Rolle von Lymphotoxin Beta und Zelladhäsionsmolekül (CEACAM1) in Angeborene und adaptive Immunaktivierung

The induction of the adaptive and innate immune system is essential for controlling viral infections. Balance between the innate immune system and adaptive immune responses results in better antiviral immune activation. Not enough or too much activation can lead to impairment of immune responses. In this thesis, we have investigated the role of enforced viral replication and different mechanisms by which adequate innate and adaptive immune responses are generated. Recently, it was shown that Usp18 dependent viral replication in splenic CD169+ macrophages are important to induce a sufficient type I interferon response resulting in the subsequent activation of adaptive immune responses. Here, in one study, we have shown that the intracellular replication of viral particles is sufficient to generate virus specific CD8 T cell responses. On the other hand, extracellular distribution of viral particles along the splenic conduits was necessary for inducing systemic levels of type I interferon (IFN-I). We have shown that Usp18 is important for viral replication, but that B cell-derived lymphotoxin beta is necessary for the extracellular distribution of virus along the marginal zone in spleen. In a further study, we have demonstrated a role of CEACAM1 in B cell survival, proliferation and innate immune activation. B cells are important part of immune system because they are in close proximity of blood-borne antigen in splenic circulation, they secrete antibodies and maintain the lymphoid architecture. We have shown that cell intrinsic signalling of CEACAM1 is essential for survival of proliferating B cells. CEACAM1 is involved in B-cell receptor and signals through the Btk/Syk/NF-κB axis. Absence of CEACAM1 signalling leads to reduced number of B-cells in lymphoid organs especially marginal zone B cells and CD169+ macrophages. Absence of CEACAM1 leads to poor anti-viral antibody secretion after systemic infection with cytopathic virus resulting in early death of Ceacam1–/– mice. In another set of experiment, we investigated the role of virus specific antibodies on adaptive immune activation. With the use of chronic virus strain LCMV-Docile, we determined whether enforced viral replication could occur in presence of virus-specific antibodies or virus-specific CD8 T cells. We found that after systemic recall infection, virus-specific antibodies allow viral replication in splenic marginal zone but controlled in peripheral organs resulting in strong CD8 T cell priming whereas, virus-specific CD8 T cells blunted viral replication in spleen but failed to control the persistent chronic viral infection. In conclusion, using several mouse models of noncytopathic and cytopathic viral infection, we have elucidated the functional role of lymphotoxin beta, CEACAM1 and virus-specific antibodies in immune function particularly during enforced viral replication in the spleen.Die Aktivierung des adaptiven und angeborenen Immunsystems ist essentiell für die Kontrolle viraler Infektionen. Ausgeglichene Antworten des angeborenen und adaptiven Immunsystems führen zu einer verbesserten antiviraler Immunantwort. Zu geringe oder zu hohe Aktivierung führt zu einer verschlechterten Immunantwort. In dieser Arbeit wurde die Rolle der ‘’enforced virus replication’’ und verschiedene Mechanismen untersucht, welche eine adäquate angeborene und adaptive Immunaktivierung garantieren

Remote Data Acquisition and Visualization on an App

In order to allow power consumers to have a better understanding of their industrial electricity, it is necessary to provide them with real-time electricity information. Now a days we need to do meter reading, analysis manually. If any meter is goes down or fluctuating we need to do manual servicing and troubleshooting. It may cause meter loss, Time loss, and financial loss. The platform enables power consumers to have a better understanding of their own electricity consumption status and reasonable arrangements for use of electricity. Its deployment is very simple. For users, they do not need to change the household electricity structure. They can easily view the home electricity information and real-time monitor and control after installing power information visualization APP on their mobile phones or other mobile terminals. User interaction needs to collect power information of all equipment, and provide users with real-time electricity price and electricity information. The basic purpose of system is M-Measure, A-Analysis, R-Record, and CControl. No human intervention for data processing analysis. This is use for Power quality analysis, Identification and prevention of downtime of assets, Alarms and triggers are used for preventive control to avoid loss. Reduce the losses by monitoring and controlling the power quality. Accurate analytics depending on the load connected to the system. It supports most of all make sensors and hardware meters to work with the system. Periodical analysis of the organization for the complete energy health check-up

Virus-induced type I interferon deteriorates control of systemic pseudomonas aeruginosa infection

BACKGROUND: Type I interferon (IFN-I) predisposes to bacterial superinfections, an important problem during viral infection or treatment with interferon-alpha (IFN-α). IFN-I-induced neutropenia is one reason for the impaired bacterial control; however there is evidence that more frequent bacterial infections during IFN-α-treatment occur independently of neutropenia. METHODS: We analyzed in a mouse model, whether Pseudomonas aeruginosa control is influenced by co-infection with the lymphocytic choriomeningitis virus (LCMV). Bacterial titers, numbers of neutrophils and the gene-expression of liver-lysozyme-2 were determined during a 24 hours systemic infection with P. aeruginosa in wild-type and Ifnar(-/-) mice under the influence of LCMV or poly(I:C). RESULTS: Virus-induced IFN-I impaired the control of Pseudomonas aeruginosa. This was associated with neutropenia and loss of lysozyme-2-expression in the liver, which had captured P. aeruginosa. A lower release of IFN-I by poly(I:C)-injection also impaired the bacterial control in the liver and reduced the expression of liver-lysozyme-2. Low concentration of IFN-I after infection with a virulent strain of P. aeruginosa alone impaired the bacterial control and reduced lysozyme-2-expression in the liver as well. CONCLUSION: We found that during systemic infection with P. aeruginosa Kupffer cells quickly controlled the bacteria in cooperation with neutrophils. Upon LCMV-infection this cooperation was disturbed

Role of NK Cells in Cancer and Immunotherapy

Increasing knowledge of cancer immunology has led to the design of therapies using immune cells directly or manipulating their activity, collectively termed immunotherapy. In the field of immuno-oncology, research on adaptive immune T cells has led to the development of CAR-T cells. Innate immune cells such as NK cells can also eliminate oncogenically transformed cells and regulate cells of the immune system. Considering NK cells as a live drug, numerous methods for the isolation and activation of NK cells have been shown to be clinically and therapeutically relevant. In such processes, various cytokines and antibodies present a source of stimulation of NK cells and enhance the efficacy of such treatments. The ex vivo expansion and activation of NK cells, along with genetic modification with CAR, enhance their antitumor activity. Recent preclinical studies have shown an antitumor effect through extracellular vesicles (EVs) derived from NK cells. Work with autologous NK cells has provided insights for clinical applications. In this review, we outline the recent advances of NK-cell-based immunotherapies, summarizing CAR-NK cells, BiKEs, and TriKEs as treatment options against cancer. This review also discusses the challenges of NK cell immunotherapy

Expression of JAK3 Sensitive Na+^+ Coupled Glucose Carrier SGLT1 in Activated Cytotoxic T Lymphocytes

