bluuebunny/crossref_metadata_2025_split huggingface dataset:
TLR4-Deficient C3H/HeJ Mice Exhibit Enhanced Disease and Mortality Following Intranasal MHV-1 Infection (44.23)
Authors: Haag Brayton A , Hartwig Stacey M , Khanolkar Aaruni , Harty John T , Varga Steven M 
Published: January 2023

Abstract
Severe acute respiratory syndrome (SARS) is characterized by substantial acute pulmonary inflammation and a high mortality rate. A novel human coronavirus, SARS-associated coronavirus (SARS-CoV), was identified as the etiologic agent of SARS. With the disappearance of SARS in the human population following the initial outbreak in 2003, limited information is available regarding the immunopathogenesis of SARS. Thus, an animal model that recapitulates the human disease would prove to be of great benefit, providing valuable insight into the pathology of the infection. Recent studies have demonstrated that intranasal infection of susceptible mouse strains with a related mouse coronavirus, mouse hepatitis strain 1 (MHV-1), produces acute respiratory disease and mortality that resembles human SARS cases. We find that intranasal infection with MHV-1 results in morbidity and some mortality in susceptible C3H/HeN (H-2k) mice. Interestingly, MHV-1 infection of C3H/HeJ mice, that harbor a natural mutation in their toll-like receptor 4 (TLR4) gene which renders TLR4 unable to properly signal, results in enhanced disease and increased mortality. In the absence of a functional TLR4 signal, we observe enhanced CD4 T cell responses, where as CD8 T cell responses remain unaltered. Our results indicate that TLR4 plays an important role in respiratory coronavirus pathogenesis.
Mast cells induce alternative activation and inhibit caspase-3 mediated apoptosis of Francisella tularensis infected macrophages (44.24)
Authors: Rodriguez Annette Rose , Yu JiehJuen , Murthy Ashlesh , Guentzel M. Neal , Klose Karl E. , Chambers James P. , Forsthurber Thomas G. , Berton Michael T. , Arulanandam Bernard P. 
Published: January 2023

Abstract
Francisella tularensis is a highly infectious Gram-negative bacterial pathogen and the causative agent of pneumonic tularemia. We recently demonstrated inhibition of F. tularensis LVS replication in macrophages via mast cell production of interleukin-4 (IL-4). We have further analyzed the interactions of mast cells and macrophages using a primary bone marrow derived mast cell-macrophage co-culture system and determined that mast cells inhibit F. tularensis-induced macrophage apoptosis. Specifically, mast cells co-cultured with macrophages significantly reduced intramacrophage LVS growth and expression of apoptotic proteins caspase-3 (19.9%) and PARP (Poly-ADP ribose polymerase, 16.0%) compared to macrophages cultured alone (48.2% and 56.4%, respectively). The inhibition of LVS-induced apoptosis within infected macrophages also was seen by the direct addition of recombinant IL-4 and corresponded with up-regulation of mannose receptor expression and arginase activity which are associated with the alternative activation of macrophages. Moreover, IL4R-/- mice were more susceptible to intranasal challenge and exhibited greater caspase-3 activity in lung macrophages (12.9%) compared to similarly challenged wild-type (4.7%) animals. These results support a role for mast cells and IL-4 in control of bacterial replication and apoptosis following pulmonary LVS infection.
National Institutes of Health grant PO1 AI057986; NIH/NIGMS MBRS-RISE GM60655
Protection of cynomolgus macaques from Ebola Sudan infection (45.10)
Authors: Dye John M , Muhammad Majidat , Bakken Russell , Barth James , Kuehne Ana , Olinger Gene G 
Published: January 2023

