bluuebunny/crossref_metadata_2025_split huggingface dataset:
A role for Mitochondrial Antiviral Signaling molecule (MAVS) in the TCR pathway.
Authors: Thomas Sunil , Sekine Aimee , Gale Michael 
Published: January 2023

Abstract
Mitochondrial Antiviral Signaling molecule (MAVS) is an adaptor that functions downstream of the viral RNA sensors RIG-I and MDA5 to translate viral RNA sensing into effector responses such as the production of Type I IFN and inflammatory cytokines that result in the anti viral state that functions to limit viral spread. However a precise role for MAVS in CD8 T cells has not been delineated.
Using an in vivo adoptive transfer system with Lymphocytic choriomeningitis virus (LCMV) and transgenic LCMV- GP33 specific T cells (P14) and OVA-specific T cells (OT1) in vitro, we show that there is a cell-intrinsic role for MAVS in transducing signals proximal to the T cell receptor in both in vivo and in vitro settings. MAVS-dependent defects in CD8 T cell effector functions were identified in cells responding to in vivo LCMV infection in mice. At the peak of the adaptive response to LCMV, MAVS-deficient CD8 T cells were smaller in size, had deficient intracellular calcium, decreased mitochondrial membrane potential, decreased NFATc1 as well as defective production of the effector cytokines IFN γ, TNF α and IL-2. We found that MAVS is involved in the TCR signaling pathway by regulating the levels of phosphorylated zeta chain of protein kinase 70 (pZAP70), an enzyme that functions in the initial step of TCR-mediated signal transduction. The levels of pZAP70 were lower in the absence of MAVS in vivo in P14 CD8 T cells as well as in in vitro stimulated OT1 CD8 T cells. The degree of defect was dynamic and varied as a function of time after TCR ligation. The precise mechanisms by which MAVS regulates pZAP70 levels will be presented.
Failure to control measles virus spread in the neonatal CNS despite immune cell infiltration and cytokine production.
Authors: Ganesan Priya , Bohn Larissa , O’Donnell Lauren 
Published: January 2023

Abstract
Viral infections in the central nervous system (CNS) are associated with devastating neurological consequences in newborns. This may be due, in part, to deficiencies in the immature innate and adaptive immune response. To study viral CNS infections in neonates, we use a transgenic mouse model of neuronally-restricted measles virus (MV) infection (NSE-CD46 mice), where the human isoform of CD46, a MV receptor, is expressed under the control of the neuron-specific enolase promoter. Adult CD46+ mice survive infection and clear MV in an interferon gamma (IFNg)- and T cell-dependent manner. In contrast, neonatal CD46+ mice succumb to the infection despite T cell infiltration into the brain parenchyma. Neonatal mice lacking IFNg succumb more rapidly than wildtype pups despite higher T cell infiltration, equivalent natural killer cell infiltration, and microglial activation. Quantitative RT-PCR analysis demonstrated upregulation of pro-inflammatory cytokines and chemokines such as IFNg, IL-1β, and CXCL10, as well as the anti-inflammatory interleukin 1 receptor antagonist in infected adults and neonates. IL-12β, IL-23, CCL2, and TNF were significantly upregulated in infected neonates only. These results suggest the neonates are capable of cytokine production with a Th1-like profile within the CNS, but that the cytokine milieu is ineffective at controlling MV spread. Current experiments aim to inhibit inflammatory mediators such as CXCL10 and CCL2 that are expressed in an age-dependent manner during infection to determine if there is an improvement in neuropathology and survival of neonates.
Expression and regulatory effects of Schlafen genes in flavivirus infection
Authors: Kumar Mukesh , Hughes Laura , Nerurkar Vivek R 
Published: January 2023

