bluuebunny/crossref_metadata_2025_split huggingface dataset:
FAK regulates progenitor B cell growth and retention in their bone marrow microenvironments (111.39)
Authors: Park Shin-Young , Wolfram Peter , Manis John , Beggs Hilary , Nombela-Arrieta Cesar , Pivarnik Gregory , Silberstein Leslie 
Published: January 2023

Abstract
Progenitor B cells reside in complex bone marrow microenvironments where they receive signals for growth and maturation. Here we have conditionally targeted in B cells Focal Adhesion Kinase (FAK), an integrator of cell extrinsic signals including CXCL12 and VCAM-1. The numbers of progenitor (pro-B and pre-B) and immature B cells are reduced by 30-40% in CD19-Cre Fakfl/fl mice. When cultured in methylcellulose with IL-7 ± CXCL12, Fak deleted pro-B cells undergo increased cell division and cell cycle progression, which correlates with down-regulation of four cell cycle regulatory genes, i.e. Cdk5rap1, Cdkn2a, E2f2, and Sfn. However, the increased cell division is accompanied by increased apoptosis, ultimately yielding fewer B cells and colonies. Utilizing quantitative laser scanning cytometry of longitudinal sections of femoral bone marrow, we show that Fak deletion disrupts the non-random gradient distribution of pro-B cells, which are preferentially localized in endosteal regions of the metaphyses and diaphysis. Increased numbers of pro-B cells are detected in the periphery of Fak deleted mice, and mobilization of pro-B cells induced by the inflammatory antigen NP-CGG-alum is increased 3-fold in Fak-deleted versus wild type mice. Collectively, these studies illustrate the importance of FAK in regulating progenitor B cell homeostasis and maintenance of their spatial distribution in bone marrow niches.
The AP-1 transcription factor Fra-1 is a negative regulator of B2B cell proliferation and of immunoglobulin class switch (111.49)
Authors: Groetsch Bettina , Lang Christiane , Neubert Kirsten , Brachs Sebastian , Zech Christine , Derer Anja , Luther Julia , Voehringer David , Wagner Erwin , Schett Georg , Mielenz Dirk , David Jean-Pierre 
Published: January 2023

Abstract
AP-1 transcription factors are immediate early genes that sense changes in the cellular environment to control differentiation, proliferation and apoptosis. Their functions in B cells are poorly described and were only suggested by the pro-apoptotic role of JunB. Fra-1(FosL1), an interaction partner of JunB, has been firstly described in activated B cells. Here, we confirm that Fra-1 becomes up-regulated in activated murine primary B cells that prompted us to analyze B cell development and humoral immune responses following Fra-1 gain and loss of function in vivo. Mice over-expressing Fra-1 that displayed a reduced B2 B cell compartment failed to mount any IgG response and did not form germinal centers. Bone marrow transfer in mice lacking B cells demonstrated that these phenotypes were B cell autonomous. B cell-specific Fra-1 deficient mice developed enhanced secondary responses, further reinforcing a cell autonomous key role for Fra-1 in B cell function. In vitro experiments showed that sorted fra-1tg B2 cells that express all relevant genes required for plasma cell differentiation and class switching were unable to class-switch, do not proliferate and die after activation. Thus, we identified Fra-1 as negative regulator of IgG production and class switch that functions independently of chromatin remodeling and class switch gene expression. Our work therefore suggests that Fra-1 acts as a negative regulator of B2 B cell proliferation, which is required for Ig class switch.
All-trans retinoic acid-induced antimicrobial activity against M. tuberculosis is dependent on NPC2-mediated regulation of cellular cholesterol (112.1)
Authors: Liu Philip , Kim Elliot , Wheelwright Matthew , Krutzik Stephan 
Published: January 2023

