bluuebunny/crossref_metadata_2025_split huggingface dataset:
Suppression of experimental epidermolysis bullosa acquisita (EBA) by an IgG monoclonal antibody (mAb) to collagen type VII (COL7)
Authors: Samavedam Unni Krishna SRL , Potter Crystal , Angerman Elizabeth , Kasprick Anika , Bieber Katja , Ludwig Ralf , Finkelman Fred D 
Published: January 2023

Abstract
EBA is a chronic inflammatory, mucocutaneous blistering skin diseases caused by autoantibodies against the structural skin protein, COL7. EBA induction requires autoantibody binding as well as complement- and Fcγ receptor (FcγR)-dependent inflammation. Using mouse models of EBA, we evaluated: (1) in IgG1-deficient mice, whether preventing the production of IgG1 anti-COL7 Ab (IgG1 has limited ability to activate complement or FcγRs), influences EBA induced by COL7/adjuvant immunization (active model); and (2) whether a mAb of the pro-inflammatory IgG2c isotype can induce EBA or influence passive EBA induction by a rabbit IgG anti-mouse COL7 or active EBA induction. Disease in the active EBA model develops in half the time and is more severe in genetically susceptible IgG1-deficient mice than in wild-type mice. Blocking IFN-γ, which contributes to IgG2c induction, has little or no effect. 16A1C8, an IgG2c monoclonal anti-COL7 mAb, does not induce EBA, even in hypersusceptible FcγRIIB-deficient mice. Surprisingly, 16A1C8 suppresses EBA induction by rabbit anti-COL7 Ab, and EBA progression in COL7-immunized mice, even though IgG2c is a pro-inflammatory isotype, 16A1C8 does not compete with our rabbit Ab for binding to COL7, and 16A1C8 has little or no effect on neutrophil infiltration or C3 deposition at the dermal-epidermal junction. Thus; (1) autoantibodies that have limited ability to induce pro-inflammatory effector mechanisms can suppress EBA development and severity; and (2) some anti-COL7 mAbs, even those of a pro-inflammatory isotype, may be useful for preventing or even treating EBA.
H5P as an active learning tool builds content-based critical thinking skills in Biological Sciences and student confidence in problem-based learning
Authors: Carr William H , Barry Awa 
Published: January 2023

Abstract
Recent studies have demonstrated that active learning or learning by doing has a more significant positive impact on learning outcomes than reading or watching videos. However the availability of free and open source tools for creating these active learning exercises has been limited by the requirement of computer programing skills. With the development of H5P technology, an open source, on-line tool (www.h5p.org), faculty and even undergraduate students can create interactive exercises that facilitate the learning process and promote formative learning. H5P is a plugin for existing publishing systems that enables the system to create interactive formative assessments, such as multiple-choice quizzes that provide feedback in real-time on responses. Using this technology I created an active learning exercise that was implemented in an undergraduate General Biology I course for Biology majors at Medgar Evers College, CUNY, in Fall 2019. In this exercise students designed interactive multiple choice questions at both low and high levels of Modified Bloom’s Taxonomy. Based on surveys of their subjective impressions, a majority (73%) of the students reported that their confidence increased in explaining course-content (n=44). Content knowledge was also assessed before and after implementing this exercise This exercise was also implemented in an undergraduate Immunology course (n=11) on the topic of the regulation of V(D)J recombination. Here, I present how to introduce and implement this exercise among undergraduate students who are Biology Majors. The outcomes from this work suggest that this platform is an effective and versatile tool for building student confidence in explaining scientific content in the biological sciences.
Elucidating the Role of NEMO and SpiC in DNA Double Strand Break Induced Inhibition of V(D)J Recombination
Authors: Glynn Rebecca , Bassing Craig 
Published: January 2023

