bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of a Multiplex Bead Serological Assay for Use in Global Laboratories to Assess Population Immunity to Measles and Rubella
Authors: Coughlin Melissa M , Matson Zachary , Sowers Sun B , Priest Jeffery , Smits Gaby , van der Klis Fiona , Hickman Carole , Scobie Heather , Rota Paul , Bankamp Bettina 
Published: January 2023

Abstract
Serosurveys are an important tool for the evaluation of population immunity and guidance of immunization activities. The multiplex bead assay (MBA) offers multiple benefits over standard serological assays and this study sought to compare the use of MBA to measure humoral immunity to measles and rubella compared to the enzyme immunoassay (EIA) for both viruses and the plaque reduction neutralization (PRN) for measles. Multiplex beads coupled with laboratory-produced purified measles whole virus antigen (WVA) correlated better to EIA and PRN than baculovirus-expressed measles nucleoprotein (N); therefore, a commercial antigen source of measles WVA was evaluated. The commercial measles WVA demonstrated significant correlation using Pearson’s correlation coefficient to the laboratory-produced WVA MBA (R=0.995, p=&lt;0.0001) and demonstrated a strong linear relationship between the two WVA MBA assays (R2= 0.926). The commercial measles WVA MBA correlated strongly to PRN (R=0.9530, p=&lt;0.0001) with a linear relationship comparable to the laboratory-produced WVA and PRN (R2=0.716 and R2=0.768, respectively). Receiver operator curve (ROC) analysis resulted in a seroprotection cutoff of 153 mIU/ml, similar to the established correlate of protection of 120 mIU/ml, with a sensitivity of 98% and a specificity of 84%. Comparison of the rubella MBA to EIA demonstrated a strong linear correlation of the two assays (R=0.959 and R2=0.919). ROC analysis yielded a cutoff of 9.36 IU/ml, similar to the accepted cutoff of 10 IU/ml for seroprotection with a sensitivity of 99% and a specificity of 100%. The results presented here support the use of the MBA as a tool to conduct multiplexed serosurveys to assess population immunity to measles and rubella.
Impact of pre-existing specific influenza antibodies as a determinant of disease severity on influenza virus infected patients: a multicenter observational cohort study
Authors: Aydillo GOmez Teresa A , Merino Alba Escalera , Aslam Sadaf , Carratala Jordi , Krammer Florian , Garcia-Sastre Adolfo , Matia Elisa Cordero 
Published: January 2023

Abstract
It is unclear to what extent pre-existing antibody-mediated immunity shapes influenza virus infection in humans. Similarly, the influence of previous history of influenza vaccination on disease severity and clinical outcome is not well understood. In here, we aimed to determine the balance of pre-existing antibodies against the hemagglutinin (HA) and neuraminidase (NA) protein of influenza virus that correlates with protection in humans. We measured hemagglutination inhibition (HI) titers and anti-HA (stalk and full-length) and anti-NA antibodies at the onset of the influenza infection on a cohort of H1N1/Cal09 influenza virus-infected patients (solid organ transplant recipients, SOTRs). Results demonstrated that the antecedent of TIV vaccination was the only controllable factor associated with lower risk of severity of disease in influenza virus-infected SOTRs. The presence of lower respiratory symptoms (LRS) significantly determined disease outcome at hospital admission (severe vs. mild, 9, 75% vs. 19, 32·1%, OR 95% 6·3, p=0·01). Moreover, a predicted probability curve for both anti-HA full-length and stalk (OR 0·13, p=0·02 and OR 0·06, p=0·03, respectively) demonstrated that the probability of developing LRS decreased with increasing assay log-values. However, multivariate logit model adjusted for HI, anti-HA (full-length and stalk) and anti-NA titers demonstrated that only anti-HA stalk antibodies were independently associated with influenza severity. Our study demonstrated that when HI antibodies fail to confer protection from symptomatic influenza infection, pre-existing HA stalk antibodies protected against severe disease in SOTRs.
Structural basis of neutralization of Powassan virus by monoclonal antibodies
Authors: Errico John M , VanBlargan Laura A. , Nelson Christopher A. , Diamond Michael S. , Fremont Daved H. 
Published: January 2023