Abstract
One key function of the innate immune response is the rapid detection and elimination of microbial invaders, which can be influenced by micronutrients. A role for vitamin A in the host defense against Mycobacterium tuberculosis has been suggested through epidemiological and in vitro studies; however, the antimicrobial mechanism remained unclear. Here, we demonstrate that treatment of M. tuberculosis-infected human monocytes with either all-trans retinoic acid (ATRA) or 1,25-dihydroxyvitamin D, the biologically active forms of vitamin A and vitamin D respectively, resulted in antimicrobial activity against the pathogen. Comparison of the ATRA and 1,25D3 elaborated gene expression profiles revealed that ATRA distinctly triggered a lipid metabolism and efflux pathway in human monocytes. Accordingly, stimulation with ATRA but not 1,25D3 led to decreased cellular cholesterol content and altered intracellular lipid distribution, which were both dependent upon induction of NPC2. Knockdown of NPC2 expression resulted in the loss of ATRA- but not 1,25D3-induced antimicrobial activity against M. tuberculosis. These results demonstrate that ATRA-triggered host defense is mediated through NPC2-dependent regulation of cellular cholesterol content.
Lipocalin 2 mediated hypoferremia protects against endotoxin- induced sepsis in mice (112.11)
Authors: Vijay-Kumar Matam , Aitken Jesse , Srinivasan Gayathri , Sashidharamurthy Rangaiah , Jones Dean , Gewirtz Andrew 
Published: January 2023

Abstract
Sepsis is characterized by hypoferremia, a primitive protective mechanism to limit iron-induced oxidative stress. Lipocalin-2 (Lcn2) which is highly upregulated during inflammation and has been shown to transport iron under physiologic conditions. Accordingly, we hypothesized that such upregulation of Lcn2 serves to protect against sepsis associated oxidative stress. Lcn2 deficient mice (Lcn2KO) and their WT littermates were given E. coli LPS (20mg/kg BW) i.p. and monitored for mortality. Serum TNFα, IL-18, LDH, aminotransferases, iron, glutathione and cysteine were analyzed. Splenocyte apoptosis was assayed by TUNEL, caspase 3 cleavage and flow cytometry. In WT mice, LPS-induced systemic Lcn2 peaked at 24h and returned to basal levels by 48h. All Lcn2KO died by day 8 while only 20% mortality was observed in WT mice. In addition, Lcn2KO exhibited substantial elevation of every parameter tested including both pro- and anti-inflammatory cytokines. Further,Lcn2KO splenocytes exhibited extensive apoptosis as measured by caspase 3 cleavage, TUNEL staining and flow cytometry. More importantly, when compared to WT mice, Lcn2KO mice failed to exhibit LPS-induced hypoferremia at early time points and exhibited severe hepatic oxidative stress. Our data demonstrate that Lcn2 facilitates LPS-induced hypoferremia and serves as a host protective factor by reducing oxidative stress. Given Lcn2 multifunctionality, it may be developed as a potential therapeutic agent to treat sepsis.
Epithelial SPLUNC1 mediated antimicrobial activity against Klebsiella pneumoniae (112.14)
Authors: Di Y.Peter , Birru Rahel 
Published: January 2023

Abstract
Short PLUNC protein 1 (SPLUNC1) is an epithelial antimicrobial protein that is highly expressed in the nasal, pharyngeal, and lung epithelia. We have recently shown that in a transgenic mouse model (Scgb1a1-hSPLUNC1) over-expressing human SPLUNC1 is critical in pulmonary innate immune response against P. aeruginosa. The objective of this study was to identify differentially expressed transcripts using microarray analysis to understand the genetic basis of how SPLUNC1 regulates innate immune response in host against Klebsiella pneumoniae, an insidious organism that causes invasive respiratory infections. Scgb1a1-hSPLUNC1 transgenic (TG) mice were challenged with K. pneumoniae and displayed significant resistance to bacterial infection than wild-type (WT) mice. Categorization of the differentially expressed genes showed that those preferentially involved in host immune responses were activated extensively and TG mice exhibited more robust response at early time points after infection than WT mice. Ingenuity pathway analysis results suggested several key inflammatory regulatory genes are regulated by the over-expression of hSPLUNC1. TG mice displayed enhanced bacterial clearance that was accompanied by a decrease in neutrophil infiltration and cytokine levels that resulted in enhanced mouse survival against K. pneumoniae-induced respiratory infection. Our data indicate that SPLUNC1 is a crucial component of mucosal immune defense against pulmonary infection with K. pneumoniae.
Pathogen array reveals innate immune defense against molecular mimicry (112.16)
Authors: Stowell Sean , Arthur Connie , McBride Ryan , Smith David , Paulson James , Cummings Richard 
Published: January 2023