Abstract
Ebola virus, a filovirus, is a highly pathogenic human pathogen for which there is no vaccine or treatments licensed for human use. The Sudan subtype of the Ebola virus (SEBOV) was first identified in 1976 after a hemorrhagic fever outbreak in that region of Africa. There were reemergences of the SEBOV in sporadic outbreaks from 2000 to 2004 with a 40-65% case fatality rate in humans. In this study, we evaluated the use of the Venezuelan equine encephalitis replicon (VRP) vaccine vector expressing the glycoprotein (GP) of Ebola Sudan to protect cynomolgus macaque nonhuman primates (NHP) against a uniformly lethal challenge with SEBOV. We found that six of six NHP vaccinated with VRP expressing GP of Sudan were protected against infection with 1000 PFU of SEBOV. Furthermore, when we back challenged these Sudan survivors with another strain of Ebola, Zaire, there was only a partial protection (two of six survived). When we vaccinated NHP with VRP expressing GP of Sudan and GP of Zaire, we found that all animals were protected from either infection with Sudan or Zaire (three of three survived both infections). Taken in total, these data indicate that vaccination with only one subtype of Ebola virus will not protect against other subtypes. However, there is no interference when using multiple VRP vaccinations against different subtypes of Ebola viruses. These results are encouraging for the development of a multivalent vaccine platform against multiple strains of filoviruses.
The antigen-specific CD4 host response to Plasmodium yoelii 17XL infection after protective MSP-8 immunization alters multifunctional cytokine profiles (45.13)
Authors: Smigiel Kate , Darrah Patricia , Fogg Chris , Harvey Reid , Tullo Greg , Seder Robert , Burns James , Long Carole 
Published: January 2023

Abstract
Multifunctional CD4 T cells, which produce combinations of interferon gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), and IL-2, have been shown to be immune correlates of protection after vaccination in a mouse model of L. major and for some models of M. tuberculosis. In this study, we have used multi-parameter flow cytometry to study a merozoite-surface protein-based (MSP-8) vaccine model against lethal murine Plasmodium yoelii (Py) infection, which we have shown to be dependent on IFN-γ and which also requires an active immune response during challenge infection for protection and survival. Our data supports previous studies, which have found that the patterns of multifunctional responses are established after the first immunization but the frequencies of CD4 responders and the amount of IFN-γ produced per multifunctional cell increase with subsequent immunizations. We further show that antigen-specific memory CD4 cells in the lymph nodes are functionally distinct from those in the spleen and liver before and after Py challenge. Also, the nonlymphoid-tissue (liver) responses occur at the greatest frequency and show the greatest plasticity in cytokine profile during infection. We conclude that this protective immunization induces multifunctional 3+ antigen-specific CD4 T cells (10-25% of MSP-8 specific CD4s) but that the host response to lethal malaria infection triggers a shift in multifunctionality towards more IFN-γ production, especially in non-lymphoid tissue.
Funding Sources: J. Burns (NIH #AI35661), C. Long (Intramural Program of NIH-NIAID)
Cytolytic Potentials of CD8+ T Cells Persisting in Genital Skin and Mucosa after HSV-2 Reactivation (45.16)
Authors: Zhu Jia , Phasouk Khamsone , Peng Tao , Woodward Amanda , Klock Alexis , Corey Lawrence 
Published: January 2023

Abstract
We have previously demonstrated that CD8+ T cells, including HSV-2 specific CD8+ T cells, persisted in genital skin for months after the herpes lesion completely healed, and preferentially accumulated at the dermal-epidermal junction where sensory nerve endings were enriched. These persisting CD8+ T cells have been postulated in the containment of the reactivating virus before clinical symptomatic lesion occurs. To understand functions of persisting CD8+ T cells in human, we have combined immunofluorescent (IF) staining, laser capture microdissection (LCM) and RT-PCR to analyze the gene expression of CD8+ T cells at the dermal-epidermal junction in post-healed lesion biopsy. CD8+ T cells were captured at a single cell level. The purity was validated by the lack of CD4 and a 100-fold more enriched CD8 expression in LCM captured cells than in whole tissue sections. In captured CD8+ T cells, we detected up-regulation of genes encoding cytolytic granules, such as perforin and granzyme B, as compared to keratinocytes in the epidermis. The expression of cytolytic granules was further confirmed by IF staining of persistent CD8+ T cells, even in post-healed biopsy of acyclovir treated individual. Our data strongly indicated the cytolytic potentials of CD8+ T cells persisting at the dermal-epidermal junction and implicated their role in immune surveillance against HSV-2 reactivation in skin and mucosa.
This work was supported by NIH grants R37 AI042528 and PO1 AI030731.
IL-7 and IL-15 play a synergistic role in the development of CD8+ T cell response against an obligate intracellular parasite (45.17)
Authors: Bhadra Rajarshi , Guan Hongbing , Khan Imtiaz A. 
Published: January 2023

