bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of glucocorticoid receptor in B lymphocytes (111.3)
Authors: Gruver-Yates Amanda , Cidlowski John 
Published: January 2023

Abstract
Glucocorticoids play an important role in the resolution of inflammation. They are routinely used to treat allergies, asthma, sepsis, and autoimmune diseases. In addition to immune suppression, glucocorticoids can negatively impact the development and function of the immune system by inducing apoptosis, an effect well-studied in mature and developing T cells. Little is known however about glucocorticoid function specifically in B lymphocytes. We have shown that glucocorticoid receptor (GR) is present in enriched B cell populations isolated from mouse spleen. GR protein was present in all B cell (B220+) developmental subsets (Mature IgM+IgD+, Immature IgM+IgD-, and Pro/Pre IgM-IgD-) isolated from spleen. Intracellular staining for GR analyzed with flow cytometry also confirmed the presence of GR in all B cell subsets from cells isolated both from mouse spleen and bone marrow. Mean fluorescence intensity data indicated that Pro/Pre B lymphocytes had more intense staining of GR than either mature or immature populations. Ex vivo cell culture of murine splenocytes indicated that dexamethasone, a synthetic and potent glucocorticoid, reduced cell viability in culture in all B cell developmental subsets. In vivo administration of dexamethasone reduced total cell numbers of all B cell subsets present in the mouse spleen within 24 hours. These data suggest that B cells glucocorticoid signaling through GR on B cells have an important role in B cell function.
BTB-POZ transcription factors recruit the E3 ubiquitin ligase Cullin 3 to initiate lymphoid effector programs (111.38)
Authors: Mathew Rebecca , Sieler Michael , Scanlon Seth , Constantinides Michael , Bertozzi-Villa Clara , Singer Jeffrey , Bendelac Albert 
Published: January 2023

Abstract
The differentiation of several T and B cell effector programs in the immune system is directed by signature transcription factors that induce rapid epigenetic remodeling. We report that PLZF, the BTB-POZ transcription factor directing the innate-like effector program of NKT thymocytes was prominently associated with cullin 3 (Cul3), an E3 ubiquitin ligase previously shown to use BTB domain-containing proteins as adaptors for substrate binding. PLZF induced the transport of Cul3 from the cytosol to the nucleus where the two proteins were associated within a chromatin associated/modifier protein complex. Furthermore, PLZF expression in thymocytes resulted in selective changes of ubiquitination of multiple components of this complex. Cul3 was also found associated with another BTB-POZ transcription factor, Bcl6, which directs the B cell germinal center program. Conditional deletion in mice demonstrated an absolute, cell-intrinsic requirement of Cul3 for the development of NKT cells and germinal center B cell responses. We conclude that distinct lineage-specific BTB-POZ transcription factors recruit Cul3 to alter the ubiquitination pattern of their associated chromatin associated/modifier complex. We propose that this novel function is essential to direct the differentiation of several T and B lymphocyte effector programs, and may also be involved in the oncogenic role of PLZF and Bcl6 in leukemias and lymphomas.
Cellular and molecular profiling of CD8 T cell differentiation with respect to positive and negative selection in vitro reveals a role for Notch signaling (111.45)
Authors: Dervovic Dzana , Liang Haydn , Cannons Jennifer , Elford Alisha , Ohashi Pamela , Schwartzberg Pamela , Zuniga Pflucker Juan Carlos 
Published: January 2023

