bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anti-inflammatory effects of PUFA’s on T cells reduce TCR affinity for antigen
Authors: Kolawole Elizabeth Motunrayo , Evavold Brian D. 
Published: January 2023

Abstract
Polyunsaturated fatty acids (PUFAs) have been shown to modulate inflammation yet little is known on their specific effects on recognition of antigen by T cells despite. Since the T cell receptor (TCR) and its peptide-MHC (pMHC) antigenic ligand are embedded in the membrane, incorporation of omega-3 fatty acids into lipid membranes could likely change the 2D kinetics of TCR:pMHC engagement and the membrane environment. We used the 2D micropipette adhesion frequency assay to define the proximal kinetics of antigen recognition by T cells on a single cell basis. We found that dietary fish oil modifies the structural lipid content in plasma membranes as measured by mass spectrometry, resulting in reduced TCR affinity for antigen at the clonal and polyclonal level. The reduced affinity led to deceased frequency and effector functions from both CD4 and CD8 T cells at the peak immune response to viral (LCMV) infection. Thus, our work provides mechanistic insight for the anti-inflammatory effects of PUFAs by altering plasma membrane composition which leads to decreased 2D kinetics of antigen recognition in T cells and dampened response.
Z-DNA binding protein 1 (ZBP1) is an innate sensor of Influenza vRNPs and activates programmed cell death and assembly of NLRP3 inflammasome
Authors: Sannula Kesavardana , Kuriakose Teneema , Samir Parimal , Malireddi RK Subbarao , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
Influenza A virus (IAV) infection triggers pulmonary epithelial cell death and inflammation which promote bronchioalveolar tissue damage and failure of lung function. However, the mechanism of IAV induced cell death and inflammation remains unclear. RIP homotypic interaction motif (RHIM) has evolved to play critical role in cell death and inflammation. Z-DNA binding protein 1 (ZBP1/DAI) is unique among RHIM-containing proteins since it has additional Za nucleic acid binding domains. Although ZBP1 domain architecture suggests its function as nucleic acid sensor, its precise physiological role is not established.
We discovered that ZBP1 is a central activator of IAV induced programmed cell death and NLRP3 inflammasome activation in both immune and non-immune cells. Mechanistically, ZBP1 senses ribonucleoprotein complexes (vRNPs) of IAV and engage RIPK3-caspase-8 dependent apoptosis, necroptosis and NLRP3 inflammasome assembly. ZBP1 also drives RIPK1 dependent secretion of TNF and IL-6 cytokines after IAV infection. We also discovered that apical activation of RIG-I/MAVS/IFNAR signaling licenses ZBP1 upregulation and activation. In contrast to other RNA viruses, IAV replicates inside the nucleus. Accordingly, we observed localization of ZBP1 in the nucleus suggesting its role in sensing IAV inside the nucleus. Strikingly, we also found that ZBP1 undergoes ubiquitination in response to IAV infection to assemble cell death signaling complexes. In IAV infected mice, ZBP1 induces cell death in lung epithelial cells and its absence conferred better survival advantage, despite high viral titers in the lung. Overall, our work demonstrates the central role of ZBP1 in sensing IAV infection to trigger cell death and inflammation.
Natural killer cell regulation of HIV vaccine responses
Authors: Rydyznski Carolyn E. , McNally Jonathan P. , Gyurova Ivayla E. , Cranert Stacey A. , Waggoner Stephen N. 
Published: January 2023

Abstract
Understanding mechanisms that regulate humoral responses are crucial for the design of more efficacious vaccines. We have previously shown that natural killer (NK) cells suppress antiviral T-cell function and antibody production during acute virus infection in mice. NK cells also inhibited follicular helper (TFH) T-cell and germinal center (GC) B-cell responses following immunization with an alum-adjuvanted, non-replicating hapten-protein, NP-KLH. In addition, the frequency and type of mutations observed during SHM in NP-specific GC B cells is linked to NK cells, demonstrating that NK cells are important modulators of not just the quantity, but also the quality of the humoral response. To test the impact of NK cells on clinically-relevant HIV vaccine regimens, mice were depleted of NK cells prior to administration of an HIV envelope-expressing vaccinia virus vector. When NK cells were absent during priming, vaccine-induced TFH and GC B cell responses were increased along with production of HIV-specific immunoglobulin. Using a prime-boost vaccination scheme, we observed that NK cells also suppressed recall responses of B cells, such that combined depletion during both prime and boost immunizations had an enhanced effect on humoral responses. These results demonstrate that NK cells are important inhibitory regulators of affinity maturation and HIV-specific humoral responses, such that NK cells may limit vaccine-mediated generation of broadly neutralizing antibodies by curtailing GC responses. These data demonstrate the exciting potential to manipulate NK cells in order to promote effective vaccine-elicited protection against HIV infection.
Dock8-Positive CD4 T cell as Autoantibody-Inducing CD4 T (aiCD4 T) Cell That Causes Systemic Lupus Erythematosus (SLE): Proof of Concept of Self-Organized Criticality Theory as the Cause of SLE
Authors: Shiozawa Shunichi , Tsumiyama Ken , Miyazaki Yumi , Sakurai Keiichi , Miyazawa Masaaki 
Published: January 2023

Abstract

INTRODUCTION
We found in reproducible experiments in which the mice not prone to autoimmune disease were immunized repeatedly with antigen that overstimulation of CD4 T cells led to the development of aiCD4 T cell which had undergone TCR revision capable of inducing varieties of autoantibody and antigen-specific CTL via antigen cross-presentation, after which they caused SLE (Tsumiyama K et al., 2009). Here we identify aiCD4 T cell as DOCK8+CD4 T cell.

METHODS
aiCD4 T cell was searched by transferring different fractions of CD4 T cells of x12 OVA-immunized BALB/c female mice into naïve mice and tested if autoantibodies were raised, mass spectrometry, and FACS.

RESULTS
We focused CD45RBlo CD122lo PD1+CD4 T cell as aiCD4 T candidate, and its membrane uniquely expressed DOCK8. DOCK8+CD4 T cell was a large effector CD4 T cell with abundant ER and mitochondria, ICOS+ CXCR5-PD1+ Ly6C+ LFA1hi, produced increased IFNγ, IL4, IL6, IL17 and IL21, and its TCR repertoire was deviated. Upon transfer to naïve mice or to the x8 OVA-pre-immunized and CD4 T-depleted mice in which CTL was yet immature, DOCK8+CD4 T cells induced SLE, where anti-dsDNA/Sm Ab, glomerulonephritis of WHO IV/V types, skin liquefaction degeneration, splenic periarteriolar fibrosis with amyloid-like deposits classical of Onion-skin lesion, pericholangitis, pneumonitis, thyroiditis, perineuritis and panniculitis developed. Manifestations were cured by anti-DOCK8 Ab treatment.

CONCLUSION
We prove the self-organized criticality theory explaining that autoimmunity arises as a natural consequence of routine but exaggerated immune response against antigen when stimulated maximally beyond immune system’s self-organized criticality, and show that DOCK8+CD4 T cell causes SLE.
TFH-like cells promote TH17-induced neuro-inflammation
Authors: Quinn James L. , Kumar Gaurav , Agasing Agnieshka , Ko Rose M. , Axtell Robert C. 
Published: January 2023

Abstract
The cooperative role of T cells and B cells is a topical research area in multiple sclerosis. Previous studies, using experimental autoimmune encephalomyelitis (EAE), have shown that TH17 responses are correlated with ectopic B cell follicles in the CNS. As follicular T helper (TFH) cells control B cell responses, this study examines the role TFH cells play TH17-induced EAE. As previously reported, we found that B cells are a major cell type infiltrating the CNS of mice with TH17-EAE. We now show that the severity TH17-EAE was diminished in B cell deficient mice demonstrating that B cells contribute to the severity of disease in these mice. We next focused on the role TFH cells play in TH17-EAE. We found substantial numbers of TFH cells in the CNS of TH17-EAE mice and these cells are correlated with infiltrating B cells. Using transfer experiments, we found that TFH cells alone do not infiltrate the CNS but require the infiltration of TH17 cells to gain entry to the CNS. We next tested the effects of blocking TFH trafficking in TH17-EAE using an antagonistic antibody against CXCL13, the chemokine that binds to CXCR5 on TFH cells. We found that anti-CXCL13 treatment significantly reduced TH17-EAE disease. This treatment blocked T cells from entering the CNS but had no effect on B cells. Surprisingly, we observed that anti-CXCL13 treatment reduced disease in B cell-deficient mice. Collectively, these data demonstrate that B cells and TFH cells contribute to the severity of TH17-induced EAE. Furthermore, this report describes an important role for TFH cells that is independent of their support for B cell function and begins to answer important questions regarding their contribution in neuro-inflammatory diseases.
Neuroantigen-specific CD8 T cells inhibit ongoing demyelinating disease by autoregulating CD4 T cell responses using temporally distinct IFNγ- and perforin-dependent mechanisms.
Authors: Boyden Alexander , Brate Ashley A. , Karandikar Nitin 
Published: January 2023

Abstract
Pathogenesis of immune-mediated demyelinating diseases like multiple sclerosis (MS) is thought to be governed by a complex cellular interplay between immunopathological and immunoregulatory responses. We have previously shown that neuroantigen-specific CD8 T cells have an unexpected protective role in the mouse model of MS, experimental autoimmune encephalomyelitis (EAE). In this study, we interrogated the immunotherapeutic potential of PLP178–191-specific CD8 T cells. Here, we show that PLP178–191-specific CD8 T cells, when administered post-disease onset, swiftly ameliorated EAE progression and suppressed PLP178–191-specific CD4 T cell responses as measured by delayed-type hypersensitivity (DTH). To accomplish DTH suppression, PLP178–191-specific CD8 T cells required differential production of perforin and IFNγ. Perforin was ancillary for the rapid suppressive action of PLP178–191-specific CD8 T cells, but was necessary for maintenance of optimal longer term DTH suppression. Conversely, IFNγ production by PLP178–191-specific CD8 T cells was necessary for swift DTH suppression, but was less significant for maintenance of longer term suppression. These data indicate that neuroantigen-specific CD8 T cells employ an ordered regulatory mechanism program over a number of days in vivo during demyelinating disease and have mechanistic implications for this immunotherapeutic approach.
NKG2D signaling within the pancreas enhances CD8+ central memory T cell formation and decreases non-obese diabetic (NOD) diabetes development
Authors: Trembath Andrew P. , Sharma Neekun , Markiewicz Mary A. 
Published: January 2023