Abstract
IL-7 and IL-15 are believed to be important for homeostatic proliferation of CD8+ memory T cells. However, the simultaneous role played by these two cytokines in the generation of this response during acute phase of infection has not been described. Our results demonstrate that depletion of IL-7 in mice lacking IL-15 gene leads to poor development of both short-lived effectors as well as memory phenotype CD8+ T cells during Toxoplasma gondii infection. CD8+ T cells from these animals lack the ability to produce optimal levels of IFNγ upon antigenic-restimulation and their ability to exhibit cytotoxic response is severely hampered. Moreover, Bcl-2 and KI-67 expression on these cells is down regulated. To the best of our knowledge this synergism between IL-7 and IL-15 in generating a potent CD8+ T cell immunity in an infectious disease model has not been previously reported.
Differential immunomodulatory activity of soluble beta glucans from barley and yeast in antigen specific humoral immune responses (45.21)
Authors: Bagriacik Emin Umit , Uslu Kadriye , Imir Turgut 
Published: January 2023

Abstract
The main difference in chemical structure of (1 →3)-β-D-glucans from yeast cell wall and barley is the branching side chains. Yeast cell wall contains (1→3),(1→6)-β-D-glucans whereas barley contains (1→3),(1→4)-β-D-glucans. The purpose of this study was to compare adjuvant effects of two soluble beta-glucans from barley and yeast (Saccharomyces cerevisiae) in antigen specific humoral immune responses. In a conalbumin-immunized mouse model, mice were immunized by conalbumin at a relatively lower concentration (10 μg/mouse) in the presence of beta glucans. Anti-conalbumin antibodies in the serum specimens of immunized and control mice were quantified by ELISA.
At high doses, both glucans were effective in increasing circulating conalbumin-specific antibodies of IgM, IgG, and IgA types. However, 1 μg of â-glucan from barley resulted in lower yield in IgG1, IgG2a, IgG2b, and IgGA levels in comparison to the yeast-derived glucan at the same dose. We also found that antigen specific antibody levels enhanced by the β-glucan from yeast were always higher than those of the glucan from barley.
In conclusion, yeast-derived glucans might have better immunomodulatory or adjuvat activity in induction of antigen specific humoral immune responses over barley-derived glucans.β
The role of IL-12p40 during host cell infection with Yersinia pestis KIM D27 (45.22)
Authors: Parent Michelle A. , Joerger Torsten A. , Golwala Sohil N. , Smiley Stephen T. , Blumerman Seth L. 
Published: January 2023

Abstract
Yersinia pestis, a facultative intracellular pathogen, is the causative agent of plague. A previously developed vaccine model using Yersinia pestis KIM D27, B cell-deficient mice, and convalescent serum resulted in the generation of T cell-meditated protection against lethal pneumonic infection. In order to understand how this serum results in the development of protective T cells we investigated the interaction between the host immune response, convalescent serum and Y. pestis. First, we determined that IL-12p40 but not IL-12p35 was required for mice to survive this vaccination regimen. We then began to examine the early host response to infection by studying dendritic cells (DC) infected with Y. pestis in the presence of convalescent serum, since DC are cable of presenting antigen to naïve T cells. We infected cultures of bone marrow-derived DC and established that Y. pestis is able to infect DC. Interestingly, convalescent serum does not result in clearance of organisms but sustains infection. Currently, we are investigating cytokine production and cell surface maker expression profiles in Y. pestis infected DC. Preliminary data of infected DC in the presence of convalescent serum results in an increase in IL-12p40 gene expression. Our goal is to ascertain those events occurring early during the course of host infection that lead to generation of this cell-mediated protection. This research is supported by a grant from the University of Delaware Research Foundation.
Identification of novel HLA-A*0201-restricted CD8+ T-cell epitopes on Ebola Zaire glycoprotein (45.26)
Authors: Prugar Laura Irene , Rebekah James M , Gene Olinger G 
Published: January 2023