Abstract
Differentiation of functionally mature CD8 single positive (SP) T cells is predicated by the ability of lymphocyte progenitors to integrate multiple signaling cues provided by the thymic microenvironment. In thymus, signaling through Notch receptors directs T cell progenitors to commit to the T cell lineage, undergo early T cell differentiation, and progress to the CD4+CD8+ (DP) stage of T cell development. However, it is still unclear whether Notch signals are prerequisite for differentiation or commitment of DP cells to CD8 SP stage of T cell development. Here, we demonstrate that ectopic expression of the Notch-ligand by OP9 stromal cells allows for efficient differentiation of conventional effector CD8 T cells from SAP-/- bone marrow (BM)-derived hematopoietic stem cells (HSCs). We also prove that OP9-DL1 stromal cells restrict differentiation of innate CD8 T cells but allow for differentiation of IL17-producing CD8 T cells from BM-HSCs isolated from Itk-/-Rlk-/- (DKO) mice. We further reveal that the process of positive and negative selection in vitro is constrained by peptide-MHC (pMHC) class I expressed by the OP9 stromal cells. Finally, using a T cell receptor (TCR) transgenic model we disclose that the commitment of DP precursors to the CD8 T cell lineage is facilitated by Notch signaling. Our findings further establish the requirement for Notch receptor-ligand interactions throughout intrathymic T cell differentiation.
The role of IRF4 in regulating effector CD8+ T cell development via repression of EOMES expression. (111.51)
Authors: Nayar Ribhu , Enos Megan , Prince Amanda , Shin Hyun Mu , Hemmers Saskia , Klein Ulk , Rudensky Alexander , Berg Leslie 
Published: January 2023

Abstract
CD8+ T cell development in the thymus generates a predominant population of conventional naïve cells, along with minor populations of ‘innate’ T cells that resemble memory cells. Recent studies analyzing a variety of knockout or knock-in mice have indicated that impairments in the TCR signaling pathway produce increased numbers of innate CD8+ T cells, characterized by their high expression of CD44, CD122, CXCR3, and the transcription factor Eomesodermin. One component of this altered development is a non-CD8+ T cell-intrinsic role for IL-4. To determine whether reduced TCR signaling within the CD8+ T cells might also contribute to this pathway, we investigated the role of the transcription factor IRF4. IRF4 is upregulated following TCR stimulation in wild-type T cells; however, this upregulation is impaired in Itk-/- T cells, which have reduced responses to TCR signaling. Further, analysis of IRF4-deficient CD8+ thymocytes showed normal development of thymic CD8+ T cells. Interestingly, IRF4-deficient peripheral CD8+ T cells acquired a memory phenotype and expressed the transcription factor Eomesodermin. We also show that activation of naïve IRF4-deficient CD8+ T cells leads to rapid and robust expression of Eomesodermin. Together, these data indicate that IRF4 upregulation following CD8+ T cell activation normally suppresses Eomesodermin expression, thereby regulating the differentiation pathway of CD8+ effector T cells.
PAD4 regulates proliferation of hematopoietic multipotent progenitors by directly controlling c-Myc expression. (111.7)
Authors: Nakashima Katsuhiko , Arai Satoko , Miyazaki Toru 
Published: January 2023

Abstract
Peptidylarginine deiminase 4 (PAD4) functions as a transcriptional co-reporessor by catalyzing the conversion of arginine to citrulline residues of histone H3. Although the high expression of PAD4 in the bone marrow may suggest its involvement in the hematopoiesis, its precise contribution remains unclear. Here we found that PAD4 is highly expressed in the hematopoietic stem and multipotent cells (Lineage-Sca1+c-kit+; termed LSK cells). Microarray and RT-qPCR analysis showed that the mRNA level of c-myc in PAD4-deficient (PADI4-/-) LSK cells was increased compared to PADI4+/+ LSK cells. Chromatin immunoprecipitation assay showed that in LSK cells, PAD4 targeted to the upstream of c-myc gene, suggesting that PAD4 might catalyze the citrullination and reduce the arginine methylation in histone H3. Furthermore, PAD4 was associated with the transcription factor LEF1 as well as histone deacetylase 1 (HDAC1). These results indicated that PAD4 suppressed c-Myc expression by histone citrullination acting as a suppressor complex with LEF1 and HDAC1 in LSK cells. Supporting these findings, PADI4-/- LSK cells showed an advanced proliferation in a cell-autonomous fashion, resulting in increase in number compared to PADI4+/+ LSK cells. Together, our results strongly suggest that PAD4 regulates the proliferation of LSK cells via controlling c-Myc expression, implying that PAD4 may be involved in pathogenesis of some myeloproliferative diseases.
MAP Kinase Phosphatase 5 Protects Against Sepsis-induced Acute Lung Injury (112.10)
Authors: Qian Feng , Deng Jing , Gantner Benjamin , Dong Chen , Christman John , Ye Richard 
Published: January 2023

