bluuebunny/crossref_metadata_2025 huggingface dataset:
The key to bringing DNA collections to the next level
Authors: Veltjen Emily , Asselman Pieter , Baert Wim , Baeyen Steve , Beirinckx Lise , Breyne Liselot , Brosens Dimitri , Claerhout Tim , Cogneau Sari , Cox Karen , Cuypers Laura , Delgat Lynn , Desmeth Philippe , de Raad Jordi , Esselens Lore , Eves Down Maria-Rose , Helsen Philippe , Leliaert Frederik , Meganck Kenny , Pereboom Zjef , Smitz Nathalie , Sonet Gontran , Trekels Maarten , Vanden Broeck An , Van Driessche Charlotte , De Wever Aaike
Published: November 2024

DNA collections are a valuable type of Natural Science collection, enabling the validation of past research, serving as a source for new genomic studies and supporting ex situ conservation. The DiSSCo Flanders DNA collection working group, aiming to advance and "unlock" their DNA collections, identified the need for: 1) actively sharing best practices regarding the management of DNA collections; and 2) providing guidance on how to bring theory into practice. By combining best practice examples from within the working group with available literature and brainstorming ideas, the working group co-created two outputs, referred to as: the "Challenges" and the "Key". The Challenges are a list of obstacles to DNA collection management, which shape the structure of the linked Key and can also be used to spark discussion amongst stakeholders. The Key is a tool that guides users through the maturation process of their DNA collection in a standardised way. It stimulates holistic growth, breaks down the needed work into manageable steps and helps to decide priorities during the process. Furthermore, the Key facilitates communication with both internal stakeholders and external DNA collection managers. The Key distinguishes itself from other self-assessment tools in several ways: it includes (re)investigation of the collection’s purpose and context; it is specialised for DNA collections; it delivers concrete goals linked to relevant information and shared experience; and it is inclusive, targeting all Natural Science DNA collections, regardless of their context or size.
Three new species of the genus Bambusiphaga Huang & Ding, 1979 (Hemiptera, Fulgoroidea, Delphacidae, Tropidocephalini) from China, with an updated checklist and key to species
Authors: Lv Sha-Sha , Li Hong-Xing , Yang Lin , Zhang Yu-Bo , Chen Xiang-Sheng
Published: November 2024

In this study, three new bamboo-feeding species of the genus Bambusiphaga Huang & Ding, 1979 (Hemiptera, Fulgoroidea, Delphacidae, Tropidocephalini), B. caudospina Lv, Li & Chen, sp. nov., B. laterospina Lv, Li & Chen, sp. nov. and B. striola Lv, Li & Chen, sp. nov. from Southwest China, are described and illustrated, bringing the total number of species in the genus to 34. An updated identification key and checklist to all known species of Bambusiphaga are provided.
All Around the Wrekin: The Midlands as (M)otherland in Liz Berry’s Poetry
Authors: Braun Alice , Hélie Claire , Trainor Samuel
Published: November 2024

This article explores the themes of place and transition in the poetry of Liz Berry, especially in Black Country (2014) and The Republic of Motherhood (2018). Berry is an avid reader of the major figures of Midlands poetry, like Roy Fisher, and works closely with other local poets and artists, like R. M. Francis and Tom Hicks. Her poems inhabit and rechart the landscape of the West Midlands, its industrial past, its ecological and artistic transformations. However, her work extends beyond the geographical to explore another land, that of motherhood. Ultimately, her voice breathes new life into the prosodic landscapes of the Black Country accent during the performances of her poems.
Due North? British Nature Writing Prizes: Reworking Cardinal Points in Contemporary British Nature Writing
Authors: Beaufils Cécile
Published: November 2024

Recent publications belonging to British nature writing have been marked by an evolution in the representation of space and geography. Some authors have focused on reworking boundaries and direction, for instance in 2012 Poet Laureate Simon Armitage insisted in Walking Home that he should walk “in the wrong direction”, or Robert Macfarlane, who in 2012 partly structured The Old Ways according to the cardinal points which directed his walking explorations. Other authors choose to rework tropes belonging to the use of fixed points of reference in space by exploring directions as set markers, like Melissa Harrison with All Among the Barley (2018), which is set in East Anglia, or Sarah Hall in The Wolf Border (2015). As stressed by Abberley et al. in Land Lines it cannot be assumed that nature and nature writing work as markers for conservative ideals either, and it can be argued that such recent works have done much to reinvent nature writing as a genre which can exist with a strong sense of directional markers and cardinal points and a distinct aesthetics which points at connectedness. To examine the representation of such directions, we will pay particular attention to recent publications, and more specifically to two texts which were each awarded a recent literary prize (given the prolific quality of the field, readers have required an increase in orientation themselves): Uprooting (Marchelle Farrell 2023), which won the Nan Shepherd Prize in 2021, and English Pastoral (James Rebanks 2021), which won the Wainwright Prize the same year. We will then endeavour to observe the structuring role of cardinal points in these texts and their place in a quickly-evolving literary field.
L’occupation du haut Moyen Âge (ive-xe s.) des Huguetières à La Chevrolière (Loire-Atlantique) : la composante économique d’un domaine ?
Authors: Peytremann Édith , Forré Philippe , Poilpré Pierre , Wiethold Julian , Zaour Nolwenn
Published: November 2024

L’occupation alto-médiévale du site des Huguetières à La Chevrolière, en Loire-Atlantique, s’inscrit dans la continuité des précédentes, datées des périodes pré- et protohistoriques et de l’Antiquité. Elle se caractérise par la présence de deux enclos palissadés monumentaux et d’un atelier sidérurgique où sont attestées les différentes étapes du système de production, du minerai à la forge. La singularité des infrastructures ainsi que les indices matériels (lampes à huile, meules exogènes, ardillon damasquiné de laiton, etc.) posent la double question de la survivance sur une longue période d’un ensemble foncier, et de l’appartenance sociale des propriétaires. Ce site est en effet localisé à proximité d’une villa gallo-romaine et au sein d’un domaine foncier correspondant au siège de la châtellenie des Huguetières, dont l’existence, attestée depuis le xiiie s., pourrait se révéler bien antérieure à ses premières mentions écrites.
Exposições de cosmorama e o museu: um projecto para reimaginar futuros de uma história esquecida
Authors: Martins Susana S.
Published: November 2024

Este artigo examina a relação entre os museus e a história esquecida do Cosmorama, uma prática imersiva de visualização de imagens que se popularizou nas cidades ocidentais na primeira metade do século XIX, sobretudo sob a forma de exposições. Apresentados principalmente em galerias urbanas e, pontualmente, em alguns museus da época, poucos vestígios materiais desta cultura óptica sobreviveram nas colecções museológicas de hoje, contribuindo tanto para a sua ausência generalizