B cells are capable of independently eliciting rapid reactivation of encephalitogenic CD4 T cells in a murine model of multiple sclerosis

<div><p>Recent success with B cell depletion therapies has revitalized efforts to understand the pathogenic role of B cells in Multiple Sclerosis (MS). Using the adoptive transfer system of experimental autoimmune encephalomyelitis (EAE), a murine model of MS, we have previously shown that mice in which B cells are the only MHCII-expressing antigen presenting cell (APC) are susceptible to EAE. However, a reproducible delay in the day of onset of disease driven by exclusive B cell antigen presentation suggests that B cells require optimal conditions to function as APCs in EAE. In this study, we utilize an <i>in vivo</i> genetic system to conditionally and temporally regulate expression of MHCII to test the hypothesis that B cell APCs mediate attenuated and delayed neuroinflammatory T cell responses during EAE. Remarkably, induction of MHCII on B cells following the transfer of encephalitogenic CD4 T cells induced a rapid and robust form of EAE, while no change in the time to disease onset occurred for recipient mice in which MHCII is induced on a normal complement of APC subsets. Changes in CD4 T cell activation over time did not account for more rapid onset of EAE symptoms in this new B cell-mediated EAE model. Our system represents a novel model to study how the timing of pathogenic cognate interactions between lymphocytes facilitates the development of autoimmune attacks within the CNS.</p></div

Music-based language programme teacher training on preschool teachers’ music teaching self-efficacy and delivery performance : A case study.

Music substantially impacts early childhood development, especially language development, yet early childhood policies rarely emphasise the role of music in early learning and engagement. Research shows that effective delivery of early childhood music programmes depends on teachers' music knowledge, skills, and music teaching self-efficacy, which may be enhanced through teacher training. However, little is known about the training details and preparation of non-music specialist preschool teachers who use music to teach. The present study sought to determine whether music-based language programme (MBLP©) teacher training could improve the music teaching self-efficacy of non-music specialist preschool teachers and their MBLP© delivery in an inclusive preschool classroom. The 14 training sessions consist of a briefing, a workshop, a practicum, interactive reflections, and observations. Three non-music-specialist preschool teachers were trained to deliver MBLP© lessons to nine preschoolers aged 33 to 47 months. The within-subjects design investigated the teachers' self-efficacy in music teaching, while the single-subject design tracked their MBLP© lesson delivery performance. Data was collected using a self-reported music background survey, pre-, mid-, and post-test music teaching self-efficacy, teachers' feedback, and MBLP© lesson observations. The results showed that the training increased music teaching selfefficacy by 10 to 46% and revealed a positive relationship between practicum with interactive reflections and lesson delivery performance. The findings suggest that providing MBLP© training to non-specialist preschool teachers could help address language developmental issues in inclusive preschool settings

Music-based Language Programme for Preschool Teachers' Training and Lesson delivery: A Pilot Trial

Language delay in preschoolers is linked to learning difficulties and persistent communication issues. Studies show that early music-making positively impacts young children's language development. Music-based Language Programme aims to enhance preschoolers’ language skills by training preschool teachers to deliver quality music lessons with language-focused objectives. This pilot trial aimed to determine the feasibility of teacher training for the programme's delivery. Furthermore, the secondary aim was to evaluate the acceptability of the programme's implementation by stakeholders. Methods: The 6-week one-arm pilot trial took place in Sarawak, Malaysia (October to November 2021). It included preschool visits, parent briefings, teacher training, and music lesson delivery. The participants included teachers (n = 4), preschoolers (n = 11), parents (n = 11), and the principal (n = 1) in a preschool that practiced inclusion. A non-probability-purposive sampling recruitment strategy was employed amid the COVID-19 pandemic. The study used a case study mixed methods approach to collect data through reports, observations, group discussions, feedback, questionnaires, and documentation. Results: There was a high rate of participant retention (100%) and completion of data collection tasks (91–100%). Additionally, there was a favourable shift (7–27%) in three teachers' self-efficacy ratings before and after the teacher training and a reasonably high implementation fidelity (87%). There were no adverse events related to the study participants. Conclusion: The study demonstrated promising results across multiple participant levels, as it was perceived to be feasible, acceptable, and appropriate by teachers, preschoolers, the principal, and parents. The findings provided direct implications for the progression of the pilot trial to the full-scale main study

Regional CNS responses to IFN-γ determine lesion localization patterns during EAE pathogenesis

