bluuebunny/crossref_metadata_2025_split huggingface dataset:
Examining the Role of Neutrophils in Respiratory Syncytial Virus (RSV) Infection (168.26)
Authors: Stokes Kate , Moore Martin 
Published: January 2023

Abstract
The inflammatory process in the respiratory tracts of respiratory syncytial virus (RSV)-infected infants is not well characterized. Respiratory syncytial virus infects bronchial epithelial cells, resulting in epithelial activation and damage and an increased neutrophil response. However, the role of neutrophils in RSV pathogenesis is not known. To examine their role, we used a monoclonal antibody to Ly6G to deplete neutrophils in BALB/c mice infected with an RSV clinical isolate. Mice were then examined for weight loss, viral load, cytokine production, mucus production, and lung pathology. Administration of anti-Ly6G resulted in depletion of neutrophils from RSV-infected mice in blood and lungs. Neutrophil-depleted, RSV-infected animals lost more weight than infected isotype-control treated animals. There was, however, no significant effect on viral load. Neutrophil-depleted, RSV-infected mice produced less mucin than isotype-treated, RSV-infected mice. Neutrophil depletion during RSV infection also resulted in decreased TNF-α, IL-13, IL-4 and MIP-1α levels in the lung. Neutrophil depletion during RSV infection resulted in greater bronchiolar and alveolar edema, perivascular cuffing, alveolar hemorrhage, and vasculitis compared to control-treated, RSV-infected mice. This data shows that neutrophils play a major immune modulation role in RSV infection, promoting TH2 cytokine expression and airway mucus expression but also protecting against lung damage and disease severity.
Mass spectrometry-based global profiling of eicosonoid expression in response to influenza A virus infection (168.32)
Authors: Hilmer Kimberly , Graham Amy , Zickovich Julianne , Hilmer Jonathan , Obar Joshua 
Published: January 2023

Abstract
Understanding the innate inflammatory pathways activated during influenza A virus infection is vital to developing effective therapeutics. Our results demonstrate that the inflammatory cascades induced by different influenza A virus strains are not identical. Mice infected with A/WSN/33 show less airway hyper-reactivity than mice infected with A/PR/8/34, likely due to decreased expression of the IL-33/IL-13 axis of inflammation. In addition, we have examined the lipidomic expression profile during flu infections from each of these strains. Lipid extracts were prepared from whole lungs, and specific lipid mediators were both identified and quantified using UHPLC-MS. This approach provides an automated, high-throughput, and high-sensitivity technique to characterize the lipidomic fingerprint of the infection resulting from various influenza A strains. The compounds differentially expressed between strains will ultimately be used to develop targeted therapeutics to lessen both the morbidity and mortality of the seasonal flu.
RON Receptor Tyrosine Kinase, a Negative Regulator of Inflammation, is Decreased During SIV Associated Central Nervous System Disease. (168.33)
Authors: Cary Daniele , Clements Janice , Henderson Andrew 
Published: January 2023

Abstract
Expressed on tissue resident macrophages, the receptor tyrosine kinase, RON, functions to maintain inflammation homeostatisis by activating genes that promote wound repair and resolve inflammation; while repressing genes that perpetuate tissue damage. Chronic HIV infection is associated with dysregulated inflammation that contributes to organ diseases, and we hypothesize that altered RON expression contributes to the development of HIV associated CNS disease. We developed a tractable in vitro model using macrophages isolated from human tonsil; these cells express CD14, CD16, CD206, CCR5, and RON. Treatment with RON ligand induced arginase and inhibited HIV and IL-12p40. Following infection, RON expression was down-regulated. We utilized a well characterized SIV model in macaques to explore the temporal regulation of RON in the brain. Following prolonged SIV infection, RON expression decreased. RON was maintained in animals that did not develop CNS disease and was reduced in macaques that demonstrated CNS disease symptoms. Arginase expression was reduced during late infection whereas expression of the inflammatory genes, interleukin 12p40 and TNFα was elevated. Our data support a model in which during acute SIV infection in the brain, active inflammation is controlled by RON, however, following prolonged infection, RON expression is decreased, genes that quell inflammation are repressed, and inflammatory mediators are induced to directly promote CNS disease.
Inhibitory effects of calcitonin gene-related peptide (CGRP) on murine retroviral infection of glial cells (168.34)
Authors: Cao Ling , Malon Jennifer , Vaughn James , Birch Julie 
Published: January 2023

