bluuebunny/crossref_metadata_2025_split huggingface dataset:
Partial organization of bovine variable-heavy chain gene locus and influence of recombination signal sequences (RSS) on various variable region gene expression (42.6)
Authors: Pasman Yfke , Kaushik Azad 
Published: January 2023

Abstract
An attempt has been made to determine the organization of variable-heavy chain gene locus in cattle, based on 91% assembled Hereford cow genome sequence, where both functional VH and pseudogenes are identified on chromosome 7 and 21. Of a total of 13 VH genes identified unambiguously, 6 VH genes are located on chromosome 21 and 7 VH genes are present on chromosome 7. Interestingly, the functional VH genes are distributed on both chromosome 7 and 21 raising questions about VDJ recombination occurring on two different chromosomes (Hosseini, A. et al. 2004. Int. Immunol.16:843) or via as yet unidentified mechanism that possibly permitted allelic exclusion. The presence of 10 DH-genes in the Hereford genome is confirmed as earlier reported by our laboratory (Koti, M. et al. 2010. Mol Immunol. 47:2119). Come cryptic recombination signal sequences (RSS) seem to exist within the VH encoding region requiring elucidation of their role in VDJ recombination. Nevertheless, V-gene expression in bovine B cells is influenced by the composition of RSSs. These findings, especially the presence of variable heavy chain locus on two chromosomes, raise fundamental questions about the phenomenon and mechanisms of allelic exclusion in bovine B cells.
A Dual-regulatory and Compartmental-specific Role for FCRL5 in Innate-like B Cell Receptor Signaling (42.8)
Authors: Davis Randall , Li Ran , Zhou Tong , Zhu Zilu 
Published: January 2023

Abstract
By virtue of their evolutionarily conserved antibody repertoires, sensitivity to T cell-independent stimuli, and rapid responsiveness, splenic marginal zone (MZ) and peritoneal cavity (PEC) B1 B lymphocytes share critical responsibility in primary humoral responses. However, despite their similar functionality they have different B cell receptor (BCR) signaling characteristics. A discrete marker of these innate-like subsets possessing tyrosine-based dual regulatory potential termed FCRL5 was investigated to explore these discrepancies. Although FCRL5 dominantly repressed BCR activation in MZ cells, it had no influence in B1 B cells. The molecular basis for its inhibition derived from SHP-1 phosphatase recruitment to its cytoplasmic ITIM. Remarkably, mutagenesis of this docking site unearthed co-activating properties for FCRL5 that were orchestrated via independent association of the Lyn kinase with its ITAM-like sequence. SHP-1 and Lyn-deficient mice validated the requirement of these elements in balancing the receptor’s unique dualistic function. These studies found that the requisite and opposing roles of SHP-1 and Lyn in modulating FCRL5’s unique binary properties directly correlated with the cellular pools of these signaling elements and consequently the disparate antigen receptor-mediated biology evident between innate-like MZ and B1 B cells.
Activated B-cells in the Granulomas of non-human primates infected with Mycobacterium tuberculosis (43.5)
Authors: Phuah Jia Yao , Mattila Joshua , Lin Philana Ling , Flynn JoAnne 
Published: January 2023

