bluuebunny/crossref_metadata_2025_split huggingface dataset:
Vitamin A deficient mice are highly susceptible to Streptococcus pneumoniae and exhibit a poor response to vaccination with Prevnar-13
Authors: Penkert Rhiannon , Hrincius Eike , Smith Amanda P , Smith Amber M , Rosch Jason , Hurwitz Julia L 
Published: January 2023

Abstract
Streptococcus pneumoniae (pneumococcus) is frequently found in the human respiratory tract and can cause a wide range of diseases from mild otitis to fatal pneumonia or meningitis. It is estimated that pneumococcus kills 1.6 million people each year, approximately half of whom are children in developing nations. Given that vitamin A deficiency (VAD) occurs in both developed and developing countries, we sought to explore the relationship between VAD and pneumococcal disease. Previously, we have shown that mice with VAD fail to clear respiratory virus infections efficiently, resulting in a prolonged state of inflammation. Here we show that mice with VAD are more likely to develop a severe lung infection following intranasal infection with pneumococcus compared to vitamin A sufficient controls. We also found that VAD increases morbidity and mortality from secondary pneumococcal infections in influenza-infected mice. This increased sensitivity to pneumococcal infections led us to investigate the effectiveness of the human pneumococcal vaccine Prevnar-13 (PCV) in the context of VAD. Vaccinated mice with VAD mounted only minimal IgM and IgG antibody responses toward PCV compared to controls. Upon challenge with an invasive strain (TIGR4) of pneumococcus, vaccinated control animals were nearly 100% protected from disease with no detectable bacteremia at 24 hours post-infection, whereas nearly 1/3 of vaccinated VAD animals developed bacteremia within 24 hours and ultimately succumbed to the infection. Our results suggest that clinical analyses of children in both developed and developing countries should be conducted to ensure that PCV successfully protects against pneumococcal disease in individuals with VAD.
A novel therapeutic vaccination delays cART-resistant HIV-1 reservoir rebound in HIV-1 infected humanized mice
Authors: Cheng Liang , Li Guangming , Li Dan , Zhang Zheng , Li Feng , Zurawski Sandra , Zurawski Gerard , Levy Yves , Su Lishan 
Published: January 2023

Abstract
Highly active anti-retroviral therapy (HAART) prevents disease progression but does not eradiate HIV, and this therapy often does not fully restore HIV patients’ immune status. Thus, there remains an unmet need to develop new stratagies to “cure” (eradication or control of HIV-1 with no ART) individuals of HIV infection. Modulating host’s cellular immune response is a promising way to cure HIV infection. We have developed a prototype vaccine with a string of 5 highly conserved T cell epitope rich regions of HIV-1 Gag, Nef and Pol fused to a monoclonal antibody that bind the antigen presenting cell activating receptor CD40(aCD40.HIV5pep). Here we use the humanized mouse model to evaluate the therapeutic effect of the aCD40.HIV5pep vaccine to chronic HIV infection. We first screened a batch of novel TLR adjuvants which have the potential to enhance the immunogenicity of aCD40.HIV5pep in vivo and showed that when vaccinated with Poly I:C as adjuvants, aCD40.HIV5pep can induce human CD8+ CTL response in humanized mice. Interestingly, we found that Poly I:C treatment can activate cART-resistant HIV-1 reservoir in in the presence of cART in chronically infected humanized mice. Finally, we showed that Poly I:C plus aCD40.HIV5pep therapeutic vaccination significantly delayed HIV virus rebound in HIV-1 infected humanized mice after withdraw of cART treatment. Our proof-of-concept study in humanized mice has significant implication for the development of therapeutic vaccine to cue HIV infection.
IGHV and IGHD encoding antibodies with exceptionally long CDR3H are predominant in the bovine neonatal B cells
Authors: Kaushik Azad Kumar , Pasman Yfke , Merico Daniel 
Published: January 2023