Abstract
The continuing spread of flaviviruses, combined with the lack of specific therapeutics and vaccines, imparts a pressing need to identify the viral and host factors that control replication and outcome of infection. The Schlafen (SLFN) family includes several mouse and human genes that have been investigated for their role in cell proliferation, regulation of immune response, and control of virus replication. Role of SLFN family genes in flavivirus infections is yet to be determined. Herein, we demonstrate that infection with lethal West Nile virus (WNV) NY99 strain induces the expression of SLFN family genes; SLFN2, SLFN3, SLFN4, SLFN5, SLFN8, and SLFN9, in mice brain, and mouse primary cells including mouse embryonic fibroblasts (MEFs), macrophages, astrocytes, neurons and microglia. Among SLFN family genes, highest induction was observed in the expression of SLFN4. Interestingly, infection with non-lethal WNV Eg101 strain does not induce the expression of SLFN family genes in mice brain. Moreover, WNV NY99 infection of primary human cells including neurons, astrocytes, brain endothelial cells induced dramatic up regulation of SLFN12, SLFN12L (human orthologs of SLFN4), and SLFN14. Similarly, infection with dengue virus-1 (DENV-1), DENV-2, DENV-3, and DENV-4 induced the expression of SLFN12, SLFN12L and SLFN14 in Vero cells and human monocytes. Furthermore, using siRNA, we demonstrate that knockdown of SLFN4 results in significant decrease of WNV NY99 titers in MEFs. Additionally, mRNA and protein expression of WNV-induced interferons, chemokines, and cytokines were significantly decreased in SLFN4-knockdown MEFs. Collectively our data for the first time indicate the novel role of SLFN genes in flavivirus pathogenesis.
Interactions between the Ranavirus Frog virus 3 and mammalian macrophage cell lines
Authors: Torres-Luquis Odalys Joan , Edholm Eva-Stina , De Jesús-Andino Francisco , Elliott Michael Rusty , Robert Jacques 
Published: January 2023

Abstract
Infections caused by Ranaviruses (RVs, Irridoviridae) are increasing at alarming rates in prevalence and the wide range of host species infected, (from amphibians to fishes and reptiles), which suggests that RVs possess potent immune evasion strategies. Growing evidence indicated that RVs host infection strategies and disseminations are critically linked to the host immune response, specifically macrophage-lineage subsets. Indeed, we have shown that in the amphibian Xenopus laevis the RV Frog virus 3 (FV3) can persist in a quiescent state in macrophages of otherwise asymptomatic hosts and can be reactivated following inflammation-induced stimulation. To further explore the cellular and molecular mechanisms involved in these processes we have developed an in vitro culture system using FV3 in combination with five murine macrophage cell lines that were adapted to 30°C culture condition necessary for FV3 replication. FV3 infectivity was determined using a knock-in recombinant FV3 expressing a GFP reporter under the control of the 18K immediate-early FV3 promoter. FV3 infected all murine cell lines. However, viral replication varied depending on the cell type: it was similar to that detected in control BHK cells with RAW (peritoneal) cells, comparably modest with J774 (peritoneal) and BV-2 (microglial) cells, whereas no FV3 replication was detected with TM4 (Sertoli) and P338D1 (monocytic) cells. This lack of FV3 replication in TM4 and P338D1 despite successful viral entry and expression of FV3 immediate-early genes in TM4 is similar to what is observed with X. laevis macrophages and suggests that FV3 triggers similar cellular and molecular mechanisms in mammalian as in X. laevis macrophages resulting in viral quiescence.
Resveratrol (RES) and its antagonist, sirtinol suppress human cytomegalovirus (hCMV) replication and hCMV induced transcription of proinflammatory cytokines and chemokines in THP-1 derived macrophages
Authors: Li Huifen , Ning Xiaoxuan , Wang Jintang , Ouyang Min , Mao Genxiang , Leng Sean X 
Published: January 2023

Abstract

Background
Mounting evidence suggests significant contribution of chronic hCMV infection to T-cell immunosenenscence and chronic inflammation in old adults. THP-1, a human monocytic cell line and its derived macrophages serve as a useful cell culture model to study hCMV infection and its biology. Novel anti-CMV agents are needed as current anti-CMV treatment is limited in efficacy with significant side effects. We and others observed that Resveratrol (RES) and sirtinol, suppressed CMV replication in human fibroblasts. Here we report their inhibition on hCMV replication and hCMV induced expression of IL-6, IP-10, MCP-1 and TNF-a in THP-1 derived macrophages.

Methods
THP-1 cells were differentiated into macrophages after incubation with PMA (50 ng/ml) in RPMI 1640 with 10%FBS for 24 hrs. RES or sirtinol was added 2 h followed by hCMV (Towne strain) inoculation at multiplicity of infection of MOI 2 for 24 hrs. Cells were then cultured in hCMV-free medium for indicated times. qPCR and RT-PCR were performed to evaluate hCMV replication and transcription of IL-6, IP-10, MCP-1 and TNF-a, respectively.

Results
Both RES and sirtinol suppressed hCMV replication. They also suppressed hCMV induced transcription of proinflammatory mediators except for TNF-a.