Abstract
The immune receptor NKG2D is implicated in autoimmune diabetes. However, the mechanism by which NKG2D signaling affects diabetes is unclear. Among the aspects not known are 1) the relevant anatomical site(s) of NKG2D signaling, 2) the cells in which NKG2D signaling occurs and 3) whether NKG2D signaling enhances or inhibits diabetogenesis. These are among the critical questions that must be answered to determine whether modulation of NKG2D signaling could be used in intervention strategies in type 1 diabetes. To determine whether NKG2D signaling within the pancreatic islets modulates autoimmune diabetes, we generated a novel NOD mouse strain in which the mouse NKG2D ligand RAE1-e is constitutively expressed in the beta-islet cells (RIP-RAE1e mice). Insulitis progressed faster in RIP-RAE1e mice compared with control NOD mice. Despite this, there was a significant decrease in spontaneous diabetes in RIP-RAE1e mice compared with control mice. Using flow cytometry, we compared the immune cells present in the pancreas and spleen of pre-diabetic RIP-RAE1e and control NOD mice. Similar percentages and numbers of CD4+ T cells, CD8+ T cells and NK cells were detected. Similarly, equivalent numbers of CD69+ CD4+T and CD69+CD8+T cells were present. However, RIP-RAE1e mice contained increased numbers of CD44hiCD62Lhi (central memory T cell, Tcm, phenotype) CD8+ T cells and decreased numbers of CD44hiCD62Llo (effector/effector memory, Tem, phenotype) CD8+ T cells in both the pancreas and spleen. This increased Tcm:Teff/em ratio was not observed in the CD4+ T cell compartment. Together, these data demonstrate that NKG2D signaling within the pancreas drives an increased ratio of CD8+ Tcm:Teff/em cells and is protective against NOD diabetes.
Autoreactive T cells use IFNγ and Fas Ligand to clear melanocytes in vitiligo
Authors: Strassner James P. , Azzolino Vincent N. , Richmond Jillian M. , Harris John E. 
Published: January 2023

Abstract
Vitiligo is an autoimmune disease of the skin in which melanocytes are selectively destroyed, leading to a disfiguring depigmentation. Studies by our group and others demonstrated that IFNγ-induced chemokines drive pathogenesis through the recruitment of CD8+ T cells that kill melanocytes. We examined potential mechanisms of melanocyte killing by T cells in our mouse model by performing adoptive transfer of autoreactive cells lacking IFNγ, perforin, or Fas ligand. We found that disease severity and T cell accumulation in the skin was reduced in mice that received either IFNγ KO or Fas ligand KO melanocyte-specific T cells; however, perforin was not required. We partially rescued the disease phenotype by performing 50:50 co-injections of effector molecule KO T cells with WT T cells, however KO T cells outcompeted WT cells in recruitment to the epidermis, suggesting IFNγ and Fas Ligand are needed in situ to cause disease. To determine if IFNγ signaling was directly cytotoxic to melanocytes, we used a Cre-Lox system in which the IFNγR was specifically deleted in melanocytes. Floxed/Floxed Cre+ littermates were not protected from disease, suggesting IFNγ signaling in melanocytes is dispensable for melanocyte clearance. In summary, IFNγ has pleiotropic effects in vitiligo, required both for T cell recruitment and functional killing of melanocytes. This further supports targeting IFNγ signaling for the treatment of vitiligo. In addition to IFNγ, Fas ligand is a new potential target for therapy that warrants further investigation.
Type-1 interferon receptor knockout mice generate high titers of ADCC antibodies in response to vaccination with ΔgD-2 but display defects in executing ADCC
Authors: Dardick Joseph , Weiss Kayla , Jacobs William R. 
Published: January 2023

Abstract
ΔgD-2 is a live-attenuated Herpes Simplex Virus type-2 (HSV-2) strain genetically deleted for glycoprotein D (gD) that elicits complete protection against HSV-1 and HSV-2 through antibody-dependent cell-mediated cytotoxicity and phagocytosis (ADCC and ADCP). The mechanisms by which immunogens elicit ADCC/ADCP activating antibodies are not well understood but type-1 interferons (IFNα/β) have been shown to play a role in activating humoral immunity and FcγR-mediated immune modulation. To investigate the role of IFNα/β in generating ADCC/ADCP antibodies, we vaccinated IFNα/β receptor knockout mice (IFNAR−/−−) with ΔgD-2 and compared the resulting immunity to that of WT mice. IFNAR−/− mice vaccinated with ΔgD-2 generated similar overall and isotype specific anti-HSV IgG titers to WT mice. Serum from vaccinated IFNAR−/− mice also induced significantly more ADCC/ADCP killing than serum from vaccinated WT mice (p&lt;0.05). To determine whether this serum initiated ADCC/ADCP in vivo, serum from vaccinated IFNAR−/− mice was passively transferred into naive WT and FcγRIV−/− mice. WT mice that received the vaccinated serum were partially protected from 10x LD90 challenge with HSV-2 but FcγRIV−/− mice were not (p&lt;0.05). Additionally, serum from vaccinated WT mice did not confer any protection to naive IFNAR−/− mice, indicating difficulty in mobilizing ADCC/ADCP (p&lt;0.01). However, IFNAR−/− mice that were vaccinated with ΔgD-2 were completely protected from morbidity and mortality following a 10x LD90 challenge with HSV-2 (p&lt;0.0001). Together, our data show that the production of ADCC/ADCP antibodies in response to ΔgD-2 is independent of IFNα/β, but IFNAR signaling is important for executing ADCC/ADCP protection.
Quantitative Bead-Based Multiplex Immunoassays for Mouse Anti-Virus Immune Response
Authors: Zhao Amy , Sun Binggang , Lehmann Jason , Ji Shaoquan , Jiang Weiping 
Published: January 2023