Abstract
To date, the contribution of humoral immunity, including antibodies and specific functions of B cells, on Mycobacterium tuberculosis (Mtb) infection in humans remains largely unknown. However, recent studies in mice have revealed that humoral immunity can alter Mtb infection outcomes. The cynomolgus macaque (M. fascicularis) model for Mtb infection recapitulates many aspects of human TB such as the spectrum of granulomas, variable infection outcomes, including latency and active disease, and response to treatment. As a first step to understanding the importance of humoral immunity to control of Mtb infection in primates, we characterized the B-cell and plasma cell populations in infected animals and found that B-cells are primarily present within the granuloma along with T-cells in the lymphocytic cuff. The B cells tend to be present in clusters within the granulomas are in close proximity to PNAd+ cells, have elevated HLA-DR and contain proliferating cells. Singular B-cells were also noted. Tissues containing M. tuberculosis bacilli had higher levels of Mtb specific IgG, compared to uninvolved tissue samples from the same monkeys. Plasma cells were detected within the granuloma and are functional active. This study demonstrates that B cells are present and actively secreting antibodies specific for Mtb antigens at the site of infection, in lung granulomas and thoracic lymph nodes. These antibodies likely have the capacity to modulate the host’s ability to control infection.
HMGB1 mediates Splenomegaly, Anemia and Leukocytosis in Mice Surviving Sepsis (44.7)
Authors: Valdes-Ferrer Sergio , Rosas-Ballina Mauricio , Lu Ben , Olofsson Peder , Chavan Sangeeta , Tracey Kevin 
Published: January 2023

Abstract
Severe sepsis, a syndrome complicating infection or injury, is a leading cause of death. It also has long-term deleterious effects on survivors, including high mortality and diminished quality of life. High-mobility group box 1 (HMGB1) is a critical late mediator of acute sepsis that remains elevated after other inflammatory cytokines have normalized. Here we studied the kinetics of circulating HMGB1 in mice surviving severe abdominal sepsis, and the effects of administering neutralizing anti HMGB1 mAb. In a standardized model of murine sepsis, we observed that mice surviving sepsis developed splenomegaly for at least four weeks. Splenocytes from sepsis survivors produced significantly more cytokines following exposure to TLR ligands. Circulating HMGB1 levels are significantly increased for at least 4 weeks after sepsis. Treatment with anti-HMGB1 neutralizing monoclonal antibodies effectively prevented the development of splenomegaly, leukocytosis and anemia. Chronic administration of recombinant HMGB1 to healthy mice induced significant splenomegaly and leukocytosis. Recombinant HMGB1 also sensitized splenocytes and significantly increased the release of TNF and IL-6. In conclusion, serum HMGB1 levels are persistently elevated for at least 4 weeks in mice that survive lethal sepsis. Anti-HMGB1 mAb administration reverses the development of splenomegaly, leukocytosis and anemia, which may have implications for understanding the pathogenesis of post-sepsis complications.
Generation of NK cell memory requires IL-12 and STAT4 signaling (45.3)
Authors: Madera Sharline , Sun Joseph 
Published: January 2023

Abstract
Recent studies have demonstrated that natural killer (NK) cells can become long-lived and can contribute to secondary immune responses like T and B cells; however, the precise signals that promote the generation of these long-lived memory NK cells remain unknown. Using cytokine receptor deficient mice we investigated the role of cytokine IL-12 in the activation, expansion, and generation of the virus-specific NK cell response to mouse cytomegalovirus (MCMV) infection. We demonstrate that IL-12 receptor-deficient NK cells exhibit a profound expansion defect following MCMV infection. This defect extended to the formation of long-lived memory cells, as IL-12 receptor-deficient NK cells were undetectable 2 weeks after infection whereas wildtype NK cells proliferated robustly and generated long-lived cells detectable months after MCMV infection. These findings were corroborated through the use of mice deficient in STAT4, which exhibited a defect in the expansion and formation of long-lived memory cells compared to wildtype NK cells in a competitive setting. This work demonstrates a key role for IL-12 and STAT4 during NK cell priming, and describes the requirements for generating robust virus-specific NK cell memory. Understanding the full contribution of inflammatory cytokines and the signals necessary for a long-lived NK cell response against viral infection will be of interest in the development of vaccines and therapeutics.
NKT cells enhance C5a generation by CD11b+Gr-1+ peritoneal cells via IL-10 and IFN-gamma mediated cross-talk, thereby promoting sepsis in mice (45.9)
Authors: Oh Sae Jin , Kim Ji Hyung , Chung Doo Hyun 
Published: January 2023