Conclusions and discussion
These findings indicate potent inhibition of hCMV infection and hCMV induced transcription of proinflammatory mediators by RES and sirtinol in THP-1 derived macrophages. While being non-permissive, monocytes are important reservoir for chronic or latent CMV infection. Further investigation into the role and mechanisms of RES, sirtinol and other agents in control of chronic CMV infection and its adverse impact is indicated.
Markers of Chronic Immune Activation in HIV Patients Receiving Antiretroviral Therapy
Authors: Albino Elinette , Godoy Lenin , Hill Martin 
Published: January 2023

Abstract
Antiretroviral therapy (ART) prolongs the lifespan of HIV-infected individuals but does not reduce chronic immune activation and inflammation. While ART typically decreases both immune activation and inflammation markers, recent studies showed these markers remain elevated in many HIV-infected patients. Over time, this heightened inflammation state may increase the rate of cardiovascular disease in HIV-infected individuals. In this study we characterize the inflammatory monocyte subsets in HIV positive subjects taking HAART and correlate this information with innate and inflammatory cytokines levels in plasma. Blood samples were taken from 50 subjects including: HIV positive subjects under ART, untreated HIV positive subjects healthy subjects taking HAART and healthy controls. Nine cytokines were measured in plasma using a Magnetic Luminex technology. In addition, fresh monocytes were separated from peripheral blood mononuclear cells (PBMCs) by CD14/CD16 monoclonal antibodies. Levels inflammatory cytokines were associated with health status parameters and self reported lifestyles. HIV-1 infected subjects had increased levels of both CD14/CD16 cells and inflammatory cytokines compare to the rest of the subjects enrolled in the study. A plasma inflammatory biomarker signature consisting of CD14, IL-6, IL-15, CD163 and IL-12 p70 corresponded with immune activation after prolonged exposition to ART. These findings suggest that increased inflammatory cytokines levels could be a consequence of antiretroviral use, yet the specific role of these inflammatory markers in disease state remains to be determined.
RIPK3 restricts West Nile virus neuropathogenesis independently of programmed necrosis.
Authors: Daniels Brian , Loo Yueh-Ming , Gale Michael , Oberst Andrew 
Published: January 2023

Abstract
Programmed necrosis coordinated by receptor-interacting kinase-3 (RIPK3) and mixed lineage kinase domain-like protein (MLKL) is known to contribute to host immune responses to viral infections. However, the role of these proteins in host immunity during flavivirus infection remains poorly understood. Using a mouse model of West Nile virus encephalitis, we show that RIPK3 is required for the restriction of West Nile virus pathogenesis, independently of its role in programmed necrosis. While Ripk3−/− mice exhibited accelerated and enhanced mortality compared to WT controls, survival and clinical scores in mice lacking the executioner protein MLKL were unaffected. Moreover, cell death following WNV infection in primary cultures of myeloid and neuronal cells was unchanged in both Ripk3−/− and Mlkl−/− cultures. The enhanced susceptibility of Ripk3−/− mice arose, instead, from a failure to control infection within the central nervous system (CNS). While peripheral adaptive immune responses to WNV remained intact, Ripk3−/− mice exhibited decreased recruitment of inflammatory myeloid cells and lymphocytes to the CNS, despite higher CNS viral loads. RIP3 suppressed CNS viral burden via both CNS-intrinsic and –extrinsic mechanisms, as viral titers in the brain and spinal cord were higher in Ripk3−/− mice compared to controls following both subcutaneous and intracranial inoculation with WNV. Together, these data identify new roles for RIPK3 in the restriction of viral pathogenesis, and implicate RIPK3 as a key regulator of antiviral immunity within the CNS.
ZMPSTE24 is a novel intrinsic immune protein that restricts a broad-spectrum of viruses
Authors: Li Shitao , Wang Lingyan , Fu Bishi , Dorf Martin E 
Published: January 2023