Abstract
V(D)J recombination is regulated such that most lymphocytes assemble and express an antigen receptor from only one allele (allelic exclusion). In pre-B cells, RAG-induced DNA double strand breaks (DSBs) at Igκ loci signal via ATM to rapidly repress Rag1/2 transcription, inhibit accessibility of Igk loci, and limit V recombination. We hypothesize that RAG DSB-induced repression of Rag1/2 is critical to transiently limit V recombination, ensure allelic exclusion, and inhibit oncogenic Ig translocations. Fittingly, Atm−/− mice have higher frequencies of developing B cells with RAG DSBs at both Igh and Igk loci and of mature B cells with bi-allelic expression of these genes. Yet, given the multifunctional roles of ATM in the DSB response, Atm−/− phenotypes cannot be directly attributed to Rag1/2 repression. To test our hypothesis, we study mouse models lacking downstream effectors of ATM: i) SpiC, which suppresses Igk accessibility in response to RAG DSBs, and ii) NFκB essential modulator (NEMO), which represses Rag1/2 in response to genotoxic DSBs. I show here that the frequency of surface bi-allelic Igκ expression is normal on SpiC−/− B cells and increased on Nemo−/− B cells. I also show that Nemo−/− pre-B cells exhibit impaired Rag1/2 repression in response to RAG DSBs and concomitant increased RAG cleavage at Igk. Our data support that RAG DSB-induced repression of Rag1/2 transiently inhibits additional Igk rearrangements to enforce Igκ allelic exclusion independent of ATM-mediated DSB repair. I am using Nemo−/− mouse models to determine if DSB-induced Rag1/2 repression requires Nemo in pro-B cells, and if this mechanism is critical to suppress oncogenic Ig translocations independent of defects in DSB repair/checkpoint activation.
E protein support of Zeb2 expression regulates development of multiple hematopoietic lineages
Authors: Huang Xiao , Ferris Stephen , Satpathy Ansuman , Murphy Theresa L , Murphy Kenneth M 
Published: January 2023

Abstract
Zeb2 is an E-box binding transcriptional repressor that were first discovered to be important for epithelial to mesenchymal transition. We recently discovered a genetic circuit involving Zeb2, Nfil3, Id2 that governs dendritic cell (DC) specification in the multipotent common dendritic cell progenitor (CDP). Nfil3 expression is required for the transition from a Zeb2hiand Id2lo stage into a Zeb2lo and Id2hi stage, which has exclusively cDC1 potential. This circuit blocks E protein activity to exclude pDC potential within the progenitor. However how E proteins regulate pDC development is still unclear. Here, we identified an enhancer element 165kb downstream of Zeb2 transcriptional starting site that can be bound by E proteins. Deletion of this enhancer completely abrogates pDC development. Interestingly, the activity of this enhancer is also required for monocytes and B cell development, while tissue resident macrophages used a different enhancer. A circuit involving Zeb2, Id2 and E proteins, where E proteins binding at this cis-element supports Zeb2 expression, is essential for lineage specification at multiple bifurcation point in the hematopoietic tree.
A divergent transcriptional landscape underpins the development and functional branching of MAIT cells
Authors: Koay Hui Fern , Su Shian , Amann-Zalcenstein Daniela , Daley Stephen R , Comerford Iain , Whyte Carly E , Konstantinov Igor E , d’Udekem Yves , Baldwin Tracey , Hickey Peter F , Berzins Stuart P , Mak Jeffrey Y.W. , Kallies Axel , Chen Zhenjun , Nussing Simone , Kedzierska Katherine , Mackay Laura K , McColl Shaun R , Deenick Elissa K , Fairlie David P , McCluskey James , Goodnow Christopher C , Ritchie Matthew E , Belz Gabrielle T , Naik Shalin H , Pellicci Daniel G , Godfrey Dale I 
Published: January 2023

Abstract
MR1-restricted mucosal-associated invariant T (MAIT) cells play a unique role in the immune system. These cells develop intrathymically through a three-stage process, but the events that regulate this are largely unknown. Here, using bulk and single-cell RNA sequencing–based transcriptomic analysis in mice and humans, we studied the changing transcriptional landscape that accompanies transition through each stage. Many transcripts were sharply modulated during MAIT cell development, including SLAM (signaling lymphocytic activation molecule) family members, chemokine receptors, and transcription factors. We also demonstrate that stage 3 “mature” MAIT cells comprise distinct subpopulations including newly arrived transitional stage 3 cells, interferon-γ–producing MAIT1 cells and interleukin-17–producing MAIT17 cells. Moreover, the validity and importance of several transcripts detected in this study are directly demonstrated using specific mutant mice. For example, MAIT cell intrathymic maturation was found to be halted in SLAM-associated protein (SAP)–deficient and CXCR6-deficient mouse models, providing clear evidence for their role in modulating MAIT cell development. These data underpin a model that maps the changing transcriptional landscape and identifies key factors that regulate the process of MAIT cell differentiation, with many parallels between mice and humans.
Activation of AIM2 inflammasome in salivary gland epithelial cells of patients with primary Sjögren’s syndrome
Authors: Hong seungmin , Lee Jaeseon , Jang Se Gwang , Lee Jennifer , Cho Mi-La , Kwok Seung-Ki , Park Sung-Hwan 
Published: January 2023