Abstract
Both NKT cells and complement protein C5a have been implicated in the pathogenesis of sepsis, but the issue of whether NKT cells regulate C5a generation during sepsis has been unclear. To investigate this issue, we examined immune responses in WT and NKT cell-deficient mice during cecal ligation and puncture (CLP)-induced sepsis. The levels of C5a, IFN-gamma, and IL-10 during sepsis were higher in the serum and peritoneal fluid of WT mice than in those of CD1d-/- mice. Furthermore, the mortality rate was lower in CD1d-/- mice than in WT mice. Adoptive transfer of liver mononuclear cells (LMCs) of WT mice increased mortality and C5a generation during sepsis in CD1d-/- mice, whereas transfer of IL-10- or IFN-gamma-deficient LMCs did not. In co-culture experiments, IL-10 and IFN-gamma produced by NKT cells enhanced C5a generation by CD11b+Gr-1+ peritoneal cells. Furthermore, treatment with anti-C5a antibody during sepsis decreased CD4+ T-cell apoptosis, serum D-dimer levels, and numbers of bacterial colony-forming units, but increased responsiveness against lipopolysaccharide in WT and WT-LMC-treated CD1d-/- mice, increasing survival rates. Taken together, these results suggest that CD1d-restricted NKT cells enhance C5a generation through IL-10- and IFN-gamma- mediated cross-talk with CD11b+Gr-1+ peritoneal cells, thereby aggravating CLP-induced sepsis.
Immunization with “self” peptide induces a novel phenotype of splenic dendritic cells contributing to peripheral tolerance (48.10)
Authors: Farkas Adam , Finn Olivera 
Published: January 2023

Abstract
A 100mer peptide (MUC1p) derived from the tumor associated antigen MUC1 is seen as “self” by the immune system of MUC1 Tg mice. TCR.Tg CD4+ T cells specific for MUC1 proliferate less in MUC1 Tg mice after vaccination with DC loaded with MUC1p (DC/MUC1p) relative to WT. We performed gene array on splenic RNA from WT and Tg mice vaccinated with DC/MUC1p to globally query mechanisms of peripheral tolerance to MUC1p. Expression of well-characterized pancreatic proteases (Trypsin, Elastase, CPB1) was dramatically down regulated in DC from immunized MUC1 Tg mice relative to WT, providing a signature of ensuing tolerance to self-peptide. Additionally, there was a decrease in the number of splenic DC in MUC1 Tg mice concurrent with an increase in the number of NK cells, in an IL-10 dependent fashion. IL-10 is also required to mediate down-regulation of proteases in DC linking enzyme expression with immunogenicity. Preliminary data suggest that this IL-10 is derived from increased numbers of MDSC in the spleens of immunized MUC1 Tg mice. Splenic DC from DC/MUC1p immunized MUC1 Tg mice migrate shorter distances ex vivo compared to DC recovered from WT, suggesting impaired ability to home in vivo. These DC produce less IL-12p40 and more Aldh1 compared to WT DC and express less CD40, CD86 and MHCII. Finally, DC from DC/MUC1p vaccinated MUC1 Tg mice differentiate naïve CD4+ T cells into Foxp3+ Treg in an antigen-specific manner ex vivo more effectively than DC recovered from WT mice.
Death Receptor 5 (DR5) marks the highly pathogenic interacting GM-CSF+ T helper cells and IL-23+ macrophages rendering it as an attractive therapeutic target of autoimmunity (51.4)
Authors: Li Jun , Hsu Hui-Chen , Yang PingAr , Wu Qi , Li Hao , Spalding David , Chatham W. , Kimberly Robert , Bridges S. , Mountz John 
Published: January 2023

