binding site have been extensively characterized. However, CEACAM1a is poorly expressed in neurons, and the extremely neurotropic MHV strain JHM.SD infects ceacam1a−/− mice and spreads among ceacam1a−/− neurons. Two proposed alternative MHV receptors, CEACAM2 and PSG16, also fail to account for neuronal spread of JHM.SD in the absence of CEACAM1a. It has been reported that JHM.SD has an unusually labile spike protein, enabling it to perform receptor-independent spread (RIS), but it is not clear if the ability to perform RIS is fully responsible for the extremely neurovirulent phenotype. We propose that the extreme neurovirulence of JHM.SD is multifactorial and might include as yet unidentified neuron-specific spread mechanisms.

Viral nanoparticles associate with regions of inflammation and blood brain barrier disruption during CNS infection

Targeted treatment of inflammatory diseases of the central nervous system (CNS) remains problematic due to the complex pathogenesis of these disorders and difficulty in drug delivery. The plant virus, cowpea mosaic virus (CPMV), has recently been explored as a nanoparticle delivery system for therapeutics targeting a number of diseases including cancer and neurodegeneration. To understand the biodistribution of CPMV in the CNS, we examined CPMV uptake during infection of mice with neurotropic mouse hepatitis virus (MHV). CPMV localized mainly to the CNS endothelium in areas that contained an intact blood brain barrier. However, in inflammatory lesions containing macrophage/microglial cell infiltration and IgG, CPMV could be detected in the brain parenchyma. Furthermore, CPMV showed rapid internalization in an in vitro model of the BBB. These results suggest that CPMV particles could be used as a vehicle to deliver therapeutics to the damaged CNS during neurodegenerative and infectious diseases of the CNS.

The Role of IL-10 in Mice Infected with Neurotropic Murine Coronavirus (43.14)

Abstract Mice infected with the neurotropic coronavirus mouse hepatitis virus (MHV), attenuated strain J2.2 develop acute and chronic demyelinating disease with histopathological similarities to multiple sclerosis (MS). The process of demyelination is immune-mediated, as RAG1-/- or RAG2-/- (mice deficient in recombination activation enzyme), which lack B and T lymphocytes, do not develop demyelination upon infection with J2.2; however, if RAG1-/- mice are reconstituted with either MHV-immune CD4 or CD8 T cells, demyelination proceeded as in the immunocompetent mouse. While the pathogenic role of virus-specific CD4 and CD8 T cells and bystander CD8 T cells in this model has been demonstrated, less is known about the immune mechanisms that are controlling this anti-viral immune response. The cytokine IL-10 has been shown to be able to suppress an excessive immune response and protect the host against immune-mediated damage in EAE, an autoimmune model of multiple sclerosis. We investigated the role of IL-10 in demyelination using IL-10-/- mice. Additionally, we characterized the time, location, and antigen-specificity of IL-10 producing cells using IL-10gfp mice. Here, we demonstrate that IL-10 has a protective role in an animal model of MS and that the majority of IL-10 producing cells are MHV-specific T-cells located at the site of infection. Funded in part by grants from the NIH and National Multiple Sclerosis Society.

Memory CD4+T-Cell-Mediated Protection from Lethal Coronavirus Encephalomyelitis

The antiviral role of CD4(+) T cells in virus-induced pathologies of the central nervous system (CNS) has not been explored extensively. Control of neurotropic mouse hepatitis virus (JHMV) requires the collaboration of CD4(+) and CD8(+) T cells, with CD8(+) T cells providing direct perforin and gamma interferon (IFN-gamma)-mediated antiviral activity. To distinguish bystander from direct antiviral contributions of CD4(+) T cells in virus clearance and pathology, memory CD4(+) T cells purified from wild type (wt), perforin-deficient (PKO), and IFN-gamma-deficient (GKO) immune donors were transferred to immunodeficient SCID mice prior to CNS challenge. All three donor CD4(+) T-cell populations controlled CNS virus replication at 8 days postinfection, indicating IFN-gamma- and perforin-independent antiviral function. Recipients of GKO CD4(+) T cells succumbed more rapidly to fatal disease than untreated control infected mice. In contrast, wt and PKO donor CD4(+) T cells cleared infectious virus to undetectable levels and protected from fatal disease. Recipients of all CD4(+) T-cell populations exhibited demyelination. However, it was more severe in wt CD4(+) T-cell recipients. These data support a role of CD4(+) T cells in virus clearance and demyelination. Despite substantial IFN-gamma-independent antiviral activity, IFN-gamma was crucial in providing protection from death. IFN-gamma reduced neutrophil accumulation and directed macrophages to white matter but did not ameliorate myelin loss.

Pathogenesis of severe acute respiratory syndrome

Pathogenesis of severe acute respiratory syndrome

Coronaviruses: Molecular and Cellular Biology.

‘The knowledge contained in the book demonstrates how quickly and relatively efficiently the worldwide research community can meld together into a giant collaborative effort and concentrate its resources to rapidly begin to understand and solve problems, in this case a health problem.’ Coronaviruses cause respiratory and gastrointestinal diseases in various vertebrates, including minor respiratory diseases in humans. Because of that fact, focused research on these viruses was limited to a few dedicated, but talented, research groups. However, that changed in 2003, when a new coronavirus emerged, designated as severe acute respiratory syndrome corona-like virus (SARS-CoV), which caused a life-threatening respiratory disease characterized as an acute respiratory distress syndrome with a fatality rate of almost 10%. Suddenly, the world’s researchers devoted countless hours to attempting to identify, characterize and develop

CD4 T Cells Contribute to Virus Control and Pathology following Central Nervous System Infection with Neurotropic Mouse Hepatitis Virus