Abstract
The microbiome regulates diverse biological processes, including general metabolism and immunity, within a given individual. However, the examination of immunological factors responsible for regulating this superorganism remains difficult. In order to overcome these current limitations, we developed a microbial glycan microarray from a diverse range of microbial species. Serological comparison between humans and mice demonstrated striking differences in reactivity toward the antigenic determinants of different species, strongly suggesting unique exposure and/or distinct immunological responses to microbial flora between organisms. More importantly, innate immune lectins, galectin-4 and galectin-8, displayed exquisite specificity for bacteria displaying self-like antigens, while failing to bind related strains with non-self-like antigens. Galectin binding to these bacteria caused death accompanied by alterations in membrane integrity, whereas binding to mammalian cells expressing similar structures had no effect on cell viability. These results demonstrate that microbial glycan microarrays provide a new platform to explore the distinct contributions of different immunological factors to a wide variety of organisms and strongly suggest that innate immune lectins exist that provide unique protection against microbial molecular mimicry.
Role of microRNAs in immune response to infection (112.18)
Authors: Mehta Manali , Liu Philip , Teles Rosane , Modlin Robert 
Published: January 2023

Abstract
Regulation of microRNAs has been shown to be critical in immune cell development as well as the progression of certain cancers. However, their role in immune response to microbial infection has remained elusive. We chose to use Mycobacterium leprae, the causative agent of leprosy, as a model to determine the potential roles of microRNA in pathogenesis. The immune response to M. leprae has been well-characterized as presenting as a spectrum, ranging from a more contained Th1 response in tuberculoid patients to a more disseminated Th2 response in lepromatous patients, providing a good system for examining the correlation of microRNAs and type of immune response. We performed mRNA and microRNA microarray analysis on 6 T-lep and 6 L-lep lesions to determine whether microRNA disregulation and subsequent downregulation of key immune genes contributes to the outcome and morbidity of infection. We discovered 7 microRNAs in T-lep and 28 microRNAs in L-lep lesions and cross-referenced them in MiRNA and Genes Integrated Analysis (MAGIA) web tool with mRNA gene transcripts that were significantly downregulated in the same lesions. Preliminary analysis of Th1 and Th2 gene targeting shows microRNAs more highly expressed in L-lep lesions may preferentially target proinflammatory genes, which could explain the bias in immune response to infection.
Small-molecule BET inhibitor JQ1 subdues inflammation in vitro and protects mice from LPS-induced shock (112.19)
Authors: Belkina Anna , Denis Gerald 
Published: January 2023

Abstract
New treatment strategies are greatly needed for both acute and chronic inflammatory diseases. A novel triazolediazepine-fused ring compound JQ1 was recently reported to disrupt interactions between acetylated chromatin and dual bromodomains of BET proteins Brd4 and Brd2. Brd2 knockdown mice demonstrate reduced inflammatory responses, and various Brd2 shRNA-expressing cells cannot fully respond to inflammatory stimuli. Our HYPOTHESIS was that JQ1 disrupts BRD interactions to ablate inflammatory responses in a similar manner. RESULTS. We treated RAW264.7 cells and BMDMs with JQ1, stimulated them with LPS, and assayed mRNA of cytokine genes and cytokine levels in culture medium (TNFa, IL-6, IL-10, MCP-1). We stimulated mouse B cells with E. coli LPS and Pam3CSK4 to see the effect of JQ1 on TLR-mediated response. Finally, we subjected C57BL/6 mice to a lethal dose of LPS and treated them with 50 mg/kg of JQ1. We recorded survival and measured cytokine response in the serum 20,40,60,90,120, and 240 min after LPS injection. We found that JQ1 treatment dramatically reduces inflammatory responses in vitro and in vivo, downregulating expression and production of key NFkB-dependent cytokines. In B cells, JQ1 reduced proliferative effects of TLR2 and TLR4 stimulation. 90% of JQ1-treated mice survived endotoxic shock while 0% survived in the non-treated group. JQ1 ablated serum inflammatory cytokines within 1 hr. Thus JQ1 is a potent, novel, epigenetically-acting anti-inflammatory.
SLAM is a microbial sensor which regulates bacterial phagosome functions in macrophages by recruiting a Vps34/beclin 1/UVRAG complex (112.8)
Authors: Ma Chunyan , Wang Ninghai , Detre Cynthia , Wang Guoxing , O’Keeffe Michael , Terhorst Cox 
Published: January 2023