Abstract
The highly pathogenic granulocyte-macrophage colony-stimulating factor (GM-CSF)+ IL-17+ CD4 T cells and IL-23+IRF5+M1 macrophages (MØ) exhibit a bidirectional interaction in autoimmunity. DR5 is a pro-apoptotic molecule upregulated by IRF5. DR5 deficient mice develop increased inflammation. We analyzed DR5 expression and apoptosis function of TRA-8, an anti-human DR5 antibody, in T cells and MØ in autoimmune conditions. Expression of DR5 was highly correlated with that of GM-CSF in CD4 T cells and that of IL-23 and IRF5 in MØ from synovia fluid or PBMC of rheumatoid arthritis and lupus subjects (p&lt;0.01). In vitro treatment of these samples with TRA-8 for 48 hrs resulted in 23.4% and 35.9% depletion of GM-CSF+ CD4+ T cells and IL-23+ MØ, respectively. Similar DR5 expression pattern and TRA-8 depletion effects were observed in a 3kb mouse promoter/humanized DR5 Tg mouse crossed with the SHP-1- deficient viable motheaten mice which exhibit a profound activation of inflammatory MØ and Th17 cells. Three doses of TRA-8 (0.1 mg IP, weekly) lead to a 35.7% reduction of GM-CSF+ CD4+ T cells and 45.1% reduction of IL-23+IRF5+ MØ in the draining LN, 74.4% and 94.7% decrease of Csf-2 and p19 in joints, resulting in amelioration of pneumonitis and arthritis, reduction of autoantibodies, and increase of the lifespan. Our study indicated that DR5 is highly correlated with GM-CSF in CD4+ T cells and IL-23 in MØ, and demonstrated the high therapeutic efficacy of an anti-human DR5 antibody.
Th17 cell LFA-1 engagement-mediated IL-17A and GM-CSF mRNA stabilization is HuR-dependent (54.14)
Authors: Huntenburg Sarah , Ramgolam Vinod , Yarovinsky Timur , Bender Jeffrey 
Published: January 2023

Abstract
Recent studies demonstrate that the β2 integrin LFA-1 is required for entry of encephalitogenic Th17 cells into the brain parenchyma during murine experimental autoimmune encephalomyelitis (EAE). We previously showed that LFA-1 engagement by its ligand, intracellular adhesion molecule-1 (ICAM-1), in human T cells results in translocation of the RNA-binding protein HuR and stabilization of TNFα and IFNγ mRNAs. The role of HuR in the regulation of Th17 cell-mediated inflammation is unknown. We hypothesize that LFA-1 engagement in Th17 cells leads to increased neuropathology-relevant cytokine expression at inflammatory sites through HuR-mediated mRNA stabilization. LFA-1 engagement led to HuR nuclear-to-cytoplasmic translocation and stabilization of both IL-17A and GM-CSF transcripts in mouse EL4 T cells, in vitro differentiated mouse Th17 cells and human primary T cells. EL4 and human primary T cell adhesion to ICAM-1 led to increased levels of IL-17A and GM-CSF mRNA in HuR immunoprecipitates. LFA-1 engagement-stimulated IL-17A and GM-CSF mRNA stabilization was lost in HuR-deficient Th17 cells generated from RORcγt-Cre/HuRflox/flox mice. We conclude that LFA-1 engagement induces HuR-dependent stabilization of the otherwise labile GM-CSF and IL-17A transcripts in Th17 cells, and that this may be highly relevant in Th17 cell-induced CNS pathology. Future experiments will focus on the role of Th17 cell HuR in the EAE model.
Rheumatoid arthritis synovial tissue fibroblast Mucin 3 expression and function (54.27)
Authors: Volin Michael , Zanotti Brian , Klosowska Karolina , Smith Kelley , Birly Jennifer , Rodriguez Manuel , Woods James 
Published: January 2023

