Exploring the VISTA of glial cells:astrocytes and microglia from development to disease

Multiple sclerosis (MS) is a chronic and often progressive disease of the central nervous system (CNS), which manifests at a young age. In MS, immune cell types including macrophages and lymphocytes enter the CNS and damage the myelin sheath, which acts as an insulation for axons and is crucial for rapid saltatory action potential propagation, and myelin damage also results in axonal loss. Immune checkpoints are receptors that provide a balance between enhancing and limiting the immune response to allow proper protective function without aberrant inflammation or autoimmune disease. Modulating immune checkpoint activity is a powerful tool to, for example, increase the immune response against cancer, or to reduce the immune response in autoimmune diseases. VISTA is an immune checkpoint that provides inhibitory signals to T cells leading to reduced immunity. Therefore, enhancing VISTA signaling in MS offers a novel treatment strategy to limit autoimmunity and reduce symptoms. In this thesis, the function of VISTA was investigated in CNS-resident cells, to explore the therapeutic potential of VISTA for MS. VISTA plays a role in the function of microglia, which are CNS-resident immune cells. Microglia have diverse functions in the CNS such as protection from intruders (e.g. bacteria and viruses), support of neurons, and tissue regeneration. In MS, microglia are involved in the anti-myelin immune response, but also in wound healing. VISTA regulates microglia uptake of myelin and microglia homeostasis. In conclusion, modulating VISTA signaling may present a novel strategy to treat MS, which likely affects CNS-resident cells

Exploring the VISTA of microglia:immune checkpoints in CNS inflammation

Negative checkpoint regulators (NCR) are intensely pursued as targets to modulate the immune response in cancer and autoimmunity. A large variety of NCR is expressed by central nervous system (CNS)-resident cell types and is associated with CNS homeostasis, interactions with peripheral immunity and CNS inflammation and disease. Immunotherapy blocking NCR affects the CNS as patients can develop neurological issues including encephalitis and multiple sclerosis (MS). How these treatments affect the CNS is incompletely understood, since expression and function of NCR in the CNS are only beginning to be unravelled. V-type immunoglobulin-like suppressor of T cell activation (VISTA) is an NCR that is expressed primarily in the haematopoietic system by myeloid and T cells. VISTA regulates T cell quiescence and activation and has a variety of functions in myeloid cells including efferocytosis, cytokine response and chemotaxis. In the CNS, VISTA is predominantly expressed by microglia and macrophages of the CNS. In this review, we summarize the role of NCR in the CNS during health and disease. We highlight expression of VISTA across cell types and CNS diseases and discuss the function of VISTA in microglia and during CNS ageing, inflammation and neurodegeneration. Understanding the role of VISTA and other NCR in the CNS is important considering the adverse effects of immunotherapy on the CNS, and in view of their therapeutic potential in CNS disease

Regionally diverse astrocyte subtypes and their heterogeneous response to EAE

Astrocytes fulfil many functions in the central nervous system (CNS), including contribution to the blood brain barrier, synapse formation, and trophic support. In addition, they can mount an inflammatory response and are heterogeneous in morphology and function. To extensively characterize astrocyte subtypes, we FACS-isolated and gene expression profiled distinct astrocyte subtypes from three central nervous system regions; forebrain, hindbrain and spinal cord. Astrocyte subpopulations were separated based on GLAST/SLC1A3 and ACSA-2/ATP1B2 cell surface expression. The local brain environment proved key in establishing different transcriptional programs in astrocyte subtypes. Transcriptional differences between subtypes were also apparent in experimental autoimmune encephalomyelitis (EAE) mice, where these astrocyte subtypes showed distinct responses. While gene expression signatures associated with blood-brain barrier maintenance were lost, signatures involved in neuroinflammation and neurotoxicity were increased in spinal cord astrocytes, especially during acute disease stages. In chronic stages of EAE, this reactive astrocyte signature was slightly decreased, while obtaining a more proliferative profile, which might be relevant for glia scar formation and tissue regeneration. Morphological heterogeneity of astrocytes previously indicated the presence of astrocyte subtypes, and here we show diversity based on transcriptome variation associated with brain regions and differential responsiveness to a neuroinflammatory insult (EAE)

Tomatidine reduces chikungunya virus progeny release by controlling viral protein expression