Abstract
Powassan virus (POWV) is an emerging tick-borne flavivirus endemic to the northern and northeastern United States. Severe human POWV infection is characterized by fatal encephalitis in 10% of patients and long-term neurological sequelae in 50% of survivors. Little is known about humoral immunity to POWV, and there are currently no licensed vaccines or therapeutics available for treatment. We have generated a panel of 23 murine antibodies against POWV that separate into six epitope groups based on competitive binding. Two of the groups target domain III of the viral envelope protein; mapping by alanine scanning mutagenesis places their epitopes on the A-strand and C-C′ loop of domain III, respectively. These DIII-specific mAbs display potent neutralization in vitro and robust protection in a mouse model of POWV infection. X-ray crystallographic analysis reveals that POWV-80, a C-C′ loop antibody, adopts a binding orientation similar to a class of potently neutralizing flavivirus antibodies that target the lateral ridge epitope on domain III, while POWV-61 binds primarily to the A-strand and B-C loop. Our results indicate that antibodies to POWV can target epitope regions similar to those observed on mosquito-transmitted flaviviruses by employing unique binding modes that may facilitate enhanced neutralizing and protective efficacy. Current efforts are focused on defining the determinants of antibody potency and understanding antibody binding in the context of the virion.
Severe fever with thrombocytopenia syndrome (SFTS) induce innate immunity disorder particularly in monocyte subsets
Authors: Zhang Li , Fu Yuxuan , Wu Allen 
Published: January 2023

Abstract
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever with a high mortality rate in humans, which is caused by SFTS virus (SFTSV), a novel phlebovirus in the Bunyaviridae family, is tick borne and is endemic in Eastern Asia. Previous study found that SFTSV can infect and replicate in macrophages in vivo and in vitro. However, the role of macrophages in virus replication and the potential pathogenic mechanisms of SFTSV in macrophage remain unclear. In this study, we provided evidence that the SFTSV infection drove macrophage differentiation skewed to M2 phenotype, facilitated virus shedding and resulted in viral spread. We showed evidence that miR-146a and b were significantly upregulated in macrophages during the SFTSV infection, driving the differentiation of macrophages into M2 cells by targeting STAT1. Further analysis revealed that the elevated miR-146b but not miR-146a was responsible for IL-10 stimulation. We also found that SFTSV increased endogenous miR-146b-induced differentiation of macrophages into M2 cells mediated by viral non-structural protein (NSs). The M2 skewed differentiation of macrophages may have important implication to the pathogenesis of SFTS.
Myeloid cell populations are influenced in patients’ peripheral blood during acute SFTS virus infection. SFTSV specifically targets human CD14+ blood monocytes, inducing phenotypic shift. Our studies imply that monocytes/macrophages may support persistent SFTSV viral infection. However, the evidence that monocytes/macrophages are directly involved in the development of SFTS in patients, particularly those in the acute phase.
HIV-infected apoptotic hepatocytes program macrophages for inflammasome development: role of alcohol
Authors: Osna Natalia A , New-Aaron Moses , Ganesan Murali , Dagur Ragubendra S , Poluektova Larisa Y 
Published: January 2023

Abstract
Liver injury is a second cause of non-AIDS mortality in HIV-infection, and the progression to end-stage liver disease is more rapid in alcohol abusers. Hepatocytes comprise 80% of liver cells and are strong ethanol metabolizers. To elucidate whether hepatocytes are HIV-infected, we measured the kinetics of HIVgag RNA (RT-PCR) and p24 (Western blot) expression in Huh7.5-CYP (hepatocyte-like cells) exposed to HIV-1ADA in the presence or absence of acetaldehyde-generation system (AGS). The measurements were done at days 1, 3 and 5 post-infection. We found that the highest levels of HIV markers were observed at day 1, with further decrease by day 3 and 5. AGS treatment substantially increased these levels. The accumulation of HIV in hepatocytes was accompanied by activation of oxidative stress, judged by increasing reactive oxygen species (ROS) production at day 3 and 5 post-infection and enhanced expression of 4-hydroxynonenal (4HNE) protein adducts, which increased caspase 3 cleavage. To mimic proapoptotic effect of acetaldehyde and study the involvement of apoptotic hepatocytes in liver inflammation, we incubated apoptotic HIV-infected or uninfected Huh7.5-CYP cells with monocyte-derived human macrophages (MDMs) and then measured inflammasome activation based on NLRP3, caspase 1, IL-1β and IL-18 mRNAs. Activation of inflammasome in MDMs was more prominent when HIV-infected apoptotic cells were engulfed by MDM compared with uninfected cell engulfment. We conclude that acetaldehyde synergies with HIV to induce oxidative stress and apoptosis in hepatocytes. The engulfment of apoptotic HIV+ hepatocytes leads to inflammasome activation in macrophages, thereby promoting progression to liver injury.
Glutamine metabolism is required for alveolar macrophage proliferation
Authors: Wang Min , Zhu Bibo , Sun Jie 
Published: January 2023