Abstract
Mucin 3 (MUC3) is elevated in rheumatoid arthritis (RA) synovial tissues (STs). We hypothesized that MUC3 is expressed by RA synovial fibroblasts, that it is cytokine inducible and chemotactic for RA synovial fibroblasts. To test this, RA synovial fibroblast MUC3 expression was determined by immunohistochemistry, flow cytometry and Western blot analysis. Levels of MUC3 from arthritic synovial fluids (SFs) were measured by a novel MUC3 ELISA. RA synovial fibroblast chemotaxis to RA SFs was assessed using a modified Boyden chamber assay. RA ST cells stained for both MUC3 and vimentin indicating that RA synovial fibroblasts express MUC3. Flow cytometry experiments showed that most of the RA synovial fibroblast MUC3 is cytoplasmic. Cytokine stimulation of these cells significantly increased MUC3 production (P&lt;0.05). RA SFs contain on average 8.7μg/ml (n=8) of MUC3. Immunodepletion of MUC3 from RA SFs resulted in significantly reduced RA synovial fibroblast chemotaxis (P&lt;0.05, n=5 RA SFs and n=8 RA synovial fibroblasts). These results show that MUC3 is produced by synovial fibroblasts in RA STs, that MUC3 is inducible by cytokine stimulation and that RA SF induced synovial fibroblast chemotaxis can be inhibited by depleting MUC3. Thus, MUC3 synovial fibroblast production may be induced by the inflammatory milieu of the RA joint and may drive synovial fibroblast chemotaxis within the RA ST potentially aiding pannus formation and cartilage destruction.
Modulation Of IL-17A And TL1A Largely Abrogates House Dust Mite-Induced Lung Inflammation In A Murine Model Of Allergic Airway Disease (54.5)
Authors: Hubeau Cedric , Kubera John , Hammerman Katherine , Wright Jill , Denz Michelle , Juang Yuang-Taung , Williams Cara 
Published: January 2023

Abstract
Recent studies suggest a role for Th17 responses in the increased airway neutrophilia associated with severe asthma. House dust mite (HDM) is a natural allergen to which asthmatics are often sensitized. Mice repeatedly challenged with HDM extract developed robust airway neutrophilia rapidly evolving into asthma-like disease with increased numbers of eosinophils and lymphocytes in bronchoalveolar lavages (BAL) as well as inflammatory infiltrates, vascular/muscular hypertrophy, interstitial fibrosis, epithelial hyperplasia and mucus accumulation in lung tissues. RNA and protein screening revealed a robust Th17 component post-HDM exposure. We thus evaluated whether IL-17A deficiency could modulate HDM-induced allergic airway disease. Airway neutrophilia was indeed abrogated in IL-17A deficient mice weekly challenged with HDM (acute model), however total BAL cellularity and lung mechanics remained comparable to those of HDM-challenged WT mice. In contrast, IL-17A deficient mice daily exposed to HDM (chronic model) had decreased BAL cellularity associated with reduced numbers of BAL macrophages, neutrophils, eosinophils and lymphocytes. Interestingly antibody neutralization of TL1A, a member of the TNF superfamily known to promote Th2 and Th17 responses, reduced BAL cellularity to baseline levels in HDM-challenged WT mice. Our results thus indicate that targeting Th17 responses can alleviate HDM-induced airway neutrophilia, and can also broadly modulate allergic airway disease.
IRAK4 kinase-deficient mice exhibit increased susceptibility to S. pneumoniae that is overcome by poly IC:LC treatment (55.23)
Authors: Pennini Meghan , Salazar Andres , Vogel Stefanie 
Published: January 2023

