bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel mouse model of MCMV-induced adaptive NK cells
Authors: Jensen Isaac J , Martin Matthew D , Tripathy Sandeep K , Badovinac Vladimir P , Farber Donna L 
Published: January 2023

Abstract
NK cells are important mediators of viral control with the capacity to form adaptive immune features following infection. However, studies of infection-induced adaptive NK cells require adoptive cell transfer to lower the precursor frequency of ‘antigen-specific’ NK cells, potentially limiting the diversity of the NK cell response. In seeking an unmanipulated model to probe the adaptive NK cells, we interrogated a wide range of Collaborative Cross (CC) inbred mice, inbred mouse strains which exhibit broad genetic diversity across strains. Our assessment identified and validated a putative ‘ideal’ CC strain, CC006, which does not require manipulation to generate and maintain adaptive NK cells. Critically, CC006 mice, in contrast to C57Bl/6 mice, are capable of developing enhanced NK cell-mediated protective responses to MCMV infection following m157-mediated vaccination. This work both furthers our understanding of adaptive NK cells and demonstrates the utility of CC mice in the development and interrogation of immunologic models.
Supported by T32AI007511 (IJJ), T32AI007485 (IJJ), 5R01AI114543 (VPB), 1R35GM134880 (VPB), U01-AI150680 (DLF)
Liver Biopsies Reveal Temporal Changes in Pathology and Macrophage Function following SIV Infection of Rhesus Macaques
Authors: Derby Nina R , Fancher Katherine A , Biswas Sreya , Yusova Sofiya , Luevano-Santos Cristina , Smedley Jeremy , Pacheco Cristina , Burwitz Benjamin , Sodora Donald L 
Published: January 2023

Abstract
Non-alcoholic fatty liver disease (NAFLD) contributes to morbidity and mortality in HIV-infected individuals. To evaluate liver immune changes during SIV infection of macaques, we obtained liver biopsies laparoscopically at 2, 6, and 16 weeks post-infection as well as at necropsy (32 weeks). SIV infection was associated with liver dysfunction that included elevated liver enzymes during acute infection, higher liver weight at necropsy, and histologic changes within the liver lobule (functional unit of the liver) including sinusoidal dilatation near central veins and portal expansion. The type-1 interferon (IFN-1) response (MX1 expression) increased over time post-SIV infection, initially being observed in macrophages (week 2) with a switch to a mixed hepatocyte/monocyte/macrophage response by 16–32 weeks. Over the course of SIV infection, macrophage frequencies also increased – both infiltrating (CD68+CD163+CD206-) and resident (CD68+CD163+CD206+) tissue macrophages – and these macrophages were found more near central veins over time. In contrast, CD68+CD163-CD206− trafficking myeloid cells/monocytes were detected predominantly in the periportal zone of the lobule and associated with elevated MX1 expression in that zone. Our findings identify pathologic changes within the livers of SIV infected macaques as early as 2 weeks post-infection, IFN-1 responses that are initially observed in liver macrophages, and an accumulation of macrophages associated with signs of inflammation by week 32. These studies offer quantitative and spatial insights to inform macrophage-targeted therapeutic approaches that may improve liver function during HIV infection.
Supported by grants from NIH (R01AI134630)
Using single cell spatial and transcriptional analysis to understand early host response to RNA virus infections
Authors: Arakkal Leanne , Sturdevant Gail , Robertson Shelly , Meade-White Kimberley , Germain Ronald N , Best Sonja M , Speranza Emily 
Published: January 2023