The localization of inflammatory foci within the cerebellum is correlated to severe clinical outcomes in multiple sclerosis (MS). Previous studies of experimental autoimmune encephalomyelitis (EAE), a model of MS, revealed distinct clinical outcomes correlated with the capacity of the animal to produce IFN-γ. Outcomes were linked to localization of inflammatory cells in either the spinal cord (wild type [WT]) or the cerebellum and brain stem (IFN-γ deficient). We demonstrate, using an adoptive transfer system, that the ability of the central nervous system (CNS) to sense pathogenic T cell–produced IFN-γ during EAE initiation determines the sites of CNS pathogenesis. Transfer of WT Th1 cells into IFN-γ receptor–deficient mice results in pathogenic invasion of the brain stem and cerebellum with attendant clinical symptoms, which are identical to the disease observed after transfer of IFN-γ–deficient T cells to WT hosts. Inflammation of the spinal cord associated with classical EAE is abrogated in both IFN-γ–deficient systems. Cotransfer of CNS antigen-specific WT Th1 cells with IFN-γ–deficient T cells is sufficient to restore spinal cord invasion and block cerebellar and brain stem invasion. These data demonstrate that interaction between IFN-γ and host CNS cells during the initiation of EAE can selectively promote or suppress neuroinflammation and pathogenesis

Synthetic Text Generation with Differential Privacy: A Simple and Practical Recipe

Privacy concerns have attracted increasing attention in data-driven products due to the tendency of machine learning models to memorize sensitive training data. Generating synthetic versions of such data with a formal privacy guarantee, such as differential privacy (DP), provides a promising path to mitigating these privacy concerns, but previous approaches in this direction have typically failed to produce synthetic data of high quality. In this work, we show that a simple and practical recipe in the text domain is effective: simply fine-tuning a pretrained generative language model with DP enables the model to generate useful synthetic text with strong privacy protection. Through extensive empirical analyses on both benchmark and private customer data, we demonstrate that our method produces synthetic text that is competitive in terms of utility with its non-private counterpart, meanwhile providing strong protection against potential privacy leakages.Comment: ACL 2023 Main Conference (Honorable Mention

Engineering novel complement activity into a pulmonary surfactant protein

Complement neutralizes invading pathogens, stimulates inflammatory and adaptive immune responses, and targets non- or altered-self structures for clearance. In the classical and lectin activation pathways, it is initiated when complexes composed of separate recognition and activation subcomponents bind to a pathogen surface. Despite its apparent complexity, recognition-mediated activation has evolved independently in three separate protein families, C1q, mannose-binding lectins (MBLs), and serum ficolins. Although unrelated, all have bouquet-like architectures and associate with complement-specific serine proteases: MBLs and ficolins with MBL-associated serine protease-2 (MASP-2) and C1q with C1r and C1s. To examine the structural requirements for complement activation, we have created a number of novel recombinant rat MBLs in which the position and orientation of the MASP-binding sites have been changed. We have also engineered MASP binding into a pulmonary surfactant protein (SP-A), which has the same domain structure and architecture as MBL but lacks any intrinsic complement activity. The data reveal that complement activity is remarkably tolerant to changes in the size and orientation of the collagenous stalks of MBL, implying considerable rotational and conformational flexibility in unbound MBL. Furthermore, novel complement activity is introduced concurrently with MASP binding in SP-A but is uncontrolled and occurs even in the absence of a carbohydrate target. Thus, the active rather than the zymogen state is default in lectin·MASP complexes and must be inhibited through additional regions in circulating MBLs until triggered by pathogen recognition

Batf3-Dependent CD11blow/− Peripheral Dendritic Cells Are GM-CSF-Independent and Are Not Required for Th Cell Priming after Subcutaneous Immunization

Dendritic cells (DCs) subsets differ in precursor cell of origin, functional properties, requirements for growth factors, and dependence on transcription factors. Lymphoid-tissue resident CD8α+ conventional DCs (cDCs) and CD11blow/−CD103+ non-lymphoid DCs are developmentally related, each being dependent on FMS-like tyrosine kinase 3 ligand (Flt3L), and requiring the transcription factors Batf3, Irf8, and Id2 for development. It was recently suggested that granulocyte/macrophage colony stimulating factor (GM-CSF) was required for the development of dermal CD11blow/−Langerin+CD103+ DCs, and that this dermal DC subset was required for priming autoreactive T cells in experimental autoimmune encephalitis (EAE). Here, we compared development of peripheral tissue DCs and susceptibility to EAE in GM-CSF receptor deficient (Csf2rb−/−) and Batf3−/− mice. We find that Batf3-dependent dermal CD11blow/−Langerin+ DCs do develop in Csf2rb−/− mice, but that they express reduced, but not absent, levels of CD103. Further, Batf3−/− mice lacking all peripheral CD11blow/− DCs show robust Th cell priming after subcutaneous immunization and are susceptible to EAE. Our results suggest that defective T effector priming and resistance to EAE exhibited by Csf2rb−/− mice does not result from the absence of dermal CD11blow/−Langerin+CD103+ DCs