Abstract
Sjögren’s syndrome (SS) is a chronic and systemic autoimmune disease characterized by lymphocytic infiltration in the exocrine glands. Although an association between inflammasome activation in peripheral blood cells and disease activity of SS has been reported, little is known about inflammasome activation in exocrine glands and its underlying mechanisms. We investigated the levels of AIM2, ASC, caspase-1, IL-1β, and IL-18 in salivary gland tissues and saliva from SS patients. We found that AIM2, ASC, Caspase-1, and IL-18 were increased in the saliva from primary SS patients compared with healthy and sicca controls. Numbers of AIM2-ASC speck also were elevated in the salivary gland epithelial cells of SS patients. The expression of caspase-1 correlated with type I interferon (IFN) signature gene expressions in the minor salivary glands (MSG) of SS patients, and was increased by type I IFN stimulation in salivary gland epithelial cells (SGECs). The AIM2 inflammasome was activated in SGECs after stimulation by the AIM2 ligand, poly(dA:dT). In addition, type I IFN accelerated AIM2 inflammasome activation as determined by the increased expression of caspase-1 in SGECs. In conclusion, the AIM2 inflammasome activation is increased in the SGECs of SS patients, and therefore should be considered an important pathogenic mediator of SS disease.
New classification of CD4+CD8a+ double positive cells in Crohn’s disease patients
Authors: Tanemoto Shun , Sujino Tomohisa , Hagihara Yuya , Takada Yoshiaki , Yoshimatsu Yusuke , Umeda Satoko , Nomura Ena , Ono Keiko , Mikami Yohei , Naganuma Makoto , Kanai Takanori 
Published: January 2023

Abstract
The gastrointestinal tract absorbs nutrition while providing a defense response to foreign antigens including enteric bacteria or food. Once the antigen enter the intestine, inflammatory mononuclear cells are accumulated quickly, however some tissue resident T cells, such as regulatory T cells (Tregs) block the excessive tissue damage. We previously demonstrated that mouse CD4+CD8a+ double expressing (DP)cells and Tregs complementary play an immuno-suppressive role in IE and LP, and a part of IE DP cells is originally developed from Tregs in LP. However, DP cells are not well characterized in human especially in inflammatory bowel disease (IBD) patients.
We obtained human small intestinal tissue from 9 colon cancer patients as normal tissue, and 5 Crohn’s disease(CD) patients, dividing into inflamed and non-inflamed lesion, and analysed by Flow cytometry. We confirmed DP cells are located in both IE and LP in human. First, we demonstrated the ratio of DP cells in normal tissue was as same as in non-inflamed lesion of CD patients. As we expected, the ratio of DP cells in inflamed tissue of CD patients was significantly decreased compared to normal tissue and non-inflamed lesion of CD. Then, we analysed the surface marker of Human DP cells. Surface staining of CD27 and CD45RA showed distinct difference between DP cells and the other CD4 T cells. DP cells expressed more memory T cell marker (CD27−CD45RA−) and less effector T cell marker (CD27+CD45RA−) than the other CD4 subset. Futher more, the more fraction of IE DP cells expressed CD103 than LP DP cells. Conclusively, these data indicated that human DP cell reside both in IE and LP and that DP cells are not a single characterized population, and the DP cells were decreased in inflamed lesion in CD.
Identification of molecules associated with disease progression of autoimmune myocarditis to dilated cardiomyopathy
Authors: Herrera Natalia E , Nalawade Saisha Abhay , Chase Carol Ann , Forsthuber Thomas 
Published: January 2023

