bluuebunny/crossref_metadata_2025_split huggingface dataset:
Biophysical characterization of the adhesion between enveloped virus and phosphatidylserine-binding proteins
Authors: Zhang Xiaohui , Cao Wenpeng , Jagota Anand 
Published: January 2023

Abstract
Enveloped viruses are viruses that have a lipid membrane. Many enveloped viruses, such as the emerging virus Ebola, are pathogenic and of great clinical importance. Recent studies have shown that the outer lipid leaflets of viral envelopes contain phosphatidylserine (PS), which serves as an important attachment factor for enveloped virus entry into host cells. The envelope PS interacts with cellular PS receptors in a manner that mimics apoptotic body-PS receptor interactions during the engulfment of dead cell debris by phagocytes. The PS receptors utilized by enveloped viruses for viral internalization are designated phosphatidylserine-mediated virus entry enhancing receptors (PVEERs). While the importance of PVEERs in mediating viral entry has been demonstrated, the biophysical properties of the PVEER-virus interactions, including the detailed kinetics of PVEER-virion-associated PS binding, have not been comprehensively evaluated and compared. Using Ebola virus (EBOV) as the model system, this study investigated and compared the interactions between model Ebola viral particles and PVEER family proteins that are known to mediate enveloped virus attachment and entry. We used single-molecule force spectroscopy to quantify the specific interaction forces between PS or model EBOV viral particles and PVEERs. Based on the dynamic force spectra of various PS-receptor interactions, our results suggest that PS-PVEER interactions are mechanically comparable to antibody-antigen interactions, and that Gas6 and MFG-E8 may generate more force-resistant intersections than TIM-1 and ProteinS. The findings can help explain the biophysical mechanisms underlying the PVEER-mediated entry of enveloped virus into cells.
Evaluation of miRNAs role in the immunopathogenesis of Microcephaly caused by ZIKV in experimental models
Authors: Polonio Carolina Manganeli , Brandão Wesley Nogueira , Cugola Fernanda , Zanluqui Nagela , Oliveira Lilian , Garcia Marilia , Braga Patricia , Peron Jean Pierre S 
Published: January 2023

Abstract
Viral infections have always been the cause of serious human diseases, usually increasing rates of morbidity and mortality worldwide. In 2015, the flavivirus Zika virus (ZIKV) caused an alarming increase in the number of babies born with microcephaly. Genetic differences, mainly related to immune response, are known to influence susceptibility to infection, once is very important in blocking viral replication and may be modulated by several different factors, in which, miRNAs play a key role. Still very little is known about the involvement of miRNAs during ZIKV infection. In this context, we evaluated the role of miRNAs during ZIKV experimental infection. We performed miRNAs analysis in vitro using CNS cells and, in vivo, with SJL animals susceptible to infection. Analyzing human neurons, we observed that ZIKV was able to reduce miR-9 and this reduction is known to cause cerebral cortex thickness reduction and ventriculomegaly, characteristics similar to microcephaly. In human neuronal precursor cells, ZIKV was able to reduce miR-194, miR-302b and miR-302c expression and consequently increased MXD1, a transcriptional repressor. On the other hand, ZIKV infection increased miR-295 and miR-302d expression leading to a reduction of AHR repressor and molecules involved in neurodevelopment, neuroD4, and neuroD6. In SJL fetal brain a miR-146 increase leading a decrease of IRAK1, favoring viral replication. Taken together, these data are indicative that ZIKV is able to modulate miRNA profile to regulate important molecules involved in neurodevelopment and antiviral immune response, evidencing the importance of these regulatory molecules, which may help us to better understand immunopathogenic mechanisms of microcephaly caused by ZIKV
The divergent evolution of cytomegalovirus-encoded CD48 homologs contributes to the expansion of the immunevasin repertoire
Authors: Angulo Ana , Martínez-Vicente Pablo , Farré Domènec , Engel Pablo 
Published: January 2023