Abstract
The RIG-I like receptor pathway, which signals through the mitochondrial anti-viral signaling (MAVS) protein, initiates the type I interferon (IFN) response to RNA viruses such as vesicular stomatitis virus (VSV) and Ebola Virus (EBOV). To investigate the mechanism of MAVS-mediated control of VSV and EBOV infections in vivo, we utilized single cell RNA sequencing and multiplex immunohistochemistry (mIHC) in C57Bl/6J and MAVS−/− animals to understand the cellular response spatially and transcriptomically in the draining lymph nodes (dLN). Identification of virally infected cells through scRNA-Seq allowed contrast of infected cells and bystander cells. Viral quantification of EBOV and VSV samples showed an increased viral load in MAVS−/− conditions and distinct patterns of dissemination through the LN. The relationship between viral load and IFN signaling revealed a negative correlation in EBOV infected cells with no relationship in VSV infected cells highlighting the importance of viral inhibition of the IFN response in EBOV infections. To assess these patterns spatially, mIHC antibody panels have been implemented on control and experimental tissue. An in-house algorithm for cell segmentation called RAPID was applied to a controlled experiment and revealed extensive batch effects from the staining process. Current work to perform batch normalization is being carried out to better assess changes in protein marker expression. Combining information gained from the sequencing analysis with the observed spatial organization through mIHC, we aim to develop a model illustrating the role of the MAVS-dependent pathway in controlling early virus replication.
Supported by Intramural research program of the NIH
Corticosteroid Treatment Results in Enhanced susceptibility to Influenza-Associated Aspergillosis
Authors: Worthley Devyn , Lupfer Christopher R 
Published: January 2023

Abstract
Coinfections that occur during an influenza A virus (IAV) infection are often severe and life threatening. Since the 1918 influenza pandemic, researchers and clinicians have recognized that infection with IAV predisposes individuals to a more severe form of pneumonia resulting from secondary bacterial infections. However, coinfection following IAV infection can occur with other pathogens including other viruses and fungal pathogens. In fact, over the last two decades, numerous case reports demonstrate that influenza coinfections with Aspergillus fumigatus deserves more scrutiny as a potential source of severe respiratory disease. Here, we report that mice with Influenza-Associated Aspergillosis (IAA) develop a mild form of disease unless concomitantly treated with corticosteroids. Importantly, corticosteroid usage impairs immune function resulting in enhanced fungal growth associated with altered commensal growth in the respiratory tract. Our findings agree with case reports that suggest corticosteroids may be involved in the development of IAA.
Supported by grants from Missouri State University Graduate College.
IL-15 synergizes with IL-12 to enhance NK cell function in vivo during chronic SIV infection
Authors: Babu Hemalatha , Govindaraj Sakthivel , Wagoner Isabelle , Ali Syed A , Ibebgu Chris , Amara Rama Rao , Villinger Francois , Velu Vijayakumar 
Published: January 2023

Abstract
Natural killer (NK) cells play a critical role in viral immunity. During early HIV/SIV infection, they are known to protect/limit viremia at the very early stage of infection. However, during chronic HIV infection NK cells are less functional, thus elicitation of highly functional NK cells are important for HIV cure strategy. It has been shown that cytokines IL-15 and IL-12 individually can activate NK cells during SIV infection. However, the synergistic effect of these cytokines to induce NK cell function has not been studied.
We performed in vivo experiment to evaluate the effect of cytokines IL-15 (20ug/Kg)/IL-15R(100ug/kg) and IL-12 (10ug/kg) and its combination in inducing functional NK cells during chronic infection (6 weeks PI) and with no cytokine treatment as a control. Lymphocytes from blood and lymph nodes were analyzed for NK cell functions using multicolor flow cytometry.
Our in vivo data clearly indicates that IL-15/IL-15R synergistically act with IL-12 to induce proliferation (Ki-67) and cytotoxicity (Granzyme-B+) of NK cells both in blood and lymph node. Interestingly, the cytotoxic NK cells CD16+ (FCgRIII) expanded significantly in the combination group. In addition, the degranulation capacity of NK cells (IFN-gamma+ CD107a+) significantly enhanced in the combination group when compared to either cytokine group alone, suggesting that combined cytokine treatment enhanced NK cell cytotoxicity during chronic SIV infection.
Our data clearly showed that the combination of IL-15 plus IL-12 cytokine treatment increased NK cell function in vivo with proliferative and cytotoxic capacity, which is important for HIV cure strategy considering directing functional NK cells to immune privileged sites such as lymph nodes.
This work was supported in part by National Institutes of Health Grants CFAR R03, R01 HD095741-01, 1R01AI148377-01A1 (to VV), U19 AI109633 (to R.R.A.), Emory University CFAR grant P30 AI050409, NIH Office of Research Infrastructure Programs (ORIP) grants P51 OD011132 and U42 OD011023 (to YNPRC) and R24 OD010947 to the Resource for NHP Immune Reagents.
Pre-existing atopy prevents mortality from respiratory viral infection via induction of neuregulin-1
Authors: Hussain Syed-Rehan A , Rohlfing Michelle , Santoro Jennifer , Resiliac Jenny , Grayson Mitchell H 
Published: January 2023