Abstract
We have observed that C57BL/6 (B6) mice infected with a murine retroviral isolate, LP-BM5 developed peripheral neuropathy, along with an elevated expression of calcitonin gene-related peptide (CGRP), in addition to glial activation in the spinal cord. To investigate how these spinal cord changes might contribute to the development of peripheral neuropathy, we examined the effects of CGRP on LP-BM5-induced glial cytokine responses in vitro. Mixed spinal cord glia were prepared from adult B6 mice and infected with various doses of LP-BM5 (0, 1, 10, 100, and 1000 pfu/ml), with or without CGRP co-treatment (1ug/ml). Resulting levels of selected cytokines (IL-1β, IL-6, TNFα, IFNγ, IL-12, and CCL2) in the culture supernatants were measured 1, 4, and 7 days post-treatment via ELISA. Viral loads in both the cells and supernatants were determined at day 7 post-treatment using plaque assay. While little virus was detected in the supernatants, CGRP treatment appeared to reduce viral loads in the infected glial cells. This reduction was correlated with the decrease of LP-BM5 induced TNFα production. IL-6 and CCL2 production, however, were enhanced by CGRP treatment. Further, CGRP did not affect the levels of IL-12p40, and IL-1β, IFNγ. IL-12p70 was not detectable following any treatment. Altogether, the neuropeptide CGRP appears to inhibit viral infection of glial cells. Further study will help to understand the role of CGRP during retroviral infection-induced neurological defects.
Varicella zoster virus can reactivate and infect cerebral adventital fibroblasts, suppressing Jak/STAT and causing stroke. (168.35)
Authors: James Stephanie , Choe Alexander , Traktinskiy Igor , Nagel Maria , Gilden Donald 
Published: January 2023

Abstract
Primary varicella zoster virus (VZV) infection causes varicella (chickenpox), after which virus becomes latent in ganglionic neurons. A declining cell-mediated immunity to VZV leads to virus reactivation, manifesting as zoster (shingles). Zoster is often complicated by chronic pain (post herpetic neuralgia) and other serious neurological disorders such as meningoencephalitis and myelitis. VZV vasculopathy results in transient ischemic attacks and stroke, and is the result of productive virus infection in cerebral arteries. More than 90% of the 6.75 billion world population harbors latent VZV. Adults with zoster have a 30% increased risk of stroke within the following year. Although the exact incidence of stroke caused by VZV is unknown, more than 900,000 people in the United States will develop zoster annually; thus, the magnitude of stroke caused by VZV is considerable. In a study of 3 VZV stroke patients, cerebral arteries were positive for VZV antigen and had intimal layer thickening. Previous experiments with VZV infected skin fibroblasts have shown that VZV inhibits the expression of STAT1α and Jak2, resulting in decreased expression of Interferon α and β (IFNα/β). Here we show that in VZV infected cerebral adventitial fibroblasts STAT1α and Jak2 are suppressed, which is not observed in controls. Thus, we postulate that viral infection of the adventitial fibroblasts suppresses the Jak/STAT pathway which allows for viral persistence, and potential stroke.
Pulmonary NK cell recruitment during influenza A virus infection (168.5)
Authors: Carlin Lindsey , Heusel Jonathan , Legge Kevin 
Published: January 2023

Abstract
Natural Killer (NK) cells are vital components of the antiviral immune response, but their contributions in defense against influenza A virus (IAV) are not well understood. To better understand the contribution of NK cells to IAV immunity, we examined the magnitude and kinetics of the NK cell response during high- and low-dose IAV infections. While an increased accumulation of NK cells in the lung in high-dose versus low-dose infections was observed, the NK cell response in the lymph nodes (LN) was blunted in high-dose infections. Using a dual labeling method to determine the rate of proliferation of pulmonary NK cells, our results show that while some local NK cell proliferation occurs during IAV infection, it is not enough to account for the increase in NK cells observed, suggesting that NK cells are instead largely recruited to the lungs. Therefore, we next determined the roles of the chemokine receptors CCR5 and CXCR3 in NK cell trafficking to the lung and LN using receptor blocking and knock out models. All together, our results demonstrate that NK cells are actively recruited to the lungs during IAV infection, suggesting differential NK cell recruitment could modulate the immune response to IAV during high- and low-dose IAV infections.
Role of lipoxygenase enzymes in RSV-induced differentiation of alternatively activated macrophages (AA-Mϕ) (168.7)
Authors: Shirey Kari Ann , Lai Wendy , Pletneva Lioubov , Blanco Jorge , Vogel Stefanie 
Published: January 2023