Abstract
We sequentially analyzed 18 RNA-seq libraries from three newborn calves as compared to their dams to understand the development of neonatal humoral immunity. A significant shift in neonatal transcriptome occurs within first week of life post-birth. A set of 110 differentially expressed immune-related genes is noted at birth that include complement, MHCII, chemokine receptors, defensins and cytokines etc. As compared to adults, IGKC is expressed at higher levels in the neonates, despite predominant IGLC expression at birth. The IGHM and IGHD transcripts are expressed at birth and IGHM achieves adult levels by day 7. This is followed by IGHA and IGHG transcripts 14–28 days post birth. Interestingly, most 3′ IGHV1S1(BF4E9) gene, known to encode antibodies with exceptionally long CDR3H, is preferentially expressed in neonatal B cells. In parallel, single longest IGHD2(DH2) gene that encodes such an atypical CDR3H is expressed at high levels at birth. It seems that IGHV1S1(BF4E9) gene has an important functional role in the development and expansion of bovine neonatal B-cell repertoire. To conclude, the bovine neonate acquires humoral immunocompetence by four weeks of neonatal life. [Supported by NSERC Canada Discovery Grant]
Indelible labeling and tracking of influenza virus infected antigen presenting cells
Authors: Langlois Ryan A , Fiege Jessica K 
Published: January 2023

Abstract
Influenza A virus is a seasonal pathogen with the potential to unpredictably cause catastrophic pandemics. The virus primarily replicates in epithelial cells of the upper and lower respiratory tract but can also infect a spectrum of other cell lineages, including cells of the immune system. Classically infected cells have been tracked through detection of virus products (RNA or protein) or through virus-derived reporters. However, these methods cannot define any potentially remaining infected cells after active replication has ceased. Using a novel virus expressing Cre recombinase to indelibly label infected cells in reporter mice, we unexpectedly found that epithelial cells were capable of surviving influenza virus infection long-term (Heaton and Langlois et al J. Ex. Med. 2014). We have subsequently discovered that infected CD45+ immune cells are also capable of surviving acute influenza virus infection in vivo. Importantly this virus-reporter system is able to distinguish between immune cells that are directly infected versus those that acquire virus protein/antigen exogenously. We find that both dendritic cells and macrophages, but not B cells, are directly infected within the lung and that the number of these cells increases during the acute infection phase. However, infected dendritic cells and macrophages do not persist in the lungs nor do they migrate systemically. Together these data demonstrate a new tool to track infected cells and defines the migration, turnover, and survival of infected antigen presenting cells.
Influenza A virus specific and Epstein-Barr Virus cross-reactive CD8 T cells during acute symptomatic flu infection.
Authors: Mishra Rabinarayan , Gil Anna , Aslan Nuray , Luzuriaga Katherine , Selin Liisa K 
Published: January 2023

Abstract
Previously we have shown expansion of cross-reactive CD8 T cells between Epstein-Barr Virus (EBV) and Influenza A virus in HLA-A2+ patients during infectious mononucleosis. During acute flu infection expanded IAV-M1-GIL58–66 specific CD8 T cells and IAV-M1-GIL58–66/EBV-BRLF-1-YVL109–117 and IAV-M1-GIL58–66/EBV-BMLF1280–288-GLC double-tetramer+ CD8 T cells were detected directly ex-vivo in 5 HLA-A2+ patients. IAV-M158–66, EBV-BRLF1119–117 and -BMLF1280–288 TCR repertoires changed over the course of infection and in comparison to healthy donors. After three weeks culture with Flu-M1 and EBV-BMLF1 ad EBV-BRLF1 peptides, CD8 T cells were sorted and analyzed by gene array in order to assess large-scale changes in immune responses in cognate or cross-reactive T cell populations in different patients. Global expression profile indicates cognate M1-GIL58–66 and BRLF1-YVL109–117 specific cells had similar immune-response gene signatures, but the EBV-BMLF1280–288 -GLC specific CD8 cells were more similar to the crossreactive populations. Crossreactive M1-GIL58–66/BRLF1-YVL109–117 cells from the BRLF1-YVL line were different in their activation status than the cognate BRLF1-specific cells, consistent with BRLF1-YVL ligand stimulation of different gene activation profiles in these two populations. These results indicate that during symptomatic flu infection there is an expansion of IAV/EBV crossreactive memory CD8 T cell responses. Ongoing studies are investigating whether EBV-IAV cross-reactive CD8+ T cells may contribute to immunopathology during acute IAV infection (NIH/NIAID PO1 AI 049320).
Identification of the immunological parameters that associate with severe influenza disease or recovery in mice
Authors: Forero Adriana , Nishida Andrew , Steuerman Yael , Gat-Viks Irit , Katze Michael G. , Morrison Juliet 
Published: January 2023