Abstract
Due to high morbidity and mortality rates, filoviruses are considered among the deadliest of human pathogens with case fatality rates that can reach 90% in some human outbreaks. Ebolavirus and Marburgvirus, the two genera of the family Filoviridae, pose a significant threat to military personnel, global security, and public health, and are classified as Category A bioterrorism agents by the CDC. While there is no licensed vaccine available, there are several promising vaccine candidates that have demonstrated immunogenicity and efficacy in animal models of disease. The vaccine candidates to date, have identified immunogens, usually the glycoprotein (GP), and most importantly,have demonstrated efficacy in the highly sensitive macaque models of filovirus disease. Human vaccine trials will need immunoassays that correlate with protection. Our lab has demonstrated the role of CTL in rodent models of Ebola Zaire. Here we report the first identified CTL epitopes of Ebola Zaire virus (EBOVZ) GP on HLA A*201. The binding of HLA-A*0201-restricted peptide in EBOVZ GP was determined by peptide binding alogorithiums, pentamer binding assays, and binding of overlapping or nonamers using T2 cells expressing HLA-A*0201. Potential HLA-A*0201-restricted peptides were selected from the SYFPEITHI database and screened by T2 cell-stabilization assay. In addition, overlapping peptides were used to identify potential epitopes that could be missed from the predicted algorithms. The various putative epitopes were winnowed and validated with T2-cell stabilization assays. Our results will provide a new insight into the development of therapeutic vaccines and the development of assays for future human vaccine trials for EBOVZ.
Dengue viral component triggering reactive nitrogen and oxygen species production leads to endothelial cell apoptosis (45.33)
Authors: Yen Yu-Ting , Wu-Hsieh Betty 
Published: January 2023

Abstract
Vascular leakage is one of the life-threatening complications occurring in dengue patients, however, the pathogenic mechanism is not well understood. In dissecting the complex interplay between the virus and the host, we found that dengue virus productively infects the endothelial cells and causes apoptotic cell death through triggering the production of reactive nitrogen (RNS) and oxygen radicals (ROS).
To determine the dengue virus component(s) that is responsible for the cause of cell death, we transfected human microvascular endothelial cell HMEC with pCR3.1 plasmids encoding Flag-tagged viral components Core, PrM, NS1, NS2A, NS2B, NS3, NS2B-NS3, NS4A, NS4B, and NS5, separately. Apoptotsis in cells transfected with NS2B-NS3 was 10-fold higher and that in cells transfected with NS1, NS3 and NS5 or core was only between 4- to 6-fold higher than cells transfected with vector alone. Confocal microscopy revealed that while NS2B expression remained in the cytosol, NS3 and NS2B-NS3 in both the cytosol and the nucleus at 24 h after transfection. Interestingly, the effect of NS2B-NS3 on endothelial cells was reversed in the presence of NO and ROS inhibitors. These results demonstrate that NS2B-NS3, being the major component that induces endothelial cell apoptosis, interacts with subcellular components in both the cytosol and the nucleus, and induces the production of RNS and ROS to cause apoptotic outcome.
CD4 T cell derived IFN-gamma protects from lethal encephalomyelitis by inhibiting neutrophil infiltration independent of IL-17 (45.6)
Authors: Savarin Carine , Bergmann Cornelia C , Stohlman Stephen A 
Published: January 2023

Abstract
Immunodeficient SCID recipients of virus-primed memory CD4 T cells survive CNS infection by neurotropic coronavirus (JHMV), whereas IFN-γ deficient (GKO) donor CD4 T cells substantially increase morbidity. Control of virus replication and demyelination are similar in both recipients compared to uncontrolled replication without demyelination in non-treated mice. However, distinct from wt CD4 T cell recipients, GKO recipients exhibit massive CNS neutrophil infiltration, prominent CD4 T cell localization to grey matter, and detection of IL-17 mRNA. To determine the relative role of IFN-γ, IL-17 and neutrophils in disease severity, equal populations of wt and GKO CD4 T cells were co-transferred to SCID mice. Despite the vast abundance of GKO versus wt CD4 T cells accumulating in the CNS at 8 days p.i. (80 vs 20 %), disease severity and survival was similar to protected recipients of wt CD4 T cells. Furthermore, the minor population of wt CD4 T cells in mixed recipients resulted in decreased CNS neutrophil infiltration, consistent with modestly elevated IFN-γ and reduced neutrophil-attracting chemokines. However, wt CD4 T cells did not reduce CNS IL-17 mRNA levels in recipients of mixed relative to GKO donor cells. The data suggest that IFN-γ dominates over IL-17 in controlling excessive neutrophil recruitment and thus protecting JHMV infected SCID recipients from premature death.
Determining the role of Ikaros in Th17 differentiation (46.18)
Authors: Wong Larry , Brown Melissa 
Published: January 2023