Abstract
The genesis of gene families by capture of host genes and their subsequent duplication is a crucial process in the evolution of large DNA viruses. CD48 is a GPI-linked cell surface protein containing two extracellular immunoglobulin (Ig) domains. Via its N-terminal Ig domain, CD48 interacts with the cell surface receptor 2B4, modulating leukocyte cytotoxicity. We previously reported the presence of five CD48 homologs (vCD48s) in two related cytomegaloviruses, derived from a common host CD48 ancestor gene acquired by retrotranscription. Recently, we examined one member of this family, A43, showing that it acts as a functional viral decoy receptor, impairing 2B4-mediated NK cell cytotoxicity. Here, we have characterized the rest of the vCD48s. We show that they are highly glycosylated type I transmembrane proteins which display remarkably distinct structures, biochemical properties, locations, and temporal kinetic classes. Molecular modeling of the N-terminal Ig domains of these vCD48s evidence significant changes in the number and length of their β-strands and inter-sheet loops that participate in the interaction of CD48 with 2B4, and accordingly we found that none of these molecules binds to 2B4. Interestingly, we determined that two of them, S30 and S31, have new targets in T or B lymphocytes. In particular, S30 tightly interacts with CD2, a T- and NK-cell adhesion and costimulatory molecule whose primary ligand is CD58. Thus, altogether our results show how a key host immune receptor captured by a virus can be subsequently remodeled during viral evolution to yield new molecules with shifted binding specificities to additional immune targets, expanding the repertoire of viral immunevasins.
Zika virus evolution in the presence of dengue virus-elicited cross-reactive immunity
Authors: Nava Jose Angel Regla , Wang Ying-Ting , Fontes-Garfias Camila R , Syed Thasneem , Gonzalez Andrew , Viramontes Karla , Valentine Kristen M , Terskikh Alexey V , Gleeson Joseph G , Shi Pei-Yong , Shresta Sujan 
Published: January 2023

Abstract
Zika virus (ZIKV), belongs to Flavivirus, causes neurological disorders including Guillain Barré syndrome and fetal microcephaly, and is responsible for widespread epidemics in countries where other flaviviruses, such as dengue (DENV), and yellow fever viruses, are endemic. However, how the cross-reactive, pre-existing immunity impacts the evolution of flaviviruses is poorly understood. To mimic the natural transmission cycle of ZIKV between vertebrate (human) hosts with pre-existing flavivirus immunity and invertebrate hosts (mosquito), ZIKV was subjected to 10 alternating cycles of passaging between DENV-immune mice and mosquito cells. We identified two mutations in the passaged ZIKV strains- one in the NS2B gene (I39V), both after passage in DENV-naïve and DENV-immune mice, and a mutation in the E gene (T470M), only after passage in DENV-immune mice. These mutations increase ZIKV virulence and pathogenesis in non-pregnant and pregnant Ifnar1−/− mice and abrogate cross-protection mediated by the pre-existing DENV immunity. In addition, ZIKV strain with the I39V mutation was more virulent than the parental ZIKV strain in human fetal neural progenitor cells (NPCs), and ZIKV variants in position 39 of the NS2B protein were observed in recent clinical isolates of ZIKV. Our data thus identify a mutational hotspot in the ZIKV NS2B protein and a mutation in the ZIKV E protein that arose as a consequence of the pre-existing immune pressure. Our results uncover the evolutionary strategies by which ZIKV can become pathogenic in both DENV-naïve and DENV-immune populations. Mechanistic studies of evolution in the presence and absence of cross-reactive immunity will benefit the development of safe and efficient flavivirus vaccines.
Features of Neuropsychiatric Lupus in Nba2 mouse model
Authors: Browne Kim , Zhang Emily , Sullivan James , Evonuk Kirsten Scarlett , Liu Jessica , DeSilva Tara M , Jorgensen Trine N 
Published: January 2023

Abstract

Background
Neuropsychiatric lupus (NPSLE) is a poorly understood manifestation of the autoimmune disease systemic lupus erythematosus (SLE). NPSLE is characterized by symptoms including anxiety and depression, and recent data have suggested a pathogenic role for interferon-alpha (IFNα). Additionally, IFNα can diminish the production of the neurotransmitter serotonin, suggesting a causative link between IFNα and NPSLE. B6.Nba2 mice develop SLE in an IFNα-dependent manner making it a valuable model for the study of NPSLE.

Methods
Age-matched female B6.Nba2 and B6 mice were tested. Behavioral studies included open field testing (OFT), elevated plus maze (EPM), and fear conditioning (FC) tests for detection of anxiety-like behavior, and forced swim test (FST) for detection of depression-like behavior. Serum serotonin and dopamine levels were measured by ELISA and serotonin receptors were enumerated by IHC.

Result
Results from both OFT (p=0.001), EPM (p&lt;0.05) and FC (p&lt;0.05) tests supported a strong anxiety phenotype in B6.Nba2 female mice, while data from FST (p&lt;0.05) suggested a depressive phenotype as well. B6.Nba2 females also showed significantly less serum serotonin (p&lt;0.0001) and dopamine (p&lt;0.05) levels than age-matched B6 females.