Abstract
In response to pathogens, especially viruses, cells release interferons and other cytokines to combat infection. Interferons are typically divided into three types: 1 (e.g., IFN-α, IFN-β), 2 (e.g., IFN-γ), and 3 (e.g., IFN-λ1, IFN-λ2). All inter-ferons are important for fighting viral infections and for regulating the immune system. In addition, interferons are critically involved in cancer and autoim-mune diseases such as psoriasis, systemic lupus erythematosus, and multiple sclerosis. Studying the expression profile of interferons and other related cyto-kines is essential to understanding the immune response against viral pathogens and other related disease processes.
We have developed a multiplex panel, using fluorescence-encoded beads, which is suitable for use on commonly available flow cytometers. The panel simultaneously quantifies 13 mouse anti-viral immune proteins, including IFN-α, IFN-β, IFN-γ, CXCL1 (KC), TNF-α, CCL2 (MCP-1), IL-12p70, CCL5 (RANTES), IL-1β, CXCL10 (IP-10), GM-CSF, IL-10, and IL-6. The assay was validated for specificity, linearity, cross-reactivity, and inter and intra-assay precision. The assay was further validated using biological samples to quantify analyte concentration changes in response to poly (I:C) stimulation compared to untreated controls.
This multiplex panel is a robust tool for measuring the concentration of anti-viral inflammatory mediators in murine biological samples, and offers greater efficiency and broader dynamic ranges compared to conventional ELISA assays.
Hypoxia and monomeric IgE activate common signaling pathways leading to chemokine CCL2 production in mast cells
Authors: RAMIREZ-MORENO ITZEL GUADALUPE , Ibarra-Sánchez Alfredo , Gonzalez-Espinosa Claudia 
Published: January 2023

Abstract
Although presence of mast cells (MCs) to sites with low oxygen concentration (such as inflamed tissues with compromised blood flux and solid tumors) has been reported, the effects of hypoxia on the cytokine profile produced by MCs under those conditions has not been fully explored. We used the murine B16 melanoma model to quantify the presence of MCs in normoxic or hypoxic zones of the tumor and, bone marrow-derived mast cells (BMMCs) subjected to a 1% O2 atmosphere to characterize the production of specific cytokines and chemokines by this cell type in hypoxic conditions. Hypoxic areas of melanoma tumors were identified with hypoxyprobe and the number of MCs was determined by confocal analysis of tryptase positive cells. Using this methodology, important MCs presence in hypoxic zones of the tumor was detected. In vitro, BMMCs subjected to hypoxia were able to produce VEGF and the potent chemokine CCL2. Chemokine production in hypoxia was sensitive to the antioxidant Trolox and the IKK inhibitor BAY117085. CCL2 production was also induce by monomeric IgE by a mechanism also sensitive to Trolox and BAY117085. CCL2 production triggered by hypoxia or monomeric IgE was sensitive to calcium chelator BAPTA and the L-type calcium channel blocker Nifedipine. Our results show that MCs are importantly recruited to hypoxic sites of melanoma tumors and that hypoxia induces the production of the chemokine CCL2 by a mechanism that require the production of reactive oxygen species (ROS) and intracellular calcium rise through the activation of L-type calcium channels. Also, data indicate that monomeric IgE triggers a very similar response than hypoxia does to lead to the production of CCL2 in mast cells.
Prenatal smoke exposure exacerbates lung inflammatory response by modulating neutrophil infiltration to the lungs after RSV infection
Authors: Cheemarla Nagarjuna R. , Naidu Shan , Guerrero-Plata Maria Antonieta 
Published: January 2023