Background: Similar to tumor cells, activated T-lymphocytes generate ATP mainly by glycolytic degradation of glucose. Lymphocyte glucose uptake involves non-concentrative glucose carriers of the GLUT family. In contrast to GLUT isoforms, Na+-coupled glucose-carrier SGLT1 accumulates glucose against glucose gradients and is effective at low extracellular glucose concentrations. The present study explored expression and regulation of SGLT1 in activated murine splenic cytotoxic T cells (CTLs) and human Jurkat T cells. Methods: FACS analysis, immunofluorescence, confocal microscopy, chemiluminescence and Western blotting were employed to estimate SGLT1 expression, function and regulation in lymphocytes, as well as dual electrode voltage clamp in SGLT1 ± JAK3 expressing Xenopus oocytes to quantify the effect of janus kinase3 (JAK3) on SGLT1 function. Results: SGLT1 is expressed in murine CTLs and also in human Jurkat T cells. 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose uptake was significantly decreased by SGLT1-blocker phloridzin (0.2 mM) and by pharmacological inhibition of JAK3 with WHI-P131 (156 µM), WHI-P154 (11.2 µM) and JAK3 inhibitor VI (0.5 µM). Electrogenic glucose transport (Iglucose) in Xenopus oocytes expressing human SGLT1 was increased by additional expression of human wild type JAK3, active A568VJAK3 but not inactive K851AJAK3. Coexpression of JAK3 enhanced the maximal transport rate without significantly modifying affinity of the carrier. Iglucose in SGLT1+JAK3 expressing oocytes was significantly decreased by WHI-P154 (11.2 µM). JAK3 increased the SGLT1 protein abundance in the cell membrane. Inhibition of carrier insertion by brefeldin A (5 µM) in SGLT1+JAK3 expressing oocytes resulted in a decline of Iglucose, which was similar in presence and absence of JAK3. Conclusions: SGLT1 is expressed in murine cytotoxic T cells and human Jurkat T cells and significantly contributes to glucose uptake in those cells post activation. JAK3 up-regulates SGLT1 activity by increasing the carrier protein abundance in the cell membrane, an effect enforcing cellular glucose uptake into activated lymphocytes and thus contributing to the immune response

Integrin Alpha E (CD103) Limits Virus-Induced IFN-I Production in Conventional Dendritic Cells.

Early and strong production of IFN-I by dendritic cells is important to control vesicular stomatitis virus (VSV), however mechanisms which explain this cell-type specific innate immune activation remain to be defined. Here, using a genome wide association study (GWAS), we identified Integrin alpha-E (Itgae, CD103) as a new regulator of antiviral IFN-I production in a mouse model of vesicular stomatitis virus (VSV) infection. CD103 was specifically expressed by splenic conventional dendritic cells (cDCs) and limited IFN-I production in these cells during VSV infection. Mechanistically, CD103 suppressed AKT phosphorylation and mTOR activation in DCs. Deficiency in CD103 accelerated early IFN-I in cDCs and prevented death in VSV infected animals. In conclusion, CD103 participates in regulation of cDC specific IFN-I induction and thereby influences immune activation after VSV infection

SGLT1 Deficiency Turns Listeria Infection into a Lethal Disease in Mice

Background: Cellular glucose uptake may involve either non-concentrative glucose carriers of the GLUT family or Na+-coupled glucose-carrier SGLT1, which accumulates glucose against glucose gradients and may thus accomplish cellular glucose uptake even at dramatically decreased extracellular glucose concentrations. SGLT1 is not only expressed in epithelia but as well in tumour cells and immune cells. Immune cell functions strongly depend on their metabolism, therefore we hypothesized that deficiency of SGLT1 modulates the defence against bacterial infection. To test this hypothesis, we infected wild type mice and gene targeted mice lacking functional SGLT1 with Listeria monocytogenes. Methods: SGLT1 deficient mice and wild type littermates were infected with 1x104 CFU Listeria monocytogenes intravenously. Bacterial titers were determined by colony forming assay, SGLT1, TNF-α, IL-6 and IL-12a transcript levels were determined by qRT-PCR, as well as SGLT1 protein abundance and localization by immunohistochemistry. Results: Genetic knockout of SGLT1 (Slc5a1–/– mice) significantly compromised bacterial clearance following Listeria monocytogenes infection with significantly enhanced bacterial load in liver, spleen, kidney and lung, and significantly augmented hepatic expression of TNF-α and IL-12a. While all wild type mice survived, all SGLT1 deficient mice died from the infection. Conclusions: SGLT1 is required for bacterial clearance and host survival following murine Listeria infection