Abstract
Mitogen-activated protein kinases (MAPKs) play a critical role in inflammation. While activation of MAPK in inflammatory cells has been studied extensively, much less is known about the inactivation of these kinases. MAPK phosphatase 5 (MKP5) is a member of the dual specificity phosphatase family that dephosphorylates activated MAPKs. Here we report that MKP5 protects sepsis-induced acute lung injury. Mice lacking MKP5 displayed severe lung tissue damage following LPS challenge, characterized with increased neutrophil infiltration and edema compared to wild type controls. In response to LPS, MKP5-deficient macrophages produced significantly more inflammatory factors including inflammatory cytokines, nitric oxide (NO), and superoxide. Phosphorylation of p38 MAPK, JNK and ERK were enhanced in MKP5 deficient macrophages upon LPS stimulation. Adoptive transfer of MKP5-deficient macrophages led to more severe lung inflammation than transfer of WT macrophages, suggesting that MKP5-deficient macrophages directly contribute to acute lung injury. In addition, mice lacking MKP5 showed higher mortality rate than WT mice following cecal ligation and puncture (CLP), although MKP5 deficiency enhanced bacterial clearance by macrophages. Taken together, these results suggest that MKP5 is crucial to homeostatic regulation of MAPK activation as well as the balance of antimicrobial and inflammatory responses.
Coordinated influence of β-defensins on innate immunity against bacterial infection of the airway (112.12)
Authors: Ryan Lisa , Wu Jichuan , Yim Sunghan , Shah Helly , McMahon Laura , Minhas Abeer , Diamond Gill 
Published: January 2023

Abstract
β-defensins act as antimicrobial agents and as immunomodulators. Human β-defensin (BD) peptides 1, 2 and 3 are differentially modulated in primary tracheal epithelial cells (TEC) by viruses, bacteria, and proinflammatory cytokines. We hypothesized that hBD-1 and virulence factors would influence induction of hBD-2 and hBD-3 during lung bacterial infections. Since BD genes exist in close proximity on chromosome 8, making it impossible to delete more than one BD gene, a model was devised to study their coordination. Since mouse orthologs exist, (hBD-1=mBD-1; hBD-2=mBD-3; hBD-3=mBD14), we used mBD-1(-/-) (KO) mice and the inhibitory capability of the type III secretion system of Bordetella bronchiseptica on NF-κB to study effects of mBD-1 deletion on mBD-3 gene expression using wild-type RB50 and mutant WD3 strains lacking this system. In C57BL/6 wild-type (WT) mice, mBD-3, mBD-14 and kerotinocyte-derived chemokine (KC) mRNA levels were suppressed by RB50 compared with WD3 induction in lung 24 hr after infection, whereas mBD-1 levels were unaffected. Deletion of mBD-1 significantly blunted WD3 induction of mBD-3 and mBD-14, but not KC. Bacteria numbers increased, along with decreased neutrophils in the trachea, when KO mice were infected with RB50 compared with WT mice. In TEC, WD3 induced mBD-3 in WT but not in KO mice, yet with cultured KO TEC, mBD-14 and KC were unaffected. The results illustrate the coordination of BD in TEC and in lung to influence bacterial infection.
Specific modulation of host intestinal epithelial NF-kB signaling and inflammatory response by E. coli O157:H7 effector proteins (112.20)
Authors: Wan Fengyi 
Published: January 2023