Abstract
Virus entry begins with attachment to cell-surface receptors and ends with delivery of the viral genome into the cytoplasm. Many enveloped viruses are ferried deep into the cytoplasm through endocytic vesicles and utilize the low pH endosomal environment to activate the fusion process between viral and cellular membranes. Higher vertebrates evolved interferon-induced transmembrane proteins (IFITM) to impede endocytic penetration by a broad spectrum of RNA viruses. However, the molecular mechanism by which IFITM proteins hinder viral entry is not well elucidated. To address the antiviral mechanism of IFITM, we hypothesize that type I interferon (IFN) induces recruitment of co-factors to the IFITM3 protein complex. Proteomic analysis of IFITM3-FLAG protein complex in HEK293 cells revealed that IFN induced IFITM3 to recruit ZMPSTE24, the 7-pass transmembrane zinc metalloprotease whose function in cytoplasmic organelles is not defined. Contrary to IFITM, ZMPSTE24 mRNA and protein expression are not regulated by IFN. But consistent with IFITM, ZMPSTE24 restricts both RNA and DNA viruses using endocytic entry, including influenza A, vesicular stomatitis, sindbis, Ebola, cowpox and vaccinia viruses. Antiviral activity requires ZMPSTE24 endosomal localization and is independent of its protease activity. Furthermore, ZMPSTE24 is recruited to endosomes by IFITM and block viral endocytic entry. These findings identify a new role for ZMPSTE24 as a broad-spectrum intrinsic antiviral protein and the downstream antiviral effector of IFITM.
MHC class II antigen-processing chaperone H2-O shapes CD4 T cell receptor repertoire
Authors: Song Nianbin , Poluektov Yuri , Welsh Robin , Sadegh-Nasseri Scheherazade 
Published: January 2023

Abstract
HLA-DO (DO) is a MHC class II-encoded, nonpolymorphic heterodimeric protein in humans (known as H2-O in mice). It is expressed only in thymic medulla, B lymphocytes and some subsets of dendritic cells. Although DO is found to form a stable complex with HLA—DM (DM), its physical function remains unknown. Some studies have shown that it inhibits the activity of DM to edit peptide presentation on MHC II molecules while others reported that the DO can up- or downmodulate presentation of certain peptides in association with DM on HLA-DR4 molecules. To solve this controversy, our lab recently proposed a model suggesting that the DO facilitates the binding of DM-resistant peptides and inhibits the binding of DM-sensitive peptides to the HLA-DR1 molecules in vitro. Based on this model, we expected to see lower density of pMHC expression on mTEC cells in thymus of H2-O-KO mice, which will lead to faulty negative selection and different CD4 TCR repertoires as compared to H2-O-WT mice. By developing a “Prime-Restim” strategy based on the principle of mixed lymphocyte reaction, we observed the proliferation of primed H2-O-WT CD4 T cells in responding to repeated in vitro stimulations by antigen presenting cells from H2-O-KO mice and vice versa. The results demonstrate that in the absence of H2-O, the peptide repertoire of MHC II in the H2-O knockout mice is different from the peptide repertoire of MHC II in the H2-O-WT mice. It establishes a role for DO in regulation of epitope selection for presentation by MHC class II molecules in vivo. These findings support the idea that the expression of DO in thymic medulla might lead to better thymic deletion of self-reactive CD4 T cells.
MARCH1 ubiquitin ligase rescues thymic dendritic cells from MHCII proteotoxicity and supports regulatory T cell development and repertoire expansion
Authors: Oh Jaehak , Perry Justin , Pua Heather , Hsieh Chyi-song , Shin Jeoung-Sook 
Published: January 2023

Abstract
Membrane-Associated RING-CH1 (MARCH1) is an ubiquitin ligase preferentially expressed in antigen-presenting cells and mediating ubiquitination and degradation of MHCII and CD86. MARCH1 deficiency results in a significant reduction in the number of thymic regulatory T (Treg) cells in mice, implicating an important role of this molecule in Treg cell development. However, neither the mechanism by which MARCH1 supports Treg cell development nor its impact on Treg cell function has been clearly defined. Here we report that the role of MARCH1 in Treg cell development critically depends on its expression in dendritic cells and its activity to ubiquitinate MHCII. Importantly, not the MHCII ubiquitination per se but its consequences to MHCII proteostasis is essential for dendritic cells to organize surface proteins appropriately, make a stable conjugate with thymocytes, and provide them with sufficient signal for Treg cell differentiation. When MARCH1-mediated MHCII proteostasis in dendritic cells fails, Treg cells develop to a markedly reduced number and a substantially restricted TCR repertoire in the thymus, resulting in the function of these cells to suppress graft-versus-host-disease severely impaired. Thus, MARCH1-mediated MHCII proteostasis plays an essential role for thymic dendritic cells to select regulatory T cells in abundance and diversity required for proper immune suppression.
The future of the immunopeptidome in health and disease: a comprehensive analysis of naturally processed ligand data
Authors: Vaughan Kerrie , Caron Etienne , Peters Bjoern , Sette Alessandro 
Published: January 2023