Abstract
Phagocytosis is a pivotal process by which macrophages eliminate microorganisms upon recognition by pathogen sensors. Surprisingly, the self-ligand cell surface receptor Slamf1 functions not only as a co-stimulatory molecule but also as a microbial sensor controlling killing of Gram-negative bacteria by macrophages. Slamf1 regulates activity of the Nox2 enzyme and phagolysosomal maturation upon entering the phagosome, by recruiting the enzyme Vps34, which produces PI3P. Here we report that in mouse macrophages, Slamf1 interacted with Vps34, Beclin 1 and UVRAG in E. coli-containing phagosomes. However this interaction was also found without induction of phagocytosis in macrophages, transfectant HEK293 cells and human chronic lymphocytic leukemia cells. Removal of the second ITAM motif in the cytoplasmic tail of Slamf1 seemed to enhance the interaction of Slamf1 with Beclin 1, while Slamf1 tailless totally abolished the interaction. Both the BD and CCD domains of Beclin 1 were required for efficient binding to Slamf1. Slamf1 did not interact with Atg14L or Rubicon, which also complex with Vps34 and Beclin 1 in autophagy. We conclude that Slamf1 recruits a subset of Vps34 associated proteins, which is involved in membrane fusions.
Naturally processed HLA class I-restricted epitopes inform targets of protective CD8+ T cell-mediated immunity (113.11)
Authors: Gilchuk Pavlo , Spencer Charles , Conant Stephanie , Niu Xinnan , Erickson John , McAfee Kristi , Oseroff Carla , Hadrup Sine , Bennink Jack , Hildebrand William , Edwards Kathryn , Crowe, Jr James , Williams John , Buus Søren , Sette Alessandro , Schumacher Ton , Link Andrew , Joyce Sebastian 
Published: January 2023

Abstract
Rational design of CD8+ T cell (TCD8)-based vaccines requires knowledge of the immunogenic and protective epitopes presented during infection, information which is currently lacking. Using the clinically successful smallpox vaccine as a model, ~200 novel naturally processed vaccinia viral peptides presented by HLA-A*0201 and -B*0702 molecules were identified and characterized. Humans showed a variegated response to these determinants, reminiscent of the hierarchic response seen in HLA class I transgenic mice. Importantly, multiple TCD8 epitopes were commonly recognized by humans and mice and identified potential targets for protective TCD8-mediated immunity. After acute infection, both dominant and subdominant TCD8 specificities exhibited all of the immunologic features necessary for protection. However, early and efficient presentation of immune determinants during infection ensured protective responses, regardless of dominance, such that subunit vaccination targeting subdominant or recessive TCD8 specificities conferred protection against lethal poxvirus challenge. Hence, an in-depth knowledge of naturally processed T cell epitopes coupled with the identification of TCD8-based targets that protect from lethal poxviral infection are essential for rational vaccine design.
CpG-E1E2 glycosylation-mutants of Hepatitis C Virus enhance specific cellular immune response and neutralizing antibodes (113.14)
Authors: Zhang Xiao-Lian , Ren Yushan , Lin Miao , Min Yuanqin 
Published: January 2023

Abstract
Hepatitis C virus (HCV) infects 170 million people world wide, and about 80% of the infected individuals develop chronic hepatitis with a risk of progression to cirrhosis and hepatocellular carcinoma. In contrast to hepatitis A and B virus, there is no vaccine for HCV. Thus, the development of an idea vaccine against hepatitis C remains a high prority goal. N-linked glycosylation of viral proteins have been implicated in immunicity. HCV neucleocapsids consist of two highly glycosylated envelop protein E1 and E2, in which the E1 contains 5 N-linked glycans and the E2 has 11 N-glycosylation sites. These oligose chains are essential for correct folding, assemble, and antigenic variation of virus and important for inducing immune response. In this study, the effects of the N-linked glycosylation of HCV E1-E2 fusion protein, a naturally poor immunogen, on the induction of specific immune response were examined. Different E1E2 glycosylation mutants were constructed into pVAX -1 and were used as DNA vaccine. We found that some fusion E1E2 mutants and CpG-E1E2 mutants could enhance much higher specific cellular and humoral reesponses than the wild type of E1E2 in the immunized BALB/c mice. Specific mutants were determined and could be as a HCV candidcate vaccine for its higher induction of specific cellular and neutralizing antibody immune responses.
Different memory T cell phenotypes are elicited by Ad5 and rare adenoviruses (113.2)
Authors: Penaloza-MacMaster Pablo , Borducci Erica , McNally Anna , Simmons Nate , Iampietro Justin , Provine Nick , Teigler Jeff , Tan Wendy , Ahmed Rafi , Barouch Dan 
Published: January 2023