Conclusion
Our data suggests that the B6.Nba2 mouse model of SLE expresses an anxiety and depressive phenotype resembling NPSLE. Along with our previous data showing that IFNα levels are upregulated and required for disease development, our results suggest a positive correlation between IFNα, reduced serotonin, and an anxiety phenotype. Current studies are evaluating the effect of genetic IFNα-receptor manipulation in brain cells and the direct relationship between IFNα and NPSLE.
T-cells from NOD-PerIg mice transfer aggressive peripheral neuritis to NOD. scid recipients
Authors: Racine Jeremy J , Chapman Harold D , Doty Rosalinda , Cairns Brynn M , Hines Timothy J , Tadenev Abigail L.D. , Anderson Laura C , Green Torrian , Dyer Meaghan , White Jacqui , Ettinger Rachel , Burgess Robert W , Serreze David V 
Published: January 2023

Abstract
Recently, we created NOD mice transgenically expressing a peripherin reactive B cell receptor (BCR) inducing rapid onset T1D (NOD-PerIg). We unexpectedly discovered that in addition to T1D, NOD.scid recipients receiving transferred NOD-PerIg T-cells or whole splenocytes developed a visible hind leg weakness caused by aggressive limb neuritis. Neurobehavior phenotyping failed to uncover any measurable clinical metric that could predict onset of visible hind leg symptoms. Nerve conduction velocities were measured on hind limbs in mice that made it to the critical 15-weeks post transfer window, and revealed that most mice with clinical symptoms of neuritis had little to no discernible nerve conduction, while those without clinical symptoms of neuritis had near normal NOD.scid-like levels of nerve conduction. A longitudinal flow cytometry analysis of sciatic nerves at onset of diabetes, onset of visible hind leg symptoms, or at the end of incidence revealed that mice with clinical symptoms of neuritis had increased CD45+ cells amongst live cells compared to controls, and increased T-cells amongst CD45+ cells compared to both controls and mice without clinical symptoms. Depletion of either CD4 or CD8 T-cells after transfer of whole NOD-PerIg splenocytes led to the complete prevention of diabetes development. Interestingly, CD4 but not CD8 T-cell depletion led to the absence of any clinical manifestation of neuritis by 20 weeks post transfer. Finally, we have observed lymphocyte infiltration in both primary NOD-PerIg and NOD mice, likely explaining the occasional NOD mouse that has developed neuritis in The Jackson Laboratory’s extensive NOD collection.
IL-27-producing B cells synergize with IFNγ to promote IgG2a and IgG1 antibody responses
Authors: Yan Hui , Wang Rui , Wang Jingwei , Wu Shuai , Fernandez Maria , Rivera Carlos Eduardo , Moroney Justin B , Li Xiaodong , Zhang Nu , Zan Hong , Meng Xiangzhi , Zhang Fushun , Kedl Ross , Min Booki , Hunter Christopher A , Xiang Yan , Casali Paolo , Xu Zhenming 
Published: January 2023

Abstract
How B cells integrate innate receptor and adaptive T cell signals in the class-switched antibody response needs be better understood. By using cytokine gene knockout and reporter mice, in vitro B cell cultures and single-cell analyses, we have identified a subset of B cells that produces IL-27 as signature cytokine upon priming by TLR ligands, which up-regulate CD40 expression, and exposure to Tfh cell stimuli CD154 and IL-21. B cell IL-27 induction depends on NF-kB activated by linear ubiquitin assembly complexes and up-regulated Baft3 transcription factor, which are recruited to the Il27p28 locus, as made accessible by CD154 and IL-21 stimulation. IL-27-producing B cells are bystander B cells that are inefficient in secreting antibodies, but express high levels of adhesion molecules which engage other activated B cells. Together with IFNg, they boost proliferation, survival and class-switching to IgG2a of B cells stimulated with CD154 and IL-21. Accordingly, mice with B cell-specific ablation of IL-27 and IFNg receptors display severe defects in germinal center B cell and plasma cell differentiation, leading to abrogation of IgG2a and markedly reduced IgG1 responses. Thus, IL-27-producing B cells function as “helper” cells to promote differentiation of antigen-specific B cells for optimal IgG responses. As such, they play a critical role in the production of effective anti-viral antibodies and, possibly, modulation of anti-tumoral immunity.
Neutral sphingomyelinase 2 regulates inflammatory responses in monocytes/macrophages induced by TNF-α
Authors: Alrashed Fatema , Ahmad Zunair , Thomas Reeby , Alghaith Abdulwa , Melhem Motasem , Ahmad Rasheed 
Published: January 2023