Abstract
Respiratory viral infections are associated with significant morbidity and mortality. RNA viruses, such as Respiratory Syncytial Virus (RSV) have been implicated in the development of asthma. We have a high-fidelity model of respiratory viral infection. Our mouse model utilizes a natural rodent pathogen, Sendai virus (SeV) similar to RSV, which faithfully replicates in mouse airway epithelial cells. We have shown that making these mice atopic (i.e, sensitized and challenged with house dust mite extracts) before viral infection prevents development of disease and more importantly prevents mice from dying of a normally lethal dose of SeV. Further, lungs and bronchoalveolar lavage fluid of atopic mice have significant increased levels of Neuregulin-1 (NRG1), potentially due to high expression of Nrg1 in dendritic cells as determined by RNAseq of CD11c+ cells. To determine a temporal relationship between NRG1 expression and respiratory viral infection, we utilized an in vitro system of well-differentiated human bronchial epithelial cells and mouse tracheal epithelial cells. Cells were treated on the basolateral side with NRG1 (10 to 100ng) for six-days before 4000 pfu of RSV-GFP or SeV-GFP inoculation, respectively. There was marked reduction in viral spreading (quantified by reduced GFP expression) with NRG1 treatment. Interleukin-6 (IL6) expression was reduced in NRG1 treated cells at 48h PI, as well. IL6 has been implicated with cellular stress and is involved in disease pathology. Therefore, reducing IL6 might affect the clinical presentation of serious infections. In conclusion our studies show that atopy induced NRG1 plays a novel role with a therapeutic potential in severe respiratory viral infections.
Supported by Abigail Wexner Research Institute at Nationwide Children’s Hospital, Start-up fund to MHG
Nanoparticles divert monocytes away from the lungs to improve outcomes after influenza virus infection in aged mice
Authors: Elhofy Adam , Kelley William J , Chen Judy , Murthy Tushar , Xu Qichen , Boyne Michael , Goldstein Daniel 
Published: January 2023

Abstract
Older people exhibit dysregulated innate immunity to respiratory viral infections, including influenza and SARS-CoV-2, to increase morbidity and mortality. Nanoparticles are a potential practical therapeutic that could divert exaggerated innate immune responses away from the lungs during viral infection. However, such therapeutics have not been examined for effectiveness during respiratory viral infection, particular in aged hosts. Here, we employed a highly lethal model of influenza viral infection in vulnerable aged mice to examine the ability of biodegradable, cargo-free nanoparticles, designated ONP-302, to resolve innate immune dysfunction and improve outcomes during infection. We administered ONP-302 via intravenous injection to aged mice at day 3 post-infection when the hyperinflammatory innate immune response is already established. We found that ONP-302 treatment diverted tissue-damaging inflammatory monocytes from the lungs to the spleen without impacting viral clearance or reducing chemo-attractive signals in the infected lung. Importantly, cargo-free nanoparticles reduced lung damage and histological lung inflammation and improved gas exchange and, ultimately, the clinical outcomes in influenza-infected aged mice. ONP-302 improves outcomes in influenza-infected, vulnerable, aged mice. Thus, our study provides vital fundamental information concerning a practical therapeutic which, if translated clinically, could improve disease outcomes for vulnerable older patients suffering from respiratory viral infections.
Supported by COUR Pharmaceuticals
Controlling herpetic inflammatory lesions by inhibiting glucose metabolism- on the different outcome of using 2DG and metformin
Authors: Berber Engin , Rouse Barry T 
Published: January 2023