Abstract
Alveolar macrophages (AMs) are the most abundant immune cells in the lung during homeostasis. AMs are critical for normal lung development, surfactant metabolism and host defense against various respiratory pathogens. Despite being terminally differentiated cells, AMs are able to proliferate and self-renew to maintain their compartment without the input of the hematopoietic system in the adulthood during homeostasis. However, the molecular and metabolic mechanisms modulating AM proliferative responses are still incompletely understood. Here we have investigated the metabolic regulation of AM proliferation and self-renewal. We found that mitochondrial metabolism is critical for AM proliferation in vitro and in vivo. Interestingly, inhibition of glucose uptake or fatty acid oxidation did not significantly impact AM proliferation. Rather, inhibition of the glutamine metabolic process abolished AM proliferation in vitro and in vivo following their partial depletion during influenza infection. We further demonstrated that glutamine metabolism was required for AM mitochondrial respiration. Our data indicate that glutamine is a critical substrate fueling mitochondrial metabolism required for AM proliferation. Our study is expected to shed light on the metabolic regulation of AM maintenance and repopulation during homeostasis and following acute respiratory viral infection.
New insights into the role of N-glycosylation involved in molecular immune checkpoint regulation by human cytomegalovirus glycoproteins with the implication in cancer therapy
Authors: Nemcovicova Ivana , Benko Mario , Lenhartova Simona , Kempova Viera , Nemcovic Marek 
Published: January 2023

Abstract
The regulation of immune checkpoint molecules, such as BTLA or TIGIT, on immune cells occurs during persistent viral infections. These pathways are important for preventing immune-driven pathology but can also limit immune-mediated clearance of the infection. The recent success of immune checkpoint blockade in cancer therapy suggests that targeting these pathways would also be effective for preventing and treating a range of infectious diseases. Here, we report our current study of immune checkpoint pathways in the pathogenesis of human cytomegalovirus and discuss the potential for therapeutically targeting these pathways in this setting.
Human cytomegalovirus (HCMV) belongs to the beta herpes virus family that has co-evolved with the host immune system. UL144 and UL141 are found exclusively in clinical HCMV strains; while UL144 encodes a structural homologue of the herpesvirus entry mediator HVEM (target of BTLA), UL141 mimics TIGIT immunoreceptor. UL144 plays a role in virus-mediated immune evasion by transmitting inhibitory signals to downregulate T-cell responses and UL141 play significant role in NK cell evasion mechanism. Their sequences show considerable number of N-linked glycosylation sites located mostly in extracellular part. Many of them are not present in other viral species that suggest this glycosylation is important in human setting and may play a role in ligand-binding recognition. Here, we present characterization of recombinant HCMV glycoproteins by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and intact protein analysis. The individual glycan profiles followed protein-binding studies showed specific function of glycosylation in immune recognition.
Role of Tonsillar Innate and Adaptive immune cells during SIV infection in Rhesus macaques
Authors: Liyanage Namal , Shukla Rajni Kant , Reinhold-Larsson Nicole , Duncan Michael , Hatharasinghe Amila 
Published: January 2023