Energy flux couples sulfur isotope fractionation to proteomic and metabolite profiles in Desulfovibrio vulgaris

Funding Information: We thank S. Moore and D. Fike for bulk sulfur isotope analyses (WashU); M. Seuss for assistance with lipid\u2010H isotope analyses (Bradley lab, WashU); X. Feng (Dartmouth) and M. Osburn (Northwestern) for water H\u2010isotope analyses; and A. Sessions and J. Adkins (CalTech) for access to HPLC\u2010ICP\u2010MS. Metabolite analyses were performed by the Proteomics & Mass Spectrometry Facility at the Danforth Plant Science Center (St. Louis, MO, USA). Funding was provided: by NASA Exobiology Award 13\u2010EXO13\u20100082 (ASB, WDL, JW), NSF\u2010EAR Award 1928309 (WDL), Washington University in St. Louis Department of Earth & Planetary Sciences Fossett Fellowship (WDL), the Walter and Constance Burke Fund at Dartmouth College (WDL), and the Fulbright\u2014Bunge & Born\u2014Williams Foundation Scholarship Program (FJB), Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia (Portugal) through R&D unit MOSTMICRO\u2010ITQB (UIDB/04612/2020 and UIDP/04612/2020) and LS4FUTURE Associated Laboratory (LA/P/0087/2020) (IACP), NSF GRFP [2017250547] (SP). Funding Information: We thank S. Moore and D. Fike for bulk sulfur isotope analyses (WashU); M. Seuss for assistance with lipid-H isotope analyses (Bradley lab, WashU); X. Feng (Dartmouth) and M. Osburn (Northwestern) for water H-isotope analyses; and A. Sessions and J. Adkins (CalTech) for access to HPLC-ICP-MS. Metabolite analyses were performed by the Proteomics & Mass Spectrometry Facility at the Danforth Plant Science Center (St. Louis, MO, USA). Funding was provided: by NASA Exobiology Award 13-EXO13-0082 (ASB, WDL, JW), NSF-EAR Award 1928309 (WDL), Washington University in St. Louis Department of Earth & Planetary Sciences Fossett Fellowship (WDL), the Walter and Constance Burke Fund at Dartmouth College (WDL), and the Fulbright\u2014Bunge & Born\u2014Williams Foundation Scholarship Program (FJB), Funda\u00E7\u00E3o para a Ci\u00EAncia e Tecnologia (Portugal) through R&D unit MOSTMICRO-ITQB (UIDB/04612/2020 and UIDP/04612/2020) and LS4FUTURE Associated Laboratory (LA/P/0087/2020) (IACP), NSF GRFP [2017250547] (SP). Publisher Copyright: © 2024 The Authors. Geobiology published by John Wiley & Sons Ltd.Microbial sulfate reduction is central to the global carbon cycle and the redox evolution of Earth's surface. Tracking the activity of sulfate reducing microorganisms over space and time relies on a nuanced understanding of stable sulfur isotope fractionation in the context of the biochemical machinery of the metabolism. Here, we link the magnitude of stable sulfur isotopic fractionation to proteomic and metabolite profiles under different cellular energetic regimes. When energy availability is limited, cell-specific sulfate respiration rates and net sulfur isotope fractionation inversely covary. Beyond net S isotope fractionation values, we also quantified shifts in protein expression, abundances and isotopic composition of intracellular S metabolites, and lipid structures and lipid/water H isotope fractionation values. These coupled approaches reveal which protein abundances shift directly as a function of energy flux, those that vary minimally, and those that may vary independent of energy flux and likely do not contribute to shifts in S-isotope fractionation. By coupling the bulk S-isotope observations with quantitative proteomics, we provide novel constraints for metabolic isotope models. Together, these results lay the foundation for more predictive metabolic fractionation models, alongside interpretations of environmental sulfur and sulfate reducer lipid-H isotope data.publishersversionpublishe

Energy flux couples sulfur isotope fractionation to proteomicand metabolite profiles in Desulfovibrio vulgaris