Abstract
Interleukin-1 receptor-associated kinase 4 (IRAK4) is a critical component of Toll-like receptor signaling, and patients with IRAK4 mutations are extremely susceptible to recurrent bacterial infections. In these studies, we used mice homozygous for a mutant IRAK4 that lacks kinase activity (IRAK4KDKI) to address the role of IRAK4 in response to TLR agonists and infection with whole bacteria. IRAK4KDKI macrophages have a diminished response to the TLR4 agonist LPS, little to no response to the TLR2 agonist Pam3Cys, but normal response to the TLR3 agonist poly I:C when compared to wild-type (WT) macrophages as measured by cytokine mRNA expression and MAPK activation. Thus, diminished responses of IRAK4KDKI cells are specific to TLR pathways that are MyD88-dependent. Importantly, we have identified two proteins downstream of MAPK signaling, MNK1 and MSK1, whose phosphorylation is deficient in IRAK4KDKI macrophages when compared to WT. These data suggest a role for IRAK4 in propagating a TLR signal beyond initial phosphorylation of p38 and ERK. Additionally, IRAK4KDKI mice were more susceptible to infection with S. pneumoniae, demonstrating the usefulness of this murine model for the exploration of strategies to overcome IRAK4 deficiencies. Co-treatment of infected mice with Poly IC:LC, a TLR3 agonist, significantly improved the survival rate of both WT and IRAK4KDKI mice providing a potential treatment strategy in both normal and immunocompromised patients.
Role of IL-17A in airway tolerance in a murine model of asthma (57.14)
Authors: Graser Anna , Zhivkova Nataliya , Finotto Susetta 
Published: January 2023

Abstract
Allergic asthma is a world-wide increasing disease. The mechanism controlling this disease is not yet completely understood. In the attempt to design vaccination strategies against it, we previously established a model of airway tolerance. Airway tolerance is characterized by decreased pro-inflammatory immune responses. In this model we described in wild type as well as in T-bet deficient mice a down-regulation of both Th2 and Th17 cytokines indicating a T-bet independent mechanism in this model. In this study we found that this feature is accompanied by induction of the suppressive cytokine IL-10 in the airway tolerance model only in the T-bet deficient mice. Because IL-17A is a T cell-derived pro-inflammatory cytokine playing a critical role in immune and inflammatory responses we asked about the role of IL-17A in the airway tolerance model by using IL-17A deficient and wild type mice. In our experiments we found that IL-17A deficient mice had decreased airway hyper-responsiveness after methacholin challenge in the airway tolerance model as compared to allergic asthma model. Moreover we found an increase of IL-10 protein release in the airways of IL-17A deficient mice as compared to the wild type littermates in this model. These results indicate a suppressive function of IL-17A on IL-10 in the presence of T-bet in the lung that should be further investigated.
In utero exposure to EPFR-containing PM inhibits pulmonary Th1 maturation and enhances adult asthma development in mice (57.15)
Authors: Cormier Stephania , Wang Pingli , You Dahui 
Published: January 2023

Abstract
Environmentally persistent free radicals (EPFRs), with half-lives on the order of weeks have been detected in association with airborne particulate matter (PM) and early exposure to environmental hazards, such as PM, has been associated with an increased incidence of asthma in childhood. We hypothesized that in utero exposure to EPFR-containing PM disrupts immune development resulting in aberrant immune responses and enhancing susceptibility to asthma. To test this hypothesis, we exposed pregnant mice to ultrafine particles containing persistent free radicals (EPFR-UFP) and determined postnatal immune development and response to allergen sensitization and challenge. Intrauterine exposure to EPFR-UFP inhibited pulmonary T helper (Th1/Th2/Th17) and T regulatory (Treg) responses in early life (&lt;1 week of age). Th2 and Treg responses recovered to normal levels; whereas Th1 responses remained attenuated at 6 weeks of age. In the asthma model, intrauterine EPFR-UFP-exposed mice exhibited higher airway hyperresponsiveness and pulmonary eosinophilia. In addition, levels of IL-4 and IL-5 (Th2 cytokines) in the bronchoalveolar lavage fluid of these mice were significantly higher than that of non-exposed mice that were sensitized and challenged with ovalbumin. Our data suggest that in utero exposure to EPFR-UFP enhances adult asthma development in mice, which might be related to the inhibition of pulmonary Th1 maturation.