Abstract
The promise of big data (‘OMICS’) in fundamental and translational science is clear. In this respect, high-throughput analysis of HLA class I and II associated immunopeptidomes by mass spectrometry is an important tool to establish T cell response repertoires in the context of disease. The last decade has seen tremendous growth in the breadth and number of these studies. The time is therefore ideal for initial assessment of these data as a whole. For this, we made use of all naturally eluted peptide data captured to date within the IEDB, representing more than 100,000 class I and II peptides. The goal of this analysis was to compare and contrast the overall nature of naturally eluted peptides (NP) from several standpoints, including molecular and biological function, sub-cellular location and protein class. Further, we investigated the extent to which the NP data overlap with existing T cell epitope data, MHC binding data, as well as output from class I and II prediction algorithms. The comprehensive collection of NP data will allow us to examine and bench mark the correlation with independently determined immunological response and MHC binding, and might also be useful to develop novel methods for the identification of optimal target antigens and for epitope prediction. These data support the potential value of immunopeptidomics in understanding T cell immunity.
Systemic recognition of insulin by autoimmune T and B cells leads to production of antibodies targeting the islet of Langerhans
Authors: Wan Xiaoxiao , Thomas James W , Unanue Emil R 
Published: January 2023

Abstract
The presence of class-switched insulin autoantibodies (IAAs) is a consistent feature of patients with type 1 diabetes and non-obese diabetic mice. The mechanisms underlying their production and pathogenic role are poorly understood. IAA production is unexpected because insulin-reactive B cells are maintained in a tolerant state. Furthermore, the source of T cell help is limited because T cells recognizing insulin are deleted in the thymus, and those that escape thymic selection recognize insulin peptides but not native insulin. However, increased titers of IAAs to conformational insulin were found in a T cell receptor transgenic strain, 8F10, whose T cell reactivity is restricted to insulin peptides but not insulin protein. Furthermore, when an anti-insulin B cell receptor heavy chain transgene was introduced into the 8F10 mice, a marked increase in IAAs specific for native insulin was observed. Dynamic T-B cell interactions and spontaneous germinal center formation were observed in multiple lymphoid organs indicating widespread insulin presentation. But more pointedly we found that the germinal center B cells were activated and in contrast to naïve B cells, presented a broader repertoire of insulin epitopes. Finally, for the first time we show that IAAs localize in the islets of Langerhans, form deposits on β-cells, and enhance considerably the presentation of diabetogenic antigens. Thus, systemic recognition of a native circulating self-protein is sufficient for generation of autoantibodies that precipitate pathogenic outcomes in the target organ.
Role of the transcription factor Ikaros in development of autoimmune disease
Authors: Schjerven Hilde , Frietze Seth , Hai Si-Han , Hermiston Michelle , Kogan Scott , Muschen Markus 
Published: January 2023

Abstract
The etiology of autoimmune disease is not fully understood, but disease development is believed to be due to a combination of genetic predispositions and environmental factors. Ikaros is a transcription factor that is an important regulator of hematopoiesis and critical for development of B cells. Recent genome wide association studies have found that several autoimmune diseases are associated with SNPs annotated to the Ikaros gene (IKZF1), as well as the closely related Ikaros family member Aiolos (IKZF3). Mouse models of autoimmunity are important research tools, and we recently reported the generation of two new Ikaros-mutant mice, with targeted deletions of the exons encoding the DNA-binding zinc finger 1 (ZnF1) or ZnF4 (Schjerven et al., 2013). These deletions did not abolish B-cell development, but rather illuminated differential roles of Ikaros at different stages of early B-cell development. Interestingly, we found that one of these mouse strains displayed very high levels of serum Anti-Nuclear Antibodies (ANA), indicative of autoimmune disease. Furthermore, we found by ex vivo studies that while wt spleen B cells require a “second signal” such as CD40L in addition to BCR stimulation (by anti-IgM), Ikaros-mutant B cells became activated and proliferated upon BCR stimulation alone without any externally added “second signal”. We hypothesize that Ikaros is important to limit autoimmunity and prevent auto reactive B cells by setting a requirement for the “second signal” in adaptive immune-recognition. Results will be presented from ongoing investigation using in vivo analysis of the autoimmune-related phenotype, and ex vivo analysis of Ikaros-dependent molecular mechanisms underlying the response to stimuli in B cells.