Abstract
Intestinal epithelial NF-kB signal pathways are crucial for host response to pathogen infections by orchestrating the innate and adaptive immune responses. Many successful pathogens have acquired sophisticated mechanisms to regulate host NF-kB signaling in order to favor the continued pathogen survival. A repertoire of novel virulence proteins, namely Nle effectors, was identified in the foodborne pathogen Escherichia coli O157:H7, and most of them appeared to subvert host NF-kB activation. However, the mechanism(s) through which the effectors proteins manipulate NF-kB signaling and inflammatory response in host is still largely unknown. We discovered that the NleH1 and NleH2 effectors from E. coli O157:H7 subvert host intestinal epithelial NF-kB signaling through their interactions with ribosomal protein S3 (RPS3), a non-Rel subunit of NF-kB that confers certain transcriptional specificity. NleH1 selectively attenuates, whereas NleH2 facilitates, RPS3-dependent proinflammatory cytokine genes, although both effectors do not impair p65 nuclear accumulation. We further demonstrated that NleH1 and NleH2 execute distinct function in regulating the Inhibitor of kB kinase 2 (IKK2)-mediated RPS3 phosphorylation and subsequent nuclear translocation. Thus, our results reveal a novel mechanism on the specific modulation of host NF-kB activation and inflammatory response by NleH effectors, and provide new molecular insights into the complex pathogenic mechanisms of E. coli O157:H7.
mTOR modulates host immune responses to Francisella tularensis LVS infection (112.7)
Authors: Edwards Michael , Zhang Ping , Michalek Suzanne , Katz Jannet 
Published: January 2023

Abstract
Francisella tularensis (Ft) is the cause of tularemia, a disease with a high morbidity and mortality rate. The Ft live vaccine strain (LVS) has been used to study disease pathogenesis since infection of mice with LVS resembles tularemia in humans. It has been reported that Ft LVS is a TLR2 agonist and involves PI3K/Akt signaling. However, the involvement of mTOR, a downstream molecule of the PI3K/Akt pathway, has not been determined in Ft LVS infection, nor has the interplay between TLR2 and the mTOR cascade. In the present study, we provide evidence that mTOR exerts a differential regulation on both cytokine mRNA and secreted protein in murine peritoneal macrophages stimulated with Ft LVS. IL-6, IL-10 and IFN-β mRNA production increased with inhibition of the mTOR pathway, while levels of IFN-γ mRNA were independent of mTOR signaling. Secreted IL-6 and IFN-γ but not IFN-β and TNFα was dependent of mTOR regulation. Furthermore, mTOR signaling was critical for the DNA binding activity of CREB, but not of NF-κB p65. Upregulation of the interferon regulatory factors (IRF)-1, IRF-3 and IRF-5 in nuclear fractions of cell lysates was observed upon inhibition of mTOR, and correlated with increased levels of the pro-apoptotic proteins Bax, caspase 3 and caspase 8, indicating mTOR modulation of apoptosis via IRF downreguation. Overall, these studies identify mTOR as a critical regulator of the host immune response to Ft LVS infection.
Anti-vector immunity following vaccination with Venezuelan equine encephalitis replicon filovirus vaccine (113.1)
Authors: Zak Samantha , Kuehne Ana , Barth James , Herbert Andrew , Muhammad Majidat , Ortiz Ramon , Prugar Laura , Hannah Brian , Bakken Russell , Pratt William , Dye John 
Published: January 2023