Abstract
Myocarditis is an inflammatory disease characterized by immune infiltrates into the myocardium. In many patients, the disease progresses to a chronic phase dilated cardiomyopathy (DCM) in which there is extensive fibrosis leading to ventricular dilation and impaired cardiac function. Less than 50% of these patients survive more than 5 years. Immunosuppressive treatments such as corticosteroids (CSs) are effective in managing the acute phase of the disease, but do not prevent progression to the DCM. Using the experimental autoimmune myocarditis (EAM) animal model, we previously demonstrated that the inflammatory cytokine macrophage migration inhibitory factor (MIF) plays a role in resistance to CS treatment. MIF knockout mice were treated with the corticosteroid Dexamethasone (Dex) and proved to be resistant to both EAM and DCM. We also observed that treatment with MIF inhibitors and Dex decreased the expression of chemokines and adhesion molecules and implicated these molecules in the progression of disease. Within these, osteopontin (OPN), a powerful chemoattractant, was downregulated. We hypothesized that OPN plays a role in the disease process and investigated its role in EAM and DCM using OPN knockout mice. Our results show that OPN knockout mice developed EAM, but that progression to DCM was impaired. Furthermore, we observed a decrease in CD45+CD133+ fibroblast progenitor cells infiltrating the myocardium in the absence of OPN, which could contribute to attenuated disease in KO mice. This study may provide insights into the mechanisms driving progression of EAM to DCM and point to potential therapeutic targets.
Upregulated autophagy in antigen-presenting splenic cells hints at promotion of immune-mediated thrombotic thrombocytopenic purpura (iTTP)
Authors: Schaller Monica , Humbert Magali , Heeb Silvan , Tschan Mario , Hovinga Johanna Kremer 
Published: January 2023

Abstract
Immune-mediated TTP is a life-threatening autoimmune disorder characterized by abundant occlusion of the microcirculation. iTTP is caused by a severe autoantibody-mediated deficiency of the von Willebrand factor cleaving protease, ADAMTS-13. Emerging evidence points to autophagy as key player driving dysregulated autoimmune responses in various diseases. Considering the crucial role of antigen-presenting cells (APCs) in the presentation of autoantigens, we assessed the expression profile of autophagy (ATG)-related genes in iTTP.
We isolated splenic dendritic cells (DC) from one patient carrying the HLA DRB1*11 risk allele and leucocytes from peripheral blood of two healthy donors. Then pan-DCs including plasmacytoid and myeloid DCs were isolated by negative selection followed by FACS sorting. In parallel in vitro differentiated monocyte-derived dendritic cells (Mo-DCs) were cultured from freshly isolated MOs. The expression profile of six essential ATG genes (Beclin-1, Atg5, Atg7, Atg16L1, WIPI1 and LC3B) implicated in autoimmune diseases and the autophagic flux (LC3B expression) were determined by qPCR and detection of LC3B dot formation by immunofluorescence, respectively.
Upregulation of 5 ATG genes (Beclin-1, Atg5, Atg7, WIPI1 and LC3B) was observed in patient’s Mo-DCs compared to controls. Basal autophagy of individual pan-DCs versus Mo-DCs revealed a higher autophagic flux than pan-DCs with the highest autophagic flux displayed in Mo-DCs in the iTTP patient.
The observed activation of autophagy hints at a contribution to the loss of tolerance in iTTP. A more extensive study to investigate if modulators of the respective aberrantly expressed ATG genes might help improve therapy management is ongoing.
Monovalent IgG4 autoantibodies require self-antigen driven affinity maturation to acquire pathogenic capacity
Authors: Fichtner Miriam F L , Vieni Casey , Redler Rachel L. , Jiang Ruoyi , Suarez Pablo , Nowak Richard , Burden Steven J , Bhabha Gira , Ekiert Damian C , O’Connor Kevin C 
Published: January 2023

Abstract

INTRODUCTION
The mechanisms underlying B cell-mediated autoimmune disease and the origin of autoreactive B cells are not well understood. Human monoclonal autoantibodies (mAbs) are valuable tools for investigating both. In this study, we used mAbs derived from patients with the autoimmune disorder, myasthenia gravis (MG). A subset of patients with MG have pathogenic autoantibodies that recognize muscle-specific tyrosine kinase (MuSK). The autoantibodies of MuSK MG are predominantly of the IgG4 subclass and functionally monovalent as a result of Fab-arm exchange.

OBJECTIVE
To gain insight into the origin and development of these unique autoantibodies.

METHODS AND RESULTS
We examined MG patient-derived mAbs, their corresponding germline-encoded unmutated common ancestors (UCA) and monovalent antigen-binding fragments (Fabs) to investigate how antigen-driven affinity maturation contributes to both binding and immunopathology. Mature mAbs, their UCA counterparts and mature monovalent Fabs bound to the MuSK autoantigen and retained their pathogenic capacity. However, monovalent UCA Fabs, which still bound the autoantigen, lost their pathogenic capacity. The mature Fabs were characterized by very high affinity (sub-nanomolar) driven by a rapid on-rate and slow off-rate. However, the UCA affinity was approximately 100-fold less than the mature Fabs, which was driven by a rapid off-rate.