Abstract
We have compared T cell memory phenotypes and immune protection elicited by common adenoviruses serotype 5 (Ad5) versus rare adenoviruses serotypes (Ad26, Ad35, and Ad48). For these comparative studies, we immunized mice with non-replicating adenovirus vectors expressing the lymphocytic choriomeningitis virus (LCMV) GP, and challenged with chronic LCMV Cl-13. Our comparative data show that Ad5-GP generates an increased magnitude of LCMV GP-specific CD8 T cell responses (compared to that generated by rare adenoviruses). However, GP-specific CD8 T cells elicited by Ad5-GP immunization express inhibitory PD-1, and produce reduced amounts of cytokines, suggesting qualitative defects in memory CD8 T cells. This unexpected expression of PD-1 may also reflect antigen-persistence of Ad5. Moreover, Ad5-GP vaccinated mice also showed increased numbers of T regulatory cells after an anamnestic challenge with LCMV Cl-13. These data suggest that alternative serotype Ad vectors may offer substantial benefits over Ad5 as vaccine vectors in eliciting optimal T cell memory to chronic viruses such as HIV, HBV or HCV.
Multi-epitope display on virus-like particles (113.26)
Authors: Tyler Mitchell , Tumban Ebenezer , Chackerian Bryce 
Published: January 2023

Abstract
Virus-like particles (VLPs) are a flexible vaccine platform for the highly immunogenic display of peptides derived from pathogens or other vaccine targets. When designing vaccines for pathogens, it is often necessary to provoke the broadest possible immune response in order provide protection against diverse strains or serotypes. We have been interested in developing VLPs that display multiple antigenic targets. Using a recombinant PP7 bacteriophage VLP platform, we genetically inserted peptides derived from the minor capsid protein (L2) of different human papillomavirus (HPV) types and hypothesized that antibodies produced in response to vaccination will be more broadly protective than that of VLPs displaying one target only. Peptides derived from the L2 protein of multiple HPV types were displayed on the surface of recombinant PP7 bacteriophage VLPs either alone or with that of another type. Mice were immunized and the reactivity of produced antibodies was measured by through endpoint ELISA against peptides derived from an array of different HPV types as well as an in vivo heterologous HPV pseudovirus (PsV) challenge. We found that certain combinations of targets produced an antibody response with higher reactivity for heterologous HPV types than VLPs with one target alone and protected from in vivo PsV infection. Thus, displaying multiple targets on VLPs may be useful in developing broadly reactive vaccines.
Immunogenicity and specificity of Norovirus Consensus GII.4 virus-like particles in monovalent and bivalent vaccine formulations (113.27)
Authors: Parra Gabriel , Bok Karin , Taylor Ross , Haynes Joel , Sosnovtsev Stanislav , Richardson Charles , Green Kim 
Published: January 2023

Abstract
Noroviruses, a major cause of acute gastroenteritis worldwide, present antigenic diversity that must be considered for the development of an effective vaccine. In this study, we explored approaches to increase the broad reactivity of virus-like particle (VLP) norovirus vaccine candidates. The immunogenicity of a GII.4 “Consensus” VLP that was engineered from sequences of three distinct naturally-occurring GII.4 strains was examined for its ability to induce cross-reactive immune responses against different noroviruses. Rabbits immunized with GII.4 Consensus VLPs developed high serum antibody titers against VLPs derived from a number of distinct wild-type GII.4 viruses Because the sera exhibited low cross-reactivity with antigenically-distinct GI norovirus strains, we investigated the serum antibody response to a bivalent vaccine formulation containing GI.1 and GII.4 Consensus VLPs that was administered to animals under varying conditions. In these studies, the highest homologous and heterologous antibody titers to the bivalent vaccine were elicited following immunization of animals by the IM route using Alhydrogel as adjuvant. Our data indicate that the use of both genetically-engineered norovirus VLPs that incorporate relevant epitopes from multiple strains and multivalent vaccine formulations increase the breadth of the immune response to diverse variants within a genotype and, thus, prove helpful in the rational design of VLP-based vaccines against human noroviruses.