Abstract
Severe RSV-induced bronchiolitis has been associated with a mixed “Th1” and “Th2” cytokine storm. Arachidonic acid is the substrate for both COX-2 and 5-LO. In mice, TLR4, IFN-β, and IL-4/13 are required for RSV-induced differentiation of AA-Mϕ that counteract COX-2-induced lung pathology. We hypothesized that there is a reciprocal relationship between induction of COX-2 and AA-Mϕ genes during RSV infection. RSV infection of WT peritoneal macrophages resulted in the induction of 5-LO gene expression early in infection as well as activation of 5-LO enzyme activity later in infection. RSV-infection of 5-LO-/- macrophages and mice showed a loss of AA-Mϕ gene expression, while enhancing COX-2 mRNA and increasing the lung pathology in vivo. Blocking 5-LO or 15-LO with enzyme-specific inhibitors (AA-861 and PD-146176, respectively) also blocked differentiation of RSV-induced AA-Mϕ. RSV-infected cotton rats treated with saline or the COX-2-specific inhibitor, paracoxib, exhibited enhanced RSV-induced 5-LO mRNA expression. These data greatly extend our knowledge of the complex signaling that occurs during RSV infection as well as suggest novel treatment approaches for RSV that promote AA-Mϕ differentiation.
Viral induced encephalitis initiates distinct and functional CD103+ CD11b+ brain dendritic cell populations within the olfactory bulb. (168.8)
Authors: D'Agostino Paul , Kwak Changsoo , Vecchiarelli Haley , Gal Toth Judit , Miller James , Masheeb Zahrah , Bulloch Karen 
Published: January 2023

Abstract
Dendritic cells (DC) are antigen presenting cells found in both lymphoid and non-lymphoid organs, including the brain (bDC) of Cd11c/eyfp transgenic C57Bl/6 mice. Using an intranasal vesicular stomatitis virus infection, we demonstrated that EYFP+ cells amass in areas associated with viral antigens, take on an activated morphology, and project their processes into infected neuronal tissue within the olfactory bulb. These bDC separated into three EYFP+ CD45+ CD11b+ populations: EYFP+ CD45int CD11bhi (P1), EYFP+ CD45hi CD11bhi (P2) and EYFP+ CD45hi CD11bint (P3) populations. All but the P1 population were able to functionally promote both T lymphocyte proliferation and TH1 cytokine production using an OT transgenic antigen presentation assay. Using radiation chimeras, P1 was shown to emanate from the brain, while P2 was established to be peripherally derived. Interestingly, the P3 population was of indeterminate origin, being both radiosensitive and not replenished by donor bone marrow. Finally, each EYFP+ population contained CD11b+ CD103+ sub-populations, and could be distinguished in terms of M-CSFR, Gr-1, and Ly-6C expression. Such phenotypic analysis point to P1 as an activated sub-population of microglia, while P2 was more typical of mucosal DC subsets and P3 appeared to be similar to monocyte-derived DC lineages.
Altered human γδ peripheral blood lymphocyte homeostasis and impaired influenza A viral response in obesity (168.9)
Authors: Costanzo Anne , Taylor Kristen , Han Peggy , Fujioka Ken , Jameson Julie 
Published: January 2023

Abstract
Obesity is an independent risk factor for severe and fatal influenza infection. Little is known about how obesity impacts influenza-specific T cell responses in humans. In healthy adults, γδ T cells are the major circulating population of T cells that rapidly respond to influenza virus infected APCs. The majority express a Vγ9Vδ2 TCR that recognizes phosphoantigens expressed by infected or transformed cells. To determine whether this anti-viral T cell population was impacted by obesity, human peripheral blood mononuclear cells were isolated from non-obese and obese donors and analyzed for homeostatic T cell numbers and function. We found the percentage of Vγ9Vδ2 T cells negatively correlates with increased body mass index (BMI), suggesting the severity of obesity impacts γδ T cell homeostasis. Furthermore, Vγ9Vδ2 T cells exhibit a skewed maturation, with a higher propensity toward a unique γδ T effector CD45RA+ memory cell population. We have identified that obese patients show reduced expression of the homing molecule CCR5 on γδ T cells as well as reduced IFN-γ and TNF-α production in response to influenza infection. This blunted Vγ9Vδ2 T cell response in obese patients is T cell intrinsic as function can be rescued using a strong TCR agonist. Our data show that one arm of the antiviral immune response to influenza is compromised by obesity. These studies support the concept that therapies should be developed to restore immune homeostasis in obese patients.
Influenza A virus pathogenesis: Identification of host factors that contribute to severe respiratory distress (169.18)
Authors: Le Thuc-vy , Steel John , Compans Richard 
Published: January 2023