Abstract
The severity of influenza virus infection is largely determined by the interplay between the virus and the host. Understanding the pathological drivers of influenza A viruses serves to rapidly characterize the host response to newly emerging influenza viruses and derive prognostic signatures of disease severity. We implemented a differential network approach to uncover novel distinctions between the global transcriptional host response to highly pathogenic and low pathogenic H1N1, H7N9, H7N7, and H5N1 influenza A infections in BALB/c mice. We identified crucial mediators of inflammatory, coagulation, and tissue repair responses that discriminate between disease outcomes. It has been demonstrated that these responses are largely impacted by the immune cell influx into the site of infection. We examined the contribution of resident and lung-infiltrating immune cell quantity and activation state to disease phenotypes by implementing digital cell quantification (DCQ), which allowed us to model the in vivo dynamics of immune cells across viral strains based on time and strain-dependent infected lung transcriptional profiles. Through linear regression modeling, we revealed that distinct monocyte, granulocyte, DC, and T cell populations predict disease morbidity. Additionally, we demonstrated that both the kinetics of CD8+ T cell infiltration and the extent of their activation state, serve as a key predictor of immunopathological consequences of influenza infection. Overall, we provide a multidimensional analysis that delineates how innate immune responses relate to aberrant tissue repair responses and drive subsequent host adaptive immune response that control the severity of influenza virus disease in the murine model.
Prior administration of staphylococcal enterotoxin B augments the CD8+ T cell response to influenza A virus.
Authors: Meilleur Courtney Erin , Memarnejadian Arash , McCormick John K. , Haeryfar S.M. Mansour 
Published: January 2023

Abstract
Staphylococcal enterotoxin B (SEB) is a bacterial protein capable of binding up to 20% of the entire T cell repertoire. T cells that have come into contact with SEB initially proliferate and secrete inflammatory cytokines; however, this is followed by anergy or death of the affected cells. This population of deleted or functionally inactivated cells is likely to include T cell clones that are important for immunity to other pathogens such as influenza A virus (IAV). For this reason, we hypothesize that SEB exposure will alter the magnitude and breadth of the CD8+ T cell response to IAV. Using mouse model of intraperitoneal vaccination with IAV, we found that prior exposure to SEB actually increased the number of IAV-specific CD8+ T cells for several IAV epitopes during the primary phase of the immune response. The number of cells specific for these epitopes remained elevated even after the peak of the primary CD8+ T cell response had passed. In addition, repetition of the first experiment with several different naturally circulating and lab-adapted strains of IAV gave similar results, showing that the effect of SEB on virus-specific CD8+ T cells is the same for a potentially diverse range of IAV strains. Furthermore, the ability of the CD8+ T cells to kill target cells pulsed with cognate IAV antigen was also enhanced after SEB exposure, for both primary and memory CD8+ T cells. Curiously, we did not observe an effect of SEB when it is given in the context of a replicative IAV infection in the lungs. This work reveals an unexpected role for SEB as an enhancer of CD8+ T cell immunity in this model.
Can the common cold protect you from influenza? A murine model of respiratory viral co-infection.
Authors: Gonzalez Andres , Clary John , Rodgers Jade , Rodriguez Anna , Miura Tanya 
Published: January 2023