Replication of the neurotropic mouse hepatitis virus strain JHM (JHMV) is controlled primarily by CD8(+) T-cell effectors utilizing gamma interferon (IFN-gamma) and perforin-mediated cytotoxicity. CD4(+) T cells provide an auxiliary function(s) for CD8(+) T-cell survival; however, their direct contribution to control of virus replication and pathology is unclear. To examine a direct role of CD4(+) T cells in viral clearance and pathology, pathogenesis was compared in mice deficient in both perforin and IFN-gamma that were selectively reconstituted for these functions via transfer of virus-specific memory CD4(+) T cells. CD4(+) T cells from immunized wild-type, perforin-deficient, and IFN-gamma-deficient donors all initially reduced virus replication. However, prolonged viral control by IFN-gamma-competent donors suggested that IFN-gamma is important for sustained virus control. Local release of IFN-gamma was evident by up-regulation of class II molecules on microglia in recipients of IFN-gamma producing CD4(+) T cells. CD4(+) T-cell-mediated antiviral activity correlated with diminished clinical symptoms, pathology, and demyelination. Both wild-type donor CD90.1 and recipient CD90.2 CD4(+) T cells trafficked into the central nervous system (CNS) parenchyma and localized to infected white matter, correlating with decreased numbers of virus-infected oligodendrocytes in the CNS. These data support a direct, if limited, antiviral role for CD4(+) T cells early during acute JHMV encephalomyelitis. Although the antiviral effector mechanism is initially independent of IFN-gamma secretion, sustained control of CNS virus replication by CD4(+) T cells requires IFN-gamma.

Important Roles for Gamma Interferon and NKG2D in γδ T-Cell-Induced Demyelination in T-Cell Receptor β-Deficient Mice Infected with a Coronavirus

gammadelta T cells mediate demyelination in athymic (nude) mice infected with the neurotropic coronavirus mouse hepatitis virus strain JHM. Now, we show that these cells also mediate the same process in mice lacking alphabeta T cells (T-cell receptor beta-deficient [TCRbeta(-/-)] mice) and demyelination is gamma interferon (IFN-gamma) dependent. Most strikingly, our results also show a major role for NKG2D, expressed on gammadelta T cells, in the demyelinating process with in vivo blockade of NKG2D interactions resulting in a 60% reduction in demyelination. NKG2D may serve as a primary recognition receptor or as a costimulatory molecule. We show that NKG2D(+) gammadelta T cells in the JHM-infected central nervous system express the adaptor molecule DAP12 and an NKG2D isoform (NKG2D short), both required for NKG2D to serve as a primary receptor. These results are consistent with models in which gammadelta T cells mediate demyelination using the same effector cytokine, IFN-gamma, as CD8 T cells and do so without a requirement for signaling through the TCR.

Increased Epitope-Specific CD8+T Cells Prevent Murine Coronavirus Spread to the Spinal Cord and Subsequent Demyelination

CD8+ T cells are important for clearance of neurotropic mouse hepatitis virus (MHV) strain A59, although their possible role in A59-induced demyelination is not well understood. We developed an adoptive-transfer model to more clearly elucidate the role of virus-specific CD8+ T cells during the acute and chronic phases of infection with A59 that is described as follows. C57BL/6 mice were infected with a recombinant A59 virus expressing the gp33 epitope, an H-2Db-restricted CD8+ T-cell epitope encoded in the glycoprotein of lymphocytic choriomeningitis virus, as a fusion with the enhanced green fluorescent protein (RA59-gfp/gp33). P14 splenocytes (transgenic for a T-cell receptor specific for the gp33 epitope) were transferred at different times pre- and postinfection (p.i.). Adoptive transfer of P14 splenocytes 1 day prior to infection with RA59-gfp/gp33, but not control virus lacking the gp33 epitope, RA59-gfp, reduced weight loss and viral replication and spread in the brain and to the spinal cord. Furthermore, demyelination was significantly reduced compared to that in nonrecipients. However, when P14 cells were transferred on day 3 or 5 p.i., no difference in acute or chronic disease was observed compared to that in nonrecipients. Protection in mice receiving P14 splenocytes prior to infection correlated with a robust gp33-specific immune response that was not observed in mice receiving the later transfers. Thus, an early robust CD8+ T-cell response was necessary to reduce virus replication and spread, specifically to the spinal cord, which protected against demyelination in the chronic phase of the disease.

Astrocyte expression of a dominant-negative interferon-γ receptor

Interferon‐γ (IFN‐γ) is a major proinflammatory cytokine, and binding to its nearly ubiquitous receptor induces a wide variety of biological functions. To explore the role(s) of IFN‐γ signaling in astrocytes, transgenic mice (GFAP/IFN‐γR1ΔIC) expressing a dominant‐negative IFN‐γ receptor alpha chain under control of the astrocyte‐specific glial fibrillary acid protein (GFAP) promoter were generated. Transgenic mice developed normally, had normal astrocyte numbers and distribution, and exhibited no clinically overt phenotype. Transgene mRNA expression was detected only in the CNS, and the transgene‐encoded IFN‐γ receptor 1 colocalized with GFAP, which is consistent with astrocyte expression. Astrocytes from transgenic mice exhibited reduced IFN‐γ‐induced signaling as measured by major histocompatibility class II induction. Neither CNS inflammation nor perforin‐mediated clearance of a neurotropic mouse hepatitis virus from astrocytes was impaired following infection. Transgenic mice with impaired astrocyte responsiveness to IFN‐γ provide a model for studying the selective astrocyte‐dependent effects of this critical cytokine in CNS immunopathology. © 2005 Wiley‐Liss, Inc.