Abstract
Ocular infection with herpes simplex virus-1 (HSV-1) results in a blinding immunoinflammatory lesion. Control can be achieved by inhibiting the metabolism of cells principally involved in lesion development. Inhibiting glucose metabolism with 2DG achieves this effect but if therapy is used when replicating virus is still present, herpes simplex encephalitis (HSE) is likely to ensue. We report that using metformin to inhibit glucose metabolism can also minimize herpetic ocular lesions and this occurs without the usual complication of HSE. We try to provide a mechanistic explanation for the different outcomes. We show that the innate cell response in corneal lesions was similarly diminished by both metabolic therapies, but the effects on the composition of cellular T responses differed. Thus 2DG had more inhibitory effects than metformin on CD4 Th1 T cells, the main orchestrators of corneal lesions as well as on Foxp3+ Treg, cells responsible for controlling lesions. Metformin therapy had minimal inhibitory effects on Treg such that the representation of Treg markedly increased in comparison to Th1 cells. The responses occurring in the trigeminal ganglia, the site to which HSV spreads and then sets up permanent latency, was also investigated. The nature of the inflammatory responses at the site differed in untreated and animals treated with 2DG or metformin. Thus, both therapies resulted in diminished inflammatory responses particularly the innate cell composition. The inhibitory effects of 2DG against the CD8 content of the inflammatory reaction (cells advocated as involved in containing latency and stopping virus from disseminating to the brain) was significantly greater than was the metformin therapy.
Supported by grants from NIH (R21AI142862 and R01EY005093)
Role of exosomal BTLA/HVEM signaling in HTLV-1 associated diseases and associated T-cell dysfunction.
Authors: Joseph Julie , Rao Abhishek , Connors Jennifer , Stoffel Victoria , Nootenboom Julianne , Rahmani Benjamin , Haddad Elias , Carey Alison , Jain Pooja 
Published: January 2023

Abstract
Immune checkpoint (ICP) mediators play pivotal roles in regulating a broad spectrum of immune responses against cancer and infectious diseases. In previous studies we have established the role of negative checkpoint receptors (NCRs, PD-1, TIGIT, LAG-3, etc.) in determining the quality of anti-HTLV-1 cytolytic (CTL) response in controlling proviral load and maintaining a asymptomatic state in majority of infected individuals. Interestingly, ICPs can be released in the soluble form and carried on the surface of small extracellular vesicles (50–200 nm) known as exosomes that possess immunomodulatory activity. Consequently, we profiled ICPs in isolated exosomes and culture media of HTLV-1 cell lines representing both HAM/TSP and ATLL. High levels of BTLA (B-and T-Lymphocyte Attenuator) and its ligand HVEM (Herpes virus entry mediator) along with PD-1 (Programmed cell death receptor-1) and its ligand PD-L2 (not PD-L1) were observed in soluble and exosomal forms as well in the sera of HAM patients that carry high proviral load and viral proteins such as Tax. HTLV-1 infection, Tax protein and/or IFNy could serve as causative mechanism for increased ICP levels in HAM patients. Indeed, treatment with anti-retroviral drugs significantly reduced ICP levels confirming specificity of our observations. Functionality of exosomal BTLA/HVEM is being validated in the ongoing studies along with their direct role in regulating CD8 T-cell functions and cytolytic potential upon blockade to provide a novel therapeutic target for HAM/TSP and other neuroinflammatory diseases.
Supported by grants from NIH: 1R01NS0971-47, T32-MH079785
Radioresistant cells in STING gain-of-function mice initiate lymphocyte dependent lung inflammation and IFNγ dependent mortality
Authors: Gao Kevin MingJie , Marshak-Rothstein Ann , Fitzgerald Katherine A 
Published: January 2023

Abstract
Pediatric patients with gain-of-function (GOF) mutations in the cytosolic dsDNA sensing adaptor STING develop an autoinflammatory syndrome known as STING Associated Vasculopathy with onset in Infancy (SAVI). SAVI patients show elevated interferon signature genes and suffer from interstitial lung disease (ILD) with lymphocyte predominate bronchus associated lymphoid tissue (BALT). We genetically engineered SAVI mice (STING V154M or VM) that recapitulates human disease, including lymphocyte rich ILD, and found by genetic ablation that lymphocytes are required for lung pathology. However, while ablation of either T or B lymphocytes prolongs survival, lung immune aggregates persist, indicating either can independently be recruited as BALT. Using radiation chimeras, we found that hematopoietic stem cell (HSC) derived STING GOF was neither sufficient nor required for VM lung disease, suggesting a role for lung radioresistant parenchymal and stromal cell STING GOF in SAVI. We identified lung endothelia as putative radioresistant cells which express STING and confirmed expression by IF microscopy and FACS. FACS analysis also revealed SAVI lung endothelia upregulate MHCII, but only when sufficient for IFNgR. VM T cells produce IFNg, and IFNgR deficiency was sufficient to prevent endothelial MHCII upregulation and attenuate mortality. However, HSC STING GOF was not required for endothelial MHCII upregulation, suggesting stromal intrinsic STING GOF initiates this outcome. These findings suggest that activation of STING in stromal and parenchymal cells play a key role the initiation of lung inflammation in patients with connective tissue disease ILD.
Supported by: T32 GM107000 (MSTP); T32 AI132152 (KMG); F30 HL154674 (KMG); AI128358 (AMR/KAF); Lupus Research Alliance
The role of NKG2D signaling in NOD diabetes is affected by the microbiota
Authors: Markiewicz Mary A , Bedrosian Zoe K , Ruark Elizabeth , Krausz Kelsey 
Published: January 2023