Abstract
Clinical cases demonstrate that up to 30% of respiratory infections involve more than one virus with rhinovirus being one of the most common in co-infections. Interestingly, rhinovirus has been found to exacerbate disease when co-infecting with adenovirus, whereas it can modulate disease when co-infecting with influenza A virus. However, there has yet to be a clear connection for how these viruses interact in the host to alter immune responses and disease severity. We are using a murine model of respiratory viral co-infection to determine how minor group rhinovirus (RV1B) affects pathogenesis during influenza A virus (PR8) infection. BALB/c mice were infected with RV1B or mock-infected two days before infecting with a high (100% lethal) or low (40% lethal) dose of PR8. Mice co-infected with RV1B and low dose PR8 had reduced clinical signs, weight loss and mortality compared to mice infected with PR8 alone. However, RV1B did not reduce disease severity upon infection with a high dose of PR8. Groups of mice were infected with RV1B or mock-infected two days prior to infection with low dose PR8 and were sacrificed 4 and 7 days later to quantify PR8 in the lungs. Single- and co-infected mice had similar PR8 titers on day 4; however, the co-infected group completely cleared PR8 on day 7 while the single infected group still had detectable titers. These results suggest that rhinovirus stimulates an immune response that promotes viral clearance and recovery upon infection with influenza A virus. Ongoing work will characterize this response to explain how respiratory viral co-infections can manipulate host immune responses to change disease outcome.
Memory CD8 T cells in RSV infection: friend or foe?
Authors: Stoley Megan E , Knudson Cory J , Pewe Lecia L , Meyerholz David K , Langlois Ryan A , tenOever Benjamin R , Harty John T , Varga Steven M 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is the leading cause of severe respiratory tract infection in infants and young children. CD8 T cells play a critical role in mediating viral clearance following an acute RSV infection. However the role of memory CD8 T cells in providing protection against RSV remains understudied. To generate high magnitude CD8 T cell memory in the absence of CD4 T cell memory and antibodies, we immunized naïve mice with dendritic cells pulsed with an RSV-derived peptide followed by a boost with a recombinant Listeria monocytogenes expressing the same RSV-derived epitope. Memory CD8 T cells significantly reduced viral titers following RSV challenge, but did so at the expense of increased airway dysfunction, weight loss, and mortality compared to controls. Importantly, the severe immunopathology and mortality observed was specific to the context of an RSV infection, as prime-boosted mice challenged with a recombinant influenza virus expressing the same RSV-derived epitope did not exhibit enhanced disease. The induction of a pro-inflammatory cytokine storm mediated by TNF-α and IFN-γ was observed in the serum of prime-boosted mice following RSV challenge. Additionally, RSV-specific memory CD8 T cells produced large amounts of IFN-γ locally within the lung, and adoptive transfer of wild-type but not IFN-γ-deficient memory CD8 T cells resulted in enhanced airway dysfunction and weight loss. Our results indicate that memory CD8 T cells are able to mediate protection against RSV infection. However, memory CD8 T cells acting alone in the absence of antibodies and memory CD4 T cells induce significant immunopathology and mortality through the induction of a systemic pro-inflammatory cytokine storm and local IFN-γ production.
Differential antigen presentation and transport by neonatal and adult dendritic cell subsets during Respiratory Syncytial Virus (RSV) infection
Authors: Lau-Kilby Annie W , Kehl Margaret , Malloy Allison MW 
Published: January 2023

Abstract
Dendritic cells (DCs) are critical for the induction of anti-viral CD8+ T cells. Young infants mount poor antiviral responses contributing to increased disease susceptibility and severity compared with acquisition later in life. Using a mouse model of respiratory syncytial virus (RSV), we have previously shown differences in the number of DCs that can carry antigen from the lung to the lung-draining lymph node (dLN) during infection between adults and neonates, and this is associated with age-dependent differences in RSV-specific CD8+ T cell responses. However whether adult or neonatal DCs more readily present antigen for T cell engagement is unknown.
In this study, we used an ovalbumin-expressing RSV (RSV-OVA) to track antigen presentation through early infection. In the lung, expression of MHCI-bound OVA (MHCI-OVA) was highest on adult CD103+ DCs; while in the dLN a greater number of neonatal CD103+ DCs expressed MHCI-OVA. Quantification of the migration-associated chemokine receptor 7 identified increased expression on neonatal MHCI-OVA-expressing CD103+ DCs compared to adults, suggesting adults may more readily present antigen but neonatal CD103+ DCs were more apt to migrate upon antigen acquisition. However, neonates failed to induce an OVA-specific T cell response in the lung in contrast to adults.
Together our data indicate that in neonates MHCI-OVA+ CD103+ DCs were more proficient at migration resulting in a greater number of MHCI-OVA-presenting CD103+ DCs in the dLN;- however, they failed to induce an OVA-specific T cell response. Thus antigen abundance was not the primary determinant of CD8+ T cell induction during RSV infection in neonates, suggesting that alternate mechanism(s) limit their DC-mediated T cell responses.
Tetherin controls T cell proliferation and exhaustion by shaping the early distribution of a persistent viral infection
Authors: Kenyon Elizabeth A , Nayak Debasis , McGavern Dorian B 
Published: January 2023