Abstract
Obesity is associated with elevated levels of TNF-α and proinflammatory CD11c monocytes/macrophages. TNF-α mediated dysregulation in the plasticity of monocytes/macrophages is concomitant with pathogenesis of several inflammatory diseases, including metabolic syndrome, but the underlying mechanisms are incompletely understood. Since neutral sphingomyelinase 2 (nSMase2; product of the sphingomyelin phosphodiesterase 3 gene, SMPD3) is a key enzyme for ceramide production involved in inflammation, we investigated whether the nSMase2 contributed to the inflammatory changes in the monocytes/macrophages induced by TNF-α. In this study, we demonstrate that the disruption of nSMase2 activity in monocytes/macrophages either by chemical inhibitor GW4869 or small interfering RNA (siRNA) against SMPD3 results in defects in the TNF-α mediated expression of CD11c. Furthermore, blockage of nSMase in monocytes/macrophages inhibited the secretion of IL-1b and MCP-1. However, inhibition aSMase activity did not attenuate CD11c expression and secretion of IL-1b and MCP-1. Phosphorylation of JNK, p38 and NF-κB resulting from TNF-α stimulation was also attenuated by the inhibition of nSMase. Moreover, NF-kB/AP-1 activity was blocked by the inhibition of nSMase2. Our human data show that SMPD3 gene expression was not only found to be elevated in obese individuals but also positively corelate with TNF-α elevated expression. These findings indicate that nSMase acts, in a part, as a master switch in the TNF-α mediated inflammatory responses in monocytes/macrophages.
Elevated Interferon Alpha in the Serum of Laboratory Strains of Mice and Porcine Serum by ELISA
Authors: DeJesus Elfie , Schwartz Barbara , Kisiel Greggory , Clark William , Lavoie Thomas 
Published: January 2023

Abstract
We examined Interferon (IFN)-Alpha and -Beta levels in pooled sera and individual bleeds from C57/Bl, Balb/C and CD-1 mice. 20 Individuals from each strain from two independent colonies were evaluated in commercial assays. The IFN-Beta ELISA detected IFN in 1 of the 60 individuals while the IFN-Alpha assay which detects all 14 subtypes measured IFN in 13 of 60. 5 individuals exhibited greater than 4 pg/ml of IFN-Alpha detected. Specificity was demonstrated by knockdown of the signal using non-biotinylated detection antibody. 14 of 54 porcine sera samples from 4 independent sources were positive for IFN alpha by an ELISA under development, and similar knock-down experiments suggest specificity. This is in contrast to human sera/plasma samples where less than 3% of healthy donor samples have quantifiable IFN-Alpha or -Beta above 0.5 pg/ml. This study may raise questions about the regulation of the IFN system in different species and the potential of these differences to affect experimental results in a variety of models.
Adarza’s ZIVA multiplex protein detection platform utilizes a label-free technology to simultaneously monitor murine and human inflammation cytokines in a CAR-T model system
Authors: Schmuke Jon J , Mueller Ernie , Carter Alun , Cooper Matt , Kabakibi Ayman 
Published: January 2023

Abstract
We have used Arrayed Imaging Reflectometry (AIR™), a label-free protein microarray technology that relies on the target binding-induced perturbation of an antireflective coating on the surface of a silicon biosensor, to create a multiplex antibody array for the detection of human and murine serum cytokines and inflammatory markers. The biosensor is capable of providing quantitative protein concentrations, with limits of detection in the low picograms per mL. The flexibility of this system can also reduce the need to generate a series of sample dilutions in order to get results in a limited detection range by enabling the development of each assay with sensitivity expectations for each target on a chip. Provided in a standard 96-well microplate format, the label-free nature of the array enables flexible creation of standard or custom panels for development of model systems. Here, we demonstrate a custom 22 plex panel developed in collaboration with WUGEN Inc. to provide multi-species cytokine analysis in humanized CAR-T cell models for the evaluation of biologic product efficacy and safety. Samples from in vitro and in vivo models developed for the evaluation of CAR-T activity were analyzed and demonstrated a novel platform for concurrent cross-species cytokine analysis on a single array. Results are evaluated for sensitivity, precision, and accuracy with ZIVA assays comparable to traditional ELISAs. With array capabilities from low to hundreds of plex combined with a hands-free, automation-friendly workflow enabled by the ZIVA instrument, the technology has been designed for the ultimate user-experience.
Oral epithelial IL-22/STAT3 signaling licenses IL-17-mediated immunity to oral mucosal candidiasis
Authors: Aggor Felix E.Y. , Break Timothy , Trevejo-Nunez Giraldina , Whibley Natasha , Coleman Bianca M. , Bailey Rachel , Kaplan Daniel H. , Naglik Julian R. , Shan Wei , Shetty Amol C. , McCracken Carrie , Durum Scott K. , Biswas Partha S. , Bruno Vincent M. , Kolls Jay K. , Lionakis Michail S. , Gaffen Sarah L. 
Published: January 2023