Abstract
The Venezuelan equine encephalitis replicon (VRP) is the lead candidate for development as the DoD filovirus vaccine platform. One concern regarding this vaccine candidate focuses on the need to vaccinate individuals that have been pre-exposed to the VRP through previous vaccinations. To address this concern, cohorts of nonhuman primates (NHPs) were vaccinated with VRP expressing alphavirus proteins and VRP expressing Ebola virus Zaire (EBOV) glycoprotein either concurrently or sequentially. These NHPs were challenged with either 1000pfu of EBOV or 10^8 pfu of Venezuelan equine encephalitis virus (VEEV). Animals that were vaccinated with both vaccinations independent of sequence or co-administration were protected from each challenge. This data indicates that anti-vector immunity is not a concern with the VRP vaccine candidate as the product transitions into phase 1 clinical trials.
Alum adjuvant rescues HPAI vaccine immunogenicity and survival in BALB/c mice following H5N1 infection (113.30)
Authors: Vaughan Sarah , Stout-Delgado Heather , Pollock Zemmie , Plourde Jennifer , Pyles John , Harrod Kevin 
Published: January 2023

Abstract
Highly pathogenic avian influenza (HPAI) is an emerging infectious virus with an approximate 60% fatality rate in humans. In the United States, a vaccine for H5N1 has been developed and stockpiled in case of a pandemic. This vaccine demonstrated low immunogenicity in human clinical trials with only 22% of recipients developing protective antibody titers. Previous studies using a lethal challenge ferret model have shown that supplementing the H5N1 vaccine with an alum adjuvant can improve immunogenicity and survival. We developed a lethal challenge mouse model of H5N1 vaccination to examine differences in innate immune responses to the vaccine alone or in the presence of an alum adjuvant. Mice treated with the adjuvanted vaccine showed significantly lower weight loss and higher survival than mice treated with the nonadjuvanted vaccine. Further, mice treated with adjuvanted vaccine had higher neutralizing antibody titers when compared to mice that received the nonadjuvanted vaccine. In vitro, dendritic cells (DCs) cultured with the adjuvanted vaccine had increased expression of the activation markers CD40 and CD86 compared to DCs treated with the nonadjuvanted vaccine. Interestingly, neither treatment induced activation of Toll-like receptors or the production of cytokines IL-6, IL-12, or TNF-α. Taken together, our results suggest that the alum adjuvant rescues immunogenicity and improves survival in the host.
Protective efficacy of CD4 memory T cells depends on route of administration of inactivated influenza vaccine (113.31)
Authors: Jelley-Gibbs Dawn , Winfrey Carolyn , Eaton Sheri , Haynes Laura 
Published: January 2023

Abstract
Studies aimed at determining the impact route of influenza vaccination has on vaccine efficacy revealed reduced protection against influenza virus challenge afforded by intranasal delivery compared to subcutaneous delivery of inactivated H1N1 influenza A virus. Both vaccine routes induced similar anti-viral antibody titers, similar numbers of anti-viral CD4 T cells, and undetectable anti-viral CD8 T cell responses. Further analysis of vaccine-induced CD4 memory T cell responses revealed that CD4 memory T cells primed in the lung draining lymph nodes were defective in recruiting anti-viral CD8 effectors to the lungs during challenge infections compared to the recruitment of CD8 effectors to the lungs by peripherally primed CD4 memory T cells. Since the influx of anti-viral CD8 effector T cells to the airways during influenza virus infection is a key event for virus control and elimination, we assayed for the expression of type-1 chemoattractants in the respiratory tract of vaccinated mice following challenge infection and discovered reduced lung expression of chemoattractants in the presence of intranasally primed CD4 memory T cells. We also determined that the presence of intranasally primed CD4 memory T cells reduced the ability of pulmonary antigen presenting cells to stimulate CD8 effectors directly ex vivo. We are currently assessing whether route of influenza vaccination also impacts susceptibility to secondary bacterial pneumonia following influenza challenge.
Harnessing Sendai virus to induce mucosal immunity (113.32)
Authors: Le Thuc-vy , Garcin Dominique , Compans Richard 
Published: January 2023