SUMMARY/CONCLUSION
These findings indicate that the autoantigen initiates the autoimmune response in MuSK MG and drives autoimmunity through the accumulation of somatic hypermutations such that monovalent IgG4 Fab-arm exchanged MG autoantibodies reach a high affinity threshold required for pathogenic capacity.
Effects of Morphine on Behavioral and CNS Inflammatory Responses in GP120 Mice Under Immunocompetent vs. Immunodeficient Conditions
Authors: Canonico Dalton , Casale Sadie , Look Tristan , Cao Ling 
Published: January 2023

Abstract
HIV-associated neurocognitive disorder is a common complication of HIV infection, whose development is known to be facilitated by inflammation and exacerbated by morphine. In this study, we infected HIV gp120tg mice with LP-BM5 (a murine retrovirus that can cause systemic immunodeficiency in susceptible mouse strains) to examine morphine’s effect on gp120 in both immunocompetent and immunodeficient conditions. Specifically, we examined the expression of a series of inflammation-related molecules in the brain via qRT-PCR as well as the animals’ learning/memory function using spontaneous alternation T-maze. Morphine treatment (2×25mg pellets) did not significantly affect the development of immunodeficiency induced by LP-BM5 and all brain regions examined (hippocampus, striatum, and frontal lobe) had detectable LP-BM5 viral gag genes. Morphine notably reduced the performance of gp120tg+ mice in the T-maze assay when 2-minute retention was used, regardless of LP-BM5 treatment. Without LP-BM5 treatment, morphine increased hippocampal expression of CCL2, CCL5, CXCL10, IL-12 p40, TNFα, and IFNγ, while it decreased the expression of IL-6, arginase-1, IFNα and IFNβ. For mice who were subjected to LP-BM5 treatment, hippocampal expression of CCL2, CCL5, CXCL10, IL-12p40, and IL-1β were decreased by morphine, while CCL4, TNFα, IFNγ, iNOS, arginase-1 and IFNβ were increased by morphine. Further, expression of the mu opioid receptor was upregulated by morphine without LP-BM5, yet down-regulated by morphine with LP-BM5 treatment. Altogether, the effects of morphine are complex and dependent on the immune status of the host, which will be further elucidated in the future studies. (Supported by NIH/NIDA R21DA044886 (PI, Cao))
Intracellular expression of IFN-λ4 leads to ER stress, enhanced IRF1 signaling and decreased proliferation in hepatic cells that might protect HCV+ patients from liver cirrhosis
Authors: Onabajo Olusegun O , Wang Fang , Lee Mei-Hsuan , Florez-Vargas Oscar , Obajemu Adeola , Castro Mauro A , Vargas Joselin , Liao Shu-Fen , Huang Yu-Han , Shen Chen-Yang , Banday Rouf , O’Brien Thomas , Robertson A. Gordon , Prokunina-Olsson Ludmila 
Published: January 2023

Abstract
Several genetic variants in the human IFNL3/IFNL4 locus have been associated with reduced hepatic fibrosis despite poor clearance of HCV infection. We expanded this analysis to cirrhosis, a more advanced stage of fibrosis. In 2931 individuals with chronic HCV, the IFNL4 genotype that generates IFN-λ4 was associated with protection from cirrhosis (OR=0.65, p=0.012, adjusted for age and sex). The IFNL4 genotype affects the production of IFN-λ4 and, additionally, may affect expression levels of IFN-λ3, making it difficult to delineate the individual contribution of these IFNs. To address this, we established HepG2-based cell models engineered to inducibly express either IFN-λ3 or IFN-λ4. Using RNA-seq based expression profile generated in these hepatic cell lines, we explored the global transcriptome of liver tumors (n=373) from The Cancer Genome Atlas to identify transcription factor networks affected by these IFNs in the liver. Several networks, including IRF1, were upregulated by IFN-λ4 more strongly than by IFN-λ3. Intracellular expression of IFN-λ4 but not of IFN-λ3 also led to potent IRF1-dependent antiproliferative effects. Live cell imaging revealed that IFN-λ4 was poorly secreted, mainly accumulated in lysosomes, and caused apoptosis, suggesting increased ER-stress via the misfolded protein response. Knockdown of DNA damage-inducible transcript 3 (DDIT3), an ER-stress response effector, significantly attenuated the antiproliferative effects of IFN-λ4. This novel interplay of enhanced IRF1 signaling coupled with intracellular accumulation and induction of ER stress by IFN-λ4 may have complex consequences on liver homeostasis during chronic HCV infection but also mediate anti-cirrhotic phenotypes.