Author summaryChikungunya fever is a debilitating disease caused by the mosquito-borne Chikungunya virus. Over the past two decades the geographical spread of the virus and its mosquito vector has drastically increased thereby causing millions of infections. To date there is no antiviral drug and no vaccine available to treat/prevent Chikungunya virus infection. We recently showed that the natural steroidal alkaloid tomatidine has potent antiviral activity towards Chikungunya virus at submicromolar concentrations. In this study we dissected how tomatidine reduces the production of Chikungunya virus particles. We show that tomatidine lowers viral protein expression and we hypothesize that the effect of tomatidine on viral protein translation hampers the production of progeny viral RNA copies / number of infected cells thereby leading to a reduced production of secreted virus particles. Also, we show that Chikungunya virus does not readily become resistant to tomatidine. Collectively, we deciphered the mechanism by which tomatidine exerts antiviral activity to Chikungunya virus and our results strengthen the potential of tomatidine as an antiviral treatment strategy towards Chikungunya virus.Tomatidine, a natural steroidal alkaloid from unripe green tomatoes has been shown to exhibit many health benefits. We recently provided in vitro evidence that tomatidine reduces the infectivity of Dengue virus (DENV) and Chikungunya virus (CHIKV), two medically important arthropod-borne human infections for which no treatment options are available. We observed a potent antiviral effect with EC50 values of 0.82 mu M for DENV-2 and 1.3 mu M for CHIKV-LR. In this study, we investigated how tomatidine controls CHIKV infectivity. Using mass spectrometry, we identified that tomatidine induces the expression of p62, CD98, metallothionein and thioredoxin-related transmembrane protein 2 in Huh7 cells. The hits p62 and CD98 were validated, yet subsequent analysis revealed that they are not responsible for the observed antiviral effect. In parallel, we sought to identify at which step of the virus replication cycle tomatidine controls virus infectivity. A strong antiviral effect was seen when in vitro transcribed CHIKV RNA was transfected into Huh7 cells treated with tomatidine, thereby excluding a role for tomatidine during CHIKV cell entry. Subsequent determination of the number of intracellular viral RNA copies and viral protein expression levels during natural infection revealed that tomatidine reduces the RNA copy number and viral protein expression levels in infected cells. Once cells are infected, tomatidine is not able to interfere with active RNA replication yet it can reduce viral protein expression. Collectively, the results delineate that tomatidine controls viral protein expression to exert its antiviral activity. Lastly, sequential passaging of CHIKV in presence of tomatidine did not lead to viral resistance. Collectively, these results further emphasize the potential of tomatidine as an antiviral treatment towards CHIKV infection.Molecular basis of virus replication, viral pathogenesis and antiviral strategie

Exploring the VISTA of glial cells:astrocytes and microglia from development to disease

Multiple sclerosis (MS) is a chronic and often progressive disease of the central nervous system (CNS), which manifests at a young age. In MS, immune cell types including macrophages and lymphocytes enter the CNS and damage the myelin sheath, which acts as an insulation for axons and is crucial for rapid saltatory action potential propagation, and myelin damage also results in axonal loss. Immune checkpoints are receptors that provide a balance between enhancing and limiting the immune response to allow proper protective function without aberrant inflammation or autoimmune disease. Modulating immune checkpoint activity is a powerful tool to, for example, increase the immune response against cancer, or to reduce the immune response in autoimmune diseases. VISTA is an immune checkpoint that provides inhibitory signals to T cells leading to reduced immunity. Therefore, enhancing VISTA signaling in MS offers a novel treatment strategy to limit autoimmunity and reduce symptoms. In this thesis, the function of VISTA was investigated in CNS-resident cells, to explore the therapeutic potential of VISTA for MS. VISTA plays a role in the function of microglia, which are CNS-resident immune cells. Microglia have diverse functions in the CNS such as protection from intruders (e.g. bacteria and viruses), support of neurons, and tissue regeneration. In MS, microglia are involved in the anti-myelin immune response, but also in wound healing. VISTA regulates microglia uptake of myelin and microglia homeostasis. In conclusion, modulating VISTA signaling may present a novel strategy to treat MS, which likely affects CNS-resident cells

Exploring the VISTA of glial cells extended data

Extended data of PhD dissertation "Exploring the VISTA of glial cells

Exploring the VISTA of glial cells extended data

Extended data of PhD dissertation "Exploring the VISTA of glial cells

Systemic administration of beta-glucan induces immune training in microglia

Contains fulltext : 231625.pdf (publisher's version ) (Open Access