Abstract
The drivers of type 1 diabetes are islet-specific T cells. Understanding the mechanisms by which these cells are activated is critical to developing better strategies to inhibit the disease. Signaling through NKG2D is implicated in this process, but its role is unclear owing to conflicting results from studies performed to date. Using non-obese diabetic (NOD) mice genetically deficient in NKG2D, we demonstrate that the effect of NKG2D signaling on NOD diabetes varies with housing conditions, with NKG2D signaling having the greatest role in germ-free mice. We show this signaling does not occur through interaction with ligands expressed within pancreatic islets, as we demonstrate that NKG2D signaling within the islets instead inhibits NOD diabetes development. Specifically, we show that NOD mice we generated with a genetic deficiency in the endogenous NKG2D ligand expressed in the islets, H60a, have increased diabetes development, whereas H60a-deficient NOD mice expressing the NKG2D ligand RAE1ɛ in β-islet cells (via a transgene) have reduced diabetes. Further, unlike the decrease in diabetes observed in germ-free NKG2D-deficient mice, there was no difference in diabetes development between germ-free H60a-deficient and wild-type NOD mice. Taken together, these data demonstrate that (1) the composition of the microbiota affects how significant a role NKG2D signaling plays in promoting NOD diabetes and (2) this signaling occurs outside of the pancreatic islets. We further demonstrate that antigen presenting cells (APCs) in the pancreatic lymph nodes of both specific pathogen free (SPF) and germ-free NOD mice express members of the NKG2D ligand family RAE1, indicating this may be the site of the diabetes-promoting NKG2D signaling.
Supported by grants from DOD (PR181543), Emilie Rosebud Diabetes Research Foundation, KUMC Diabetes Institute, and Beatson Foundation (2020-004). In addition, the University of Kansas Medical Center Flow Core was utilized, which was funded in part by NIH (P30 GM122731; P30 GM103326; P30 CA168524).
Treatment with TLR7/8 agonist compromises intestinal epithelium integrity in a lupus prone mouse model.
Authors: Terrell Morgan Lee , Elshikha Ahmed Samir , Kanda Nathalie , Brown Josephine , Zeumer-Spataro Leilani , Morel Laurence 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies leading to chronic inflammation and subsequent organ damage. Both spontaneous and induced mouse models have been central to the dissection of disease mechanisms. Mounting evidence suggests that increased gut permeability, or leaky gut, and the resulting translocation of pathobionts or their metabolites can impact SLE pathogenesis. Resiquimod (R848), a TLR7/8 agonist has been shown to induce lupus-like autoimmunity in various non-autoimmune mouse strains. Here, we examined the effect of the epicutaneous administration of R848 in the B6.Sle1.Sle2.Sle3 triple congenic (TC) spontaneous mouse model of lupus as compared to their non-autoimmune B6 controls. R848 induced a much stronger autoimmune activation in pre-autoimmune TC mice than in B6 controls. Unexpectedly, R848 decreased gut barrier integrity in TC but not in B6 mice. Immunofluorescence staining of the ileum showed a reduced expression of the tight junction protein claudin-1 and an increased infiltration of immune cells in R848-treated TC as compared to either R848-treated B6 or untreated control mice. The reduced intestinal epithelial integrity was functionally confirmed with a FITC-dextran intestinal permeability assay. These results demonstrate for the first time that TLR7/8 activation, which plays an essential role in autoimmune activation, also induces a leaky gut in lupus-prone mice. Ongoing studies are expected to unveil the causal relationships between TLR7/8 activation, gut barrier integrity, and microbial dysbiosis in SLE.
Supported by R01 AI143313 from the NIH to LM