Abstract
CD4+ Th17 T cells contribute to the pathogenesis of a number of autoimmune/inflammatory diseases. Yet the molecular events that govern their development are still poorly understood. A reciprocal relationship between Th17 and T regulatory (Tregs) cells exists such that the relative expression of cell-specific transcription factors, RORγt and Foxp3, dictates whether CD4+ T cells differentiate into Th17 or Tregs cells, respectively. We have defined a role for Ikaros in these cell fate decisions. Ikaros is a site-specific DNA-binding transcription factor involved in the epigenetic regulation of several T cell cytokine genes. We demonstrate that Ikarosnull CD4+ T cells are developmentally impaired in vitro under Th17- skewing conditions. In addition to producing less IL-17, these cells exhibit reduced RORγt and higher Foxp3 expression than their wild type counterparts. Furthermore, all Foxp3+ T cells are also CD25+, suggesting that the lack of Ikaros predisposes Th17 T cells to a regulatory T cell phenotype. The effect of Ikaros on IL-17 production is not limited to CD4+ T cells. Ikarosnull CD8+ T cells also have dramatically lower IL-17 production. However, unlike Ikaros-deficient CD4 T cells, Ikarosnull CD8+ T cells do not express markers of Treg cells but show increased IFNγ production. Thus, our data suggest Ikaros is intimately involved in the differentiation of IL-17-producing T cells.
This research is supported by NIH RO1 NS047578
The mechanism of protection against influenza by transfer of Tc17, an IL-17 secreting CD8 T cell subset. (46.5)
Authors: Hamada Hiromasa , Garcia-Hernandez Maria de la Luz , Reome Joyce B. , Misra Sara K. , Dutton Richard W. 
Published: January 2023

Abstract
Interleukin (IL) -17 secreting CD4 T (Th)17 and CD8 T (Tc)17 effector cells are found in the lung following primary challenge with influenza A and blocking antibody to IL-17 increases weight loss and reduces survival. Tc17 effectors can be generated in vitro using naïve CD8 T cells from OT-1 T cell receptor (TcR) transgenic mice. T cell numbers expand 20 fold and a majority secretes IL-17, but little IFN-γ. Tc17 effectors are negative for granzyme B, for perforin message and lack cytolytic activity in vitro, in contrast to Tc1 effectors. Many of the Tc17 effectors become IFN-γ/IL-17 double producers after adoptive transfer and kill targets by a Fas dependent, perforin independent, pathway in an in vivo CTL assay. Adoptively transferred Tc17 cells expand greatly after transfer to naïve recipients following challenge and can protect them against lethal influenza infection. Transfer of Tc17 effector cell enhances the infiltration of neutrophils and monocytes to lung after influenza infection and protects against the development of respiratory distress much more effectively than transfer of equivalent numbers of Tc1 effectors. Additional mechansims of protection are being explored.
This work was supported by NIH AI046530 and the Trudeau Institute.
The effects of age on CD4 upregulation and Treg distribution in a TAChR peptide-specific immune response (47.1)
Authors: Stacy Sue , Standifer N. E. , Pasquali Amanda , Krolick K. A. , Infante A. J. , Kraig Ellen 
Published: January 2023

Abstract
Studies of age-related defects in T cell function have been complicated by the difficulty of identifying antigen-specific T cells early in the activation process. Thus, peptide-responsive T cells were identified and characterized by the upregulation of CD4, an early event following T cell receptor ligation. We focused on the Torpedo californica acetylcholine receptor (TAChR) peptide, p146-162, since responses to this peptide have been well defined in young mice. In old mice, the fine specificity of the TAChR response was preserved with age; p146-162 remained immunodominant. Yet, in vitro proliferative responses to p146-162 were diminished in a dose-dependent manner in lymph node cells (LNC) derived from old peptide-primed mice. When p146-162-specific Vβ6+CD4high T cells derived from young (2-5 months) and old (20-27 months) mice were examined, the upregulation of CD4 was maintained in old age but the frequency of Vβ6+CD4high T cells was often lower; reflecting fewer CD4+ LNC, changes in TCR repertoire selection and/or altered T cell regulation. Interestingly, Foxp3+CD25+CD4+ T regulatory cell (Treg) frequencies were elevated in the old peptide-responding populations. The Vβ6-CD4normal Treg phenotype predominated in both age groups, yet a unique "activated" CD4high Treg subpopulation was also identified. Future studies may help elucidate the potential role of these CD4high Tregs in promoting age-related immune dysfunction. Support: Myasthenia Gravis Foundation, Nathan Shock Aging Center, NIH R03 AG 14557 and San Antonio Area Foundation.