Abstract
In recent years, the incidence of HIV has decreased, however it continues to spread globally. 70-90% of individuals infected with HIV develop oral mucosal infections. Interestingly, HIV infected patients are at higher risk of developing Epstein–Barr virus (EBV)-associated B cell malignancies in tonsils. However, the potential role of oral immunity in pathogenesis of oral lesions is unknown. Tonsils are considered oral-pharyngeal mucosal associated lymphoid tissues and play an important role in oral mucosal immunity. However, detailed study of human tonsillar immune cells during HIV infection is restricted due to limited sample availability. In this study, we investigated the role of innate and adaptive immune cells in macaque tonsils by high dimensional flow cytometry analysis. Our data revealed a novel innate B cell subset that express NK cell receptors (NKB) significantly increased during SIV chronic infection in tonsils. We also found loss of ploy functional memory CD8 T cells during SIV infections. Tonsillar IL17 producing Cytotoxic T cells (Tc17+) were significantly increased during infection despite a loss in Th17 cells. Consistent with our previous studies in gut mucosal tissues, we found a significant loss of NCR+ILC3 in the tonsils during SIV infection. However, in contrast to the gut, we observed an expansion of oral CD56+ NK cells in tonsils during chronic SIV infection. These data suggest that the SIV associated immune responses are significantly distinct in the tonsils compared to other mucosal tissues. These data extend our understanding of the oral innate immune system during SIV infection and could aid future studies in evaluating the role of tonsillar immune cells during HIV associated oral mucosal infections.
Oligoadenylate Synthetase 1 enhances DNA sensor cGAS translation to mediate WNV antiviral activity
Authors: Perez Joseph , Harioudh Munesh K. , So Lomon , Nair Sharmila , McCormick Kevin , Ghosh Arundhati , Shao Lulu , Srivastava Rashmi , Ebert Thomas , Atianand Maninjay , Hornung Veit , Diamond Michael S. , Savan Ram , Sarkar Saumendra N. 
Published: January 2023

Abstract
Interferons inhibit virus replication through the expression of interferon stimulated genes (ISGs). We have found that a specific isoform of one such ISG, Oligoadenylate Synthetase 1 (OAS1) limits host susceptibility to West Nile Virus (WNV) infection through a non-canonical mechanism. This OAS1 isoform (OAS1 P46) in humans is generated due to an alternative splice acceptor site at the C-terminus of OAS1 gene. The SNP rs10774671 at this site has been associated with disease severity to WNV. We show that human OAS1-KO cells have lower basal levels of cGAS protein and can be rescued by OAS1 P46 independent of its enzyme activity. Additionally, through RNA-seq, SILAC, polysome profiling and radiolabeling experiments, we show that OAS1 does not regulate mRNA transcription but instead enhances protein translation of a select set of mRNAs, thereby increasing the steady state and induced levels of specific proteins with antiviral properties. Inducible expression of OAS1 P46 in cGAS-KO cells does not suppress WNV replication, suggesting that the antiviral activity of OAS1 is mediated through cGAS. We also have established functional equivalence between OAS1 P46 and a mouse ortholog, Oas1b (no enzyme activity), which similarly affects WNV susceptibility. Oas1b inhibits WNV infection and pathogenesis in vivo and inhibits WNV infection in vitro in cGAS-dependent manner. Through RNA-protein crosslinking experiments we have identified target mRNAs that bind to OAS1 and have demonstrated increased sensitivity of WNV in Oas1b RNA binding mutants. Our findings suggest a novel mechanism of OAS1 in which it binds to target mRNAs, enhances the translation of these RNAs and limits virus infection.
Characterization of a novel dengue virus strain and its implication for vaccine development
Authors: Chen Rita E , Errico John M , Gordon David , Dowd Kimberly A. , Cardosa M. Jane , Sariol Carlos A. , Kallas Esper G. , Sekaly Rafick-Pierre , Vasilakis Nikos , Pierson Ted , Fremont Daved H. , Diamond Michael S. 
Published: January 2023

Abstract
Dengue virus (DENV) causes up to 390 million infections yearly worldwide. Currently, four serotypes, DENV 1–4, circulate between Aedes mosquitoes and humans and can cause severe dengue, which has been associated with secondary DENV infection by a different serotype. DENV strain DKE-121 was recently isolated from Malaysia and is 12–38% different in its envelope protein from DENV 1–4. While previous studies of DENV genetic variation have described up to 3% amino acid divergence within a serotype, this virus differs by up to 12% from DENV4, suggesting its possible classification as a new serotype. The potential of a new DENV serotype emerging into circulation raises concerns of increased risk of severe dengue and uncertainty of how protective current tetravalent vaccine efforts would be. We tested the ability of serum from mice, non-human primates (NHPs), and humans that were infected or vaccinated with DENV4 and DKE-121 to neutralize infection of DENV4 and DKE-121. NHPs and humans immunized with DENV4 had 2 and 5-fold higher neutralizing titers (EC50 values), respectively, against the homologous DENV4 than DKE-121. However, mice boosted with DENV4 had similar neutralizing titers against DENV4 and DKE-121. In comparison, DKE-121 infection in NHPs and mice elicited 3-fold and up to 21-fold, respectively, higher titers against DKE-121 than DENV4. In addition, DKE-121 was poorly neutralized by type-specific anti-DENV4 monoclonal antibodies (mAbs), and reciprocally neutralizing mAbs against DKE-121 did not inhibit DENV4 infection. Using polyclonal sera and mAbs, DKE-121 and DENV4 show substantive differences in antigenicity. Ongoing studies are aimed at determining how these differences affect antibody-mediated protection in vivo.
Immune responses in Mayaro infected mice and its relation with viral replication and muscle damage
Authors: Neris Romulo , Figueiredo Camila , Leopoldino Daniel , da Silva Mariana , Almeida Juliana , dos-Santos Julio , Figueiredo Claudia , Bellio Maria , Bozza Marcelo , Miranda Iranaia 
Published: January 2023