Abstract
The Center for Disease Control estimates that influenza virus infections contribute to 3,000-49,000 deaths in the United States annually. This broad range in deaths associated with influenza virus infections underscores the wide range in pathogenic effects that is observed following infection with influenza virus. While several factors contribute to pathogenesis, the mechanisms that govern how some infections result in severe respiratory distress, while others are resolved inconsequentially, are only partially defined. To explore the host cellular components that contribute to influenza virus pathogenesis, we utilized two genetically related strains of the 2009 H1N1 pandemic influenza virus A/Netherlands/602/2209 and A/California/07/2009. Although these viruses share nearly 100% sequence identity, mice that are infected with the different viruses exhibit great disparity in the severity of disease symptoms as measured by body weight, mortality and viral replication in the respiratory tracts. Our study reveals several key components that are critical in the balance between an immune response against influenza virus in which the virus is rapidly cleared and damage to the host is moderate, versus a response resulting in severe pathogenic effects that is marked by respiratory distress.
Regulatory Role of HMGB1 on complement activation via the classical pathway (169.3)
Authors: Kim Sook Young , Lee Sang Eun , Kwak Man Sup , Shin Jeon-Soo 
Published: January 2023

Abstract
Serum complement proteins play an important role in resistance to infection. The complement system is strongly activated in sepsis and the complement-activated products of anaphylatoxins have biological activities leading to inflammation. The nuclear protein, high mobility group box 1 protein (HMGB1) is released by inflammatory stimuli and plays a key role as a pro-inflammatory cytokine-like molecule. To investigate whether HMGB1 can induce complement activation, we have looked at the relationship between HMGB1 and complement system in this study. We first demonstrated the involvement of HMGB1 in binding to C1q for classical pathway activation. We observed the activation of complement cascade components such as C4b, C3b, C3c, and the membrane attack complex after C1q activation by HMGB1 in a dose-dependent manner. Moreover, complement activation could be observed in serum from mice after the intravenous injection of HMGB1. We then investigated the involvement of the A and B domain of HMGB1 during complement activation. HMGB1 B box, a well-known pro-inflammatory domain, was able to activate the complement system like wild type HMGB1, while HMGB1 A box showed no activation. The possible role of HMGB1 A box as an inhibitory domain in complement activation is now under investigation. In conclusion, the classical pathway of complement activation could be activated by the interaction of HMGB1 and C1q, showing the novel role of HMGB1 in the pathogenesis of HMGB1-induced shock.
Type I Interferon Regulates the Dynamics of Innate Myeloid Sentinel Cells During Persistent Viral Infection (169.7)
Authors: Nayak Debasis , Johnson Kory , Heydari Sara , Roth Theodore , Zynselmeyer Bernd , McGavern Dorian 
Published: January 2023