Abstract
Exposure to second hand tobacco smoke (SHS) is associated with an increased frequency and severity of viral respiratory tract infections. The mechanisms of recurrence and susceptibility to viral pathogens after passive smoke exposure are multifactorial and include alteration of the structural and immunologic host defenses. Maternal smoking during pregnancy is associated with a number of adverse health effects in infants, including higher susceptibility to respiratory syncytial virus (RSV) infection and associated bronchiolitis. Given the extensive impact on human health, further studies are needed that identify the mechanisms responsible for the increased severity of respiratory viral infections in individuals exposed to cigarette smoke. In this work, we used a well-established mouse model of in-utero smoke exposure. Offspring (5-day old) were infected intranasally with RSV and sacrificed at different times post infection. Sample analysis indicated that in-utero SHS exposure led to increased lung inflammation and higher viral lung titers, as shown by lung histopathology and plaque assay, respectively. In further understanding the mechanism of inflammation, cytokine profile revealed a significant increase in CXCL1 (KC), a potent neutrophil chemoattractant. Thus, we investigated the infiltration of neutrophils in lungs of SHS and air-exposed mice. FACS analysis revealed not only that RSV-infected neonates exposed to in-utero SHS had a significant increase in neutrophil influx into airways compared to air-exposed mice, but also that their neutrophils were more activated and expressed higher levels of CXCR2. In conclusion, our results help elucidate the mechanisms underlying harmful effect of smoking during pregnancy.
Zika, dengue and yellow fever viruses induce differential anti-viral immune responses in human monocytic and first trimester trophoblast cells
Authors: Luo Huanle , Winkelmann Evandro , Fernandez-Salas Ildefonso , Li Li , Mayer Sandra V. , Daniz-Lozano Rogelio , Sanchez-Casas Rosa Ma , Vasilakis Nikos , Tesh Robert , Barrett Alan D. , Weaver Scott C. , Wang Tian 
Published: January 2023

Abstract
Zika virus (ZIKV) is a mosquito-borne flavivirus associated with severe neonatal birth defects, but the causative mechanism is incompletely understood. ZIKV shares sequence homology and early clinical manifestations with yellow fever virus (YFV) and dengue virus (DENV) and are all transmitted in urban cycles by the same species of mosquitoes. However, YFV and DENV have been rarely reported to cause congenital diseases. Here, we compared infection with a contemporary ZIKV strain (FSS13025) to YFV17D and DENV-4 in human monocytic cells (THP-1) and first-trimester trophoblasts (HTR-8). Our results suggest that all three viruses have similar tropisms for both cells. Nevertheless, ZIKV induced strong type 1 IFN and inflammatory cytokine and chemokine production in monocytes and peripheral blood mononuclear cells. Furthermore, ZIKV infection in trophoblasts induced lower IFN and higher inflammatory immune responses. Placental inflammation is known to contribute to the risk of brain damage in preterm newborns. Inhibition of toll-like receptor (TLR)3 and TLR8 each abrogated the inflammatory cytokine responses in ZIKV-infected trophoblasts. Our findings identify a potential link between maternal immune activation and ZIKV-induced congenital diseases, and a potential therapeutic strategy that targets TLR-mediated inflammatory responses in the placenta.