Abstract
Oropharyngeal candidiasis (OPC) is an opportunistic infection of the oral mucosa caused by the commensal fungus Candida albicans. IL-17 and IL-22 both mediate antifungal immunity yet activate distinct downstream signaling pathways. While much is known about IL-17-dependent immunity in OPC, the activities of IL-22 are less well delineated. We show that induction of Il22 is independent of Dectin-1, CARD9 and aryl hydrocarbon receptor (AhR) and is driven by IL-23 and the C. albicans pore forming peptide candidalysin. Despite similar induction requirements and cellular sources, IL-22 and IL-17 function non-redundantly during OPC and exert opposing roles in neutrophil recruitment. The IL-22 and IL-17 receptors are required in anatomically distinct locations; loss of IL-22RA1 in the oral basal epithelial layer (BEL) but not the suprabasal epithelial layer (SEL) causes susceptibility to OPC, whereas IL-17RA is needed in the SEL. Our data reveal that IL-22 is a major activator of STAT3 in the BEL during OPC. Moreover, loss of STAT3 in the BEL but not the SEL renders mice susceptible to OPC. Transcriptional profiling of RNASeq data linked IL-22/STAT3 to oral epithelial cell proliferation and survival, but also, unexpectedly, to driving an IL-17 gene signature. We show that IL-22 acts on the BEL to replenish the IL-17RA-expressing SEL, thereby restoring the ability of the oral epithelium to respond to IL-17. Consequently, IL-22 signaling in BEL ‘licenses’ IL-17R signaling in the oral epithelium, revealing spatially distinct yet cooperative activities of IL-22 and IL-17 in oral candidiasis. This work also suggests that oral thrush in Jobs’ syndrome patients may be caused by STAT3 impairments in the oral epithelium, not just Th17 cells.
A FUNCTIONAL TL1A/DR3 SYSTEM IS CRUCIAL FOR OPTIMAL GENERATION OF PATHOGENIC TH9 CELLS IN MURINE CROHN’S DISEASE-LIKE ILEITIS
Authors: Butto Ludovica , Menghini Paola , Aladyshkina Natalia , Buela Kristine-Ann , De Salvo Carlo , Osme Abdullah , Pizarro Theresa , Cominelli Fabio 
Published: January 2023

Abstract
New evidence suggests the implication of IL-9-secreting TH9 cells in the pathogenesis of Crohn’s disease (CD). The role of death receptor 3 (DR3) in the differentiation and function of TH9 cells in CD is not fully explored. Thus, here, we investigated the role of a functional DR3 receptor in disease progression and activation of downstream signaling pathways relevant to TH9 cells. We found that lack of DR3 ameliorated ileitis in SAMP/YitFc (SAMP,WT) mice, reduced by 40% Baft3, PU.1, il-9 mRNAs and IL-9 protein, increased by 3-fold Id3 mRNA in ileal tissues (P≤0.003). TH9 cells from DR3−/−×SAMP (KO) mice harbored 2-fold less il-9 mRNA and IL-9 and TNF proteins (P≤0.0002), 5-fold higher IL-10 protein than WT TH9 cells (P&lt;0.0001). RNA-seq analysis of WT and KO TH9 cells is currently being analyzed. Rag2−/− recipients of KO TH9 cells developed less severe colitis (P=0.005). Histology score of ilea and colons from SAMP×Rag2−/− is currently being evaluated. The frequency of KO TH9 cells producing IL-9 in Rag2−/− MLNs was 2-fold lower than that of mice reconstituted with WT TH9 cells (P=0.02). The frequency of IL-9-producing KO TH9 cells in SAMP×Rag2−/− SPLs and MLNs was 2-fold lower than that of mice reconstituted with WT TH9 cells (P≤0.006). In conclusion, DR3 receptor heavily contributes to IL-9-signaling pathway activation and it is required for the secretion of higher amount of IL-9 protein by TH9 cells in SAMP mice. KO TH9 cells were less colitogenic that those isolated from WT mice, indicating a crucial role of a functional DR3 receptor in TH9 pathogenicity. Collectively, our results hold great translational significance by showing that modulation of DR3 signaling may be a novel therapeutic target for the treatment of CD.