Abstract
The human mucosal surface covers greater than 400 m2 and is a target for infection and replication of pathogens such as influenza virus and HIV. Induction of specific immunity at the site of pathogen entry is therefore critical for the prevention of infection. We generated an attenuated recombinant Sendai virus (GP42-H1) in which the hemmagglutinin (HA) gene of A/Puerto Rico/8/1934 (PR/8) influenza virus was introduced into the genomic RNA. Intranasal (i.n.) instillation of mice with 1,000 pfu of the vector did not lead to weight loss. Expression of recombinant protein was observed in vivo, within 4 days post immunization. Heterologous expression of the recombinant gene contributed to high titer of HA-specific IgG and IgA antibodies in blood, saliva, fecal pellet extracts and bronchoalveolar lavage fluid. These antibodies inhibited hemagglutination of chicken erythrocytes and neutralized influenza virus infection of Madin-Darby Canine Kidney cells in vitro. Following challenge with 5LD50 of mouse adapted PR/8 influenza virus, mice that were immunized with 1,000 pfu of the GP42-H1 vector were completely protected from the influenza infection as opposed to mice that were immunized with a vector control that expresses green fluorescent protein. These data demonstrate that i.n. immunization with the GP42-H1 vector is competent to induce immunity at mucosal tissues and immunization with the vector promotes specific protection from lethal challenge in mice.
Role of inflammasome in HSP-mediated innate immunity (114.5)
Authors: Pawaria Sudesh 
Published: January 2023

Abstract
The immunogenic heat shock proteins gp96, hsp70 and calreticulin stimulate antigen presenting cells to provide co-stimulation in the form of cytokines and co-stimulatory molecules. The unique pattern of co-stimulation is dependent on the type of antigen presenting cell and the HSP that is used as the stimulant. The pattern of co-stimulation also determines the T helper response that is primed. A cytokine that is routinely released by HSP-stimulated APCs is IL-1beta. Here we investigate the role of (components of) the inflammasome pathway in HSP-mediated IL-1beta release. Our results show that in response to HSP stimulation and in a CD91-dependent manner, first, antigen presenting cells secrete robust amounts of IL-1β comparable to LPS stimulation, second, the HSP receptor CD91 is phosphorylated, and third, downstream p38 and NF-kB are activated. The source of the second signal which allows for the conversion of the pro-IL1beta to the mature form in this system is being investigated.
IP4 regulates FasL-mediated T lymphocyte death via inhibition of Orai1 (115.1)
Authors: Miller Andrew , Dahlberg Carol , Parker Albert , Schmedt Christian , Beisner Daniel , Gosling Martin 
Published: January 2023

Abstract
Antigen receptor-mediated production of Ins(1,4,5)P3 induces the release of Ca2+ from intracellular stores, resulting in the opening of store-operated Ca2+ (SOC) channels. We previously reported a mouse line deficient in Inositol(1,4,5)P3-3 kinase B (ITPKb), which converts Ins(1,4,5)P3 (IP3) to Ins(1,3,4,5)P4 (IP4). Mice lacking ITPKb exhibit a complete block in T cell positive selection and possess impaired B cell development and activation. Interestingly, ITPKb-/- cells exhibit enhanced Ag receptor-induced SOC signaling, which can be rescued by adding cell-permeable IP4 to ITPKb-/- cells, suggesting that IP4 serves to inhibit SOC channels. Due to developmental blocks, the role of ITPKb in mature lymphocyte function is unknown. Using the ER-Cre-loxP system, tamoxifen-induced deletion of ITPKb (ITPKbfl/fl) demonstrates a crucial role for ITPKb in mature T cell function. While ITPKbfl/fl mice have normal T-independent Ab responses, T-dependent Ab responses are completely abolished. In vitro, mature ITPKbfl/fl T cells exhibit enhanced SOC entry and reduced proliferative responses to TCR signals. Furthermore, we found that ITPKbfl/fl T cells die rapidly after activation, due in part to enhanced FasL upregulation. Using a HEK293 cell line stably expressing Stim1 and Orai1, we demonstrate that IP4 is an inhibitor of open-state Orai1 channels. These data identify ITPKb and IP4 as essential mediators of lymphocyte development and T cell activation by regulating SOC entry via Orai1.