Abstract
Viral infections of brain usually trigger inflammatory responses that give rise to a wide range of pathological outcomes. To gain novel insights into how the brain responds to a virus as it establishes persistence, we utilized genomic and two-photon imaging approaches to study a pure innate immune response to a lymphocytic choriomeningitis virus (LCMV) infection. This was accomplished by monitoring temporal gene expression patterns and myeloid cells dynamics in the brain of CX3CR1-GFP mice lacking LCMV-specific T cells. Within the first two weeks of infection, brain resident myeloid cells were highly activated and responded directly by sequestering DsRed-labeled viral antigen. At the genomic level, this triggered massive changes in gene expression that were attenuated over time (likely due to inhibition by viral nucleoprotein) and were all directly linked to type I interferon (IFN-I) signaling. Surprisingly, in the absence of IFN-I signaling, no genes were differentially expressed in the nervous system despite the presence of persistent virus. In addition, two-photon imaging studies revealed that IFN-I receptor deficient innate immune sentinels were completely unresponsive to the establishment of a persistent viral infection. These data demonstrate that IFN-I drives all innate immune activity in the brain following LCMV infection, and deficiency in IFN-I signaling renders LCMV invisible in its murine host.
Regulatory T cells shape the effector and memory T cell response to virus infection in the tissues (170.15)
Authors: Graham Jessica , Lund Jennifer 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are well known for their role in dampening the immune responses to self-antigens and thereby helping to prevent autoimmune disease. However, they must also permit immune responses to occur against foreign infectious agents. Using a mouse model of West Nile Virus (WNV) infection, we examined Treg kinetics, trafficking, and activation at both acute and memory time points. We showed that Treg numbers increase in both the secondary lymphoid organs and CNS after infection, and become activated similar to effector T cells. Using the Foxp3DTR knock-in mice, we have performed experiments to study the course of WNV infection in the presence or absence of Tregs. We found that compared to ”normal” mice, Treg deficient mice had increased antigen-driven production of IFN-g from both CD4 and CD8 T cells in both the spleen and CNS. Additionally, Treg deficient mice had an increased frequency of poly-functional T cells. In mice lacking Tregs, there were greater numbers of short lived effector CD8+ cells at acute infection time points in both the spleen and brain. At a memory time point, there were significantly fewer antigen-specific T cells present in the CNS of mice deficient in Tregs at the time of infection compared to Treg sufficient mice. Studies are currently underway to examine the role of Tregs in CD4 memory using this Treg depletion model.
Analysis of early differential immunosuppressive patterns in the MAIDS model (170.23)
Authors: Akalu Yemsratch , Kwaa Abena , Sceats Lindsay , Stranford Sharon 
Published: January 2023

Abstract
Murine acquired immunodeficiency syndrome (MAIDS), induced by exposure to murine leukemia virus (MuLV), serves as an animal model to study human AIDS. While infection with the LP-BM5 isolate of MuLV leads to progressive immune compromise in MAIDS-susceptible C57BL/6 mice, resistant BALB/c mice launch directed and controlled immune responses to eradicate the virus and develop protective immunity. Since modulation of immunosuppressant molecule activities after viral infection may shape disease progression, we aimed to characterize key early differences in the expression of two natural immunosuppressive proteins, indoleamine 2, 3-dioxygenease (IDO) and interleukin-10 (IL-10). Preliminary experiments showed significantly higher IDO mRNA expression levels in the secondary lymphoid organs of naïve and infected disease-susceptible C57BL/6 mice than in resistant BALB/c mice. In our follow-up studies we used commercially available ELISA kit to measure IL-10 and developed our own ELISA-based assay to measure mouse-IDO. The findings indicate that expression of IDO protein is induced early after viral infection. The IL-10 ELISA indicated significant differences in IL-10 expression between the two strains of mice during the first week of infection. These results suggest that these immune mediators may preferentially activate immunosuppression pathways in MAIDS-susceptible mice.
Regulatory T cells can be expanded from HIV-1 infected individuals with preservation of suppressive function and TCR repertoire (170.26)
Authors: Angin Mathieu , King Melanie , Klarenbeek Paulus , Moodley Eshia , Rezai Ashley , Piechocka-Trocha Alicja , Toth Ildiko , Walker Bruce , Addo Marylyn 
Published: January 2023

Abstract
While natural regulatory T cells (Tregs) are being explored as therapeutic modalities in the context of autoimmune diseases and transplantation, their role in HIV-1 pathogenesis is less well defined and to date it is unknown if Tregs can be isolated and expanded from HIV-1 infected individuals. Tregs and conventional T cells (Tconvs) were isolated from the peripheral blood of HIV-1 infected and healthy donors using flow-based cell sorting, stimulated and cultured. Expanded Tregs were phenotyped by flow cytometry and qPCR. TCR repertoire diversity was investigated by high-throughput sequencing. Suppressive function was assessed using flow-based proliferation and chromium release cytotoxicity assays. Infectability of expanded Tregs was investigated using VSV-G pseudotyped HIV-GFP. Tregs from HIV-1+ individuals, including infants, were successfully expanded with a mean fold change of 29 at day 7. Expanded Tregs expressed high levels of FOXP3, CTLA4 and HELIOS and a demethylated TSDR (Treg-specific demethylated region). The TCR repertoire of expanded Tregs was not skewed and they maintained their suppressive function in vitro. Expanded Tregs were readily infectable with HIV-GFP. Our data show that Tregs from HIV-1 positive individuals can be successfully isolated and expanded ex vivo. This approach holds great potential for detailed studies on the role of regulatory T cells in the setting of HIV-1 infection and has implications for potential future therapeutic use of Tregs.