Abstract
Mayaro virus (MAYV) is a tropical arbovirus first described in forest regions of the South America, and recently associated with urban circulation. Mayaro infections leads to a disease with high rate of persistent arthralgia and myalgia in humans. Despite this, pathogenesis and host immune response of MAYV infections are still limited. The aim of this work was to evaluate the immune response and cell profile during MAYV infections. We developed a disease model infecting SV129 wild-type (WT), SVA129 Type I Interferon receptor deficient (IFNAR−/−), C57BL/6 WT and C57BL/6 recombination activation gene-1 deficient (RAG−/−) mice, and accessing replication, tissue damage, and inflammation through time. While adult WT mice are resistant to infection, IFNAR−/− mice exhibited increasing viral load up to 6 days after infection, footpad swelling, lethargy and weight loss. Histological analysis showed extensive necrosis sites in infected IFNAR−/− mice and also presence of mononucleated cell infiltrate. We detected elevated expression of inflammatory cytokines like TNF, IL-6, KC, IL-1, MCP-1 and RANTES, and decreased TGF-beta in infected muscle. In RAG−/− mice, MAYV infection lead to a persistent replication being detected at blood and tissues up 40 days post infection. However, infected RAG−/− mice did not develop any signs of muscle damage or infiltrate during early and late infection and neither the expression inflammatory cytokines. So far, our data demonstrates that innate and adaptive immunity are critical to restrict MAYV replication. Adaptive immunity is also involved in MAYV-induced tissue damage. These results contribute in underlying MAYV pathogenesis and further experiments must to be made to elucidate the mechanisms involved.
Gp120 plays a role in the hypo-functionality of the cholinergic anti-inflammatory pathway in HIV infection
Authors: Samarani Suzanne , Aldabah Zainab , Ahmad Ali 
Published: January 2023

Abstract
We have investigated the functionality of the cholinergic anti-inflammatory pathway (CAP) in HIV-1 infected individuals. The pathway plays an important role in regulating inflammatory process in the body. Acetylcholine (ACh), a neurotransmitter, is the main effector molecule of the CAP. It is produced by CD4+ T cells, intestinal epithelial cells as well as by neurons in the body, and exerts its effects via nicotinic (n) and muscarinic (m) Acetylcholine receptors (AChR). Through these receptors, ACh modulates differentiation, proliferation and effector functions of immune cells in the body. Little is known about the regulation of the non-neuronal CAP in HIV infections. To investigate it, we measured concentrations of ACh and the Secreted Ly6/Urokinase type plasminogen activator Receptor-related Peptide-1 (SLURP-1, an allosteric ligand for the α-7nAChR) in the circulation of HIV-infected individuals. The concentrations of both ACh and SLURP-1 were increased in treatment-naïve HIV-infected individuals. In contrast to an increase in the concentrations of ACh and SLURP-1, the expression of the α-7nAChR was significantly decreased on immune cells in the virus-infected individuals. We investigated the effects of gp-120 on the expression of this receptor. We found that the treatment of the human PBMC with recombinant gp120 reduces the expression of α-7nAChR on immune cells. Interestingly, the treatment increases the binding of α-bungarotoxin, a ligand for nAChR. Collectively, these results suggest that the non-neuronal CAP is functionally compromised in HIV-infected individuals and gp120 plays a role in the hypo-functionality by down-regulating the expression of α-7nAChR on the peripheral blood immune cells.