SGLT1 Deficiency Turns Listeria Infection into a Lethal Disease in Mice

Background: Cellular glucose uptake may involve either non-concentrative glucose carriers of the GLUT family or Na+-coupled glucose-carrier SGLT1, which accumulates glucose against glucose gradients and may thus accomplish cellular glucose uptake even at dramatically decreased extracellular glucose concentrations. SGLT1 is not only expressed in epithelia but as well in tumour cells and immune cells. Immune cell functions strongly depend on their metabolism, therefore we hypothesized that deficiency of SGLT1 modulates the defence against bacterial infection. To test this hypothesis, we infected wild type mice and gene targeted mice lacking functional SGLT1 with Listeria monocytogenes. Methods: SGLT1 deficient mice and wild type littermates were infected with 1x104 CFU Listeria monocytogenes intravenously. Bacterial titers were determined by colony forming assay, SGLT1, TNF-α, IL-6 and IL-12a transcript levels were determined by qRT-PCR, as well as SGLT1 protein abundance and localization by immunohistochemistry. Results: Genetic knockout of SGLT1 (Slc5a1–/– mice) significantly compromised bacterial clearance following Listeria monocytogenes infection with significantly enhanced bacterial load in liver, spleen, kidney and lung, and significantly augmented hepatic expression of TNF-α and IL-12a. While all wild type mice survived, all SGLT1 deficient mice died from the infection. Conclusions: SGLT1 is required for bacterial clearance and host survival following murine Listeria infection

Two separate mechanisms of enforced viral replication balance innate and adaptive immune activation

The induction of innate and adaptive immunity is essential for controlling viral infections. Limited or overwhelming innate immunity can negatively impair the adaptive immune response. Therefore, balancing innate immunity separately from activating the adaptive immune response would result in a better antiviral immune response. Recently, we demonstrated that Usp18-dependent replication of virus in secondary lymphatic organs contributes to activation of the innate and adaptive immune responses. Whether specific mechanisms can balance innate and adaptive immunity separately remains unknown. In this study, using lymphocytic choriomeningitis virus (LCMV) and replication-deficient single-cycle LCMV vectors, we found that viral replication of the initial inoculum is essential for activating virus-specific CD8(+) T cells. In contrast, extracellular distribution of virus along the splenic conduits is necessary for inducing systemic levels of type I interferon (IFN-I). Although enforced virus replication is driven primarily by Usp18, B cell-derived lymphotoxin beta contributes to the extracellular distribution of virus along the splenic conduits. Therefore, lymphotoxin beta regulates IFN-I induction independently of CD8(+) T-cell activity. We found that two separate mechanisms act together in the spleen to guarantee amplification of virus during infection, thereby balancing the activation of the innate and adaptive immune system

TLR7 controls VSV replication in CD169(+) SCS macrophages and associated viral neuroinvasion

Vesicular stomatitis virus (VSV) is an insect-transmitted rhabdovirus that is neurovirulent in mice. Upon peripheral VSV infection, CD169+ subcapsular sinus (SCS) macrophages capture VSV in the lymph, support viral replication, and prevent CNS neuroinvasion. To date, the precise mechanisms controlling VSV infection in SCS macrophages remain incompletely understood. Here, we show that Toll-like receptor-7 (TLR7), the main sensing receptor for VSV, is central in controlling lymph-borne VSV infection. Following VSV skin infection, TLR7−/− mice display significantly less VSV titers in the draining lymph nodes (dLN) and viral replication is attenuated in SCS macrophages. In contrast to effects of TLR7 in impeding VSV replication in the dLN, TLR7−/− mice present elevated viral load in the brain and spinal cord highlighting their susceptibility to VSV neuroinvasion. By generating novel TLR7 floxed mice, we interrogate the impact of cell-specific TLR7 function in anti-viral immunity after VSV skin infection. Our data suggests that TLR7 signaling in SCS macrophages supports VSV replication in these cells, increasing LN infection and may account for the delayed onset of VSV-induced neurovirulence observed in TLR7−/− mice. Overall, we identify TLR7 as a novel and essential host factor that critically controls anti-viral immunity to VSV. Furthermore, the novel mouse model generated in our study will be of valuable importance to shed light on cell-intrinsic TLR7 biology in future studies