Abstract
Type I interferons participate in the host defense against most viral infections through induction of innate antiviral proteins like tetherin (BST-2). In vitro studies have shown that tetherin can physically sequester viruses by linking them to infected cells, which in turn reduces virion release. It is not currently understood how this antiviral protein functions in vivo to control an immune response mounted against a persistent viral infection. We sought novel mechanistic insights into the function of tetherin by studying its role in the establishment of a persistent lymphocytic choriomeningitis virus (LCMV) infection. Following viral infection, tetherin was significantly upregulated in B cells, plasmacytoid dendritic cells (DCs), conventional DCs, and brain-resident microglia. Genetic deletion of tetherin resulted in an early failure to contain a persistence-prone strain of LCMV in the splenic marginal zone. This resulted in a rapid re-distribution of LCMV into the red and white pulp at early time points post-infection. This also interfered with the early priming of antiviral T cells by causing them to localize preferentially to the splenic red pulp where they proliferated less and acquired an impaired phenotype characterized by upregulation of exhaustion markers (PD-1, LAG-3, TIGIT), loss of cytokine production, and decreased motility. Importantly, these early alterations impeded the ability of the host to control persistence-prone LCMV. Viral control was delayed in peripheral tissues, and lifelong persistence was established in the brain. These data demonstrate that tetherin plays a critical in vivo role in shaping the anatomical distribution and adaptive immune response against a persistent viral infection.
Transposon mutagenesis screen identifies host-encoded anti-viral factors
Authors: Bruchez Anna M , Sha Ky , Chen Li , Lacy-Hulbert Adam , Stuart Lynda 
Published: January 2023

Abstract
Ebola virus causes sporadic outbreaks of severe hemorrhagic fever. Owing to the high mortality rate, it poses a significant health risk and is a potential bioterrorism agent. However, our knowledge of how it usurps host cells to complete its life cycle, and more importantly, how this can be combated, is limited.
Here we describe a genome-wide forward genetic approach to identify host genes, known as restriction factors, which confer resistance to viral infection. In this system cells are mutagenized with a transposon that activates genes close to its insertion site. Libraries of mutagenized cells are screened with a cytotoxic virus and resistant cell populations are selected. Illumina sequencing is used to characterize transposon insertion sites and identify candidate resistance genes.
We have performed this screen in five human cell lines to identify genes conferring resistance to either vesicular stomatitis virus (VSV) or recombinant VSV bearing the Ebola virus surface glycoprotein. We have identified genes associated with resistance to only Ebola-VSV, or to both viruses, demonstrating that resistance occurs by targeting either viral entry or viral replication. One of the genes our screen identified is NPC1, the Ebola virus entry receptor, thus confirming that this is a valid method of identifying virus-host interactions. We have also identified a number of novel candidate Ebola restriction factors, which we are characterizing further, and are extending the screen to additional cell lines and viruses. This work provides new insights into host anti-viral defense mechanisms and may identify novel therapeutic targets that can be utilized to promote host-encoded anti-viral immunity.
Mutations in the 5′ NTR and the non-structural protein 3A of the coxsackievirus B3 selectively attenuate myocarditogenicity
Authors: Massilamany Chandirasegaran , Gangaplara Arunakumar , Basavalingappa Rakesh Halekote , Rajasekaran Rajkumar A , Steffen David , Pattnaik Asit K , Reddy Jay 
Published: January 2023

Abstract
The 5′ non-translated region (NTR) is an important molecular determinant that controls replication and virulence of coxsackievirus B (CVB)3. In our studies of CVB3-induced myocarditis, we sought to generate an infectious clone of the virus for routine in vivo experimentation. By determining the viral nucleotide sequence, we identified three new nucleotide substitutions in the clone that differed from the parental virus strain: C97U in the 5′ NTR; a silent mutation, A4327G, in non-structural protein 2C; and C5088U (resulting in P1449L amino acid change) in non-structural protein 3A of the virus leading us to evaluate the role of these changes in the virulence properties of the virus. We noted that the disease-inducing ability of the infectious clone-derived virus in three mouse strains was restricted to pancreatitis alone, and the incidence and severity of myocarditis were significantly reduced. We then reversed the mutations by creating three new clones, representing 1) U97C; 2) G4327A and U5088C; and 3) their combination together in the third clone. The viral titers obtained from all the clones were comparable, but the virions derived from the third clone induced myocarditis comparable to that induced by wild type virus; however, the pancreatitis-inducing ability remained unaltered, suggesting that the mutations described above selectively influence myocarditogenicity. Our data point to a possibility that, CVB3 viruses containing such altered nucleotides may evolve naturally leading to their survival.