Fil: Leavitt, William D. Dartmouth College. Department of Earth Sciences, Hanover, New Hampshire; United States of America.Fil: Leavitt, William D. Washington University in St. Louis. Department of Earth and Planetary Sciences, Missouri; United States of America.Fil: Waldbauer, Jacob. University of Chicago. Department of the Geophysical Sciences, Illinois; United States of America.Fil: Venceslau, Sofia S. Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica António Xavier, Oeiras; Portugal.Fil: Sim, Min Sub. Seoul National University. School of Earth and Environmental Sciences; South Korea.Fil: Zhang, Lichun. University of Chicago. Department of the Geophysical Sciences, Illinois; United States of America.Fil: Flavia Jaquelina Boidi. Washington University in St. Louis. Department of Earth and Planetary Sciences; Missouri, United States of America.Fil: Flavia Jaquelina Boidi. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Investigaciones en Ciencias de la Tierra; Argentina.Fil: Flavia Jaquelina Boidi. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Plummer, Sydney. University of California, San Diego, La Jolla. Scripps Institution of Oceanography, California; United States of America.Fil: Díaz, Julia M. University of California, San Diego, La Jolla. Scripps Institution of Oceanography, California; United States of America.Fil: Pereira, Inês A. C. Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica António Xavier, Oeiras; Portugal.Fil: Bradley, Alexander S. Washington University in St. Louis. Department of Earth and Planetary Sciences; Missouri, United States of America.Fil: Bradley, Alexander S. Washington University in St. Louis. Division of Biology and Biomedical Sciences, Missouri; United States of America.Abstract: Microbial sulfate reduction is central to the global carbon cycle and the redox evolu-tion of Earth's surface. Tracking the activity of sulfate reducing microorganisms overspace and time relies on a nuanced understanding of stable sulfur isotope fractiona-tion in the context of the biochemical machinery of the metabolism. Here, we link themagnitude of stable sulfur isotopic fractionation to proteomic and metabolite profilesunder different cellular energetic regimes. When energy availability is limited, cell-specific sulfate respiration rates and net sulfur isotope fractionation inversely covary. Beyond net S isotope fractionation values, we also quantified shifts in protein expres-sion, abundances and isotopic composition of intracellular S metabolites, and lipidstructures and lipid/water H isotope fractionation values. These coupled approachesreveal which protein abundances shift directly as a function of energy flux, those thatvary minimally, and those that may vary independent of energy flux and likely do notcontribute to shifts in S-isotope fractionation. By coupling the bulk S-isotope obser-vations with quantitative proteomics, we provide novel constraints for metabolic iso-tope models. Together, these results lay the foundation for more predictive metabolicfractionation models, alongside interpretations of environmental sulfur and sulfatereducer lipid-H isotope data.info:eu-repo/semantics/publishedVersionFil: Leavitt, William D. Dartmouth College. Department of Earth Sciences, Hanover, New Hampshire; United States of America.Fil: Leavitt, William D. Washington University in St. Louis. Department of Earth and Planetary Sciences, Missouri; United States of America.Fil: Waldbauer, Jacob. University of Chicago. Department of the Geophysical Sciences, Illinois; United States of America.Fil: Venceslau, Sofia S. Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica António Xavier, Oeiras; Portugal.Fil: Sim, Min Sub. Seoul National University. School of Earth and Environmental Sciences; South Korea.Fil: Zhang, Lichun. University of Chicago. Department of the Geophysical Sciences, Illinois; United States of America.Fil: Flavia Jaquelina Boidi. Washington University in St. Louis. Department of Earth and Planetary Sciences; Missouri, United States of America.Fil: Flavia Jaquelina Boidi. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Investigaciones en Ciencias de la Tierra; Argentina.Fil: Flavia Jaquelina Boidi. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Plummer, Sydney. University of California, San Diego, La Jolla. Scripps Institution of Oceanography, California; United States of America.Fil: Díaz, Julia M. University of California, San Diego, La Jolla. Scripps Institution of Oceanography, California; United States of America.Fil: Pereira, Inês A. C. Universidade Nova de Lisboa. Instituto de Tecnologia Química e Biológica António Xavier, Oeiras; Portugal.Fil: Bradley, Alexander S. Washington University in St. Louis. Department of Earth and Planetary Sciences; Missouri, United States of America.Fil: Bradley, Alexander S. Washington University in St. Louis. Division of Biology and Biomedical Sciences, Missouri; United States of America

Bhlhe40 controls cytokine production by T cells and is essential for pathogenicity in autoimmune neuroinflammation

T(H)1 and T(H)17 cells mediate neuroinflammation in experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis. Pathogenic T(H) cells in EAE must produce the pro-inflammatory cytokine granulocyte-macrophage colony stimulating factor (GM-CSF). T(H) cell pathogenicity in EAE is also regulated by cell-intrinsic production of the immunosuppressive cytokine interleukin 10 (IL-10). Here, we demonstrate that mice deficient for the basic helix-loop-helix (bHLH) transcription factor Bhlhe40 (Bhlhe40(−/−)) are resistant to the induction of EAE. Bhlhe40 is required in vivo in a T cell-intrinsic manner, where it positively regulates the production of GM-CSF and negatively regulates the production of IL-10. In vitro, GM-CSF secretion is selectively abrogated in polarized Bhlhe40(−/−) T(H)1 and T(H)17 cells, and these cells show increased production of IL-10. Blockade of IL-10 receptor in Bhlhe40(−/−) mice renders them susceptible to EAE. These findings identify Bhlhe40 as a critical regulator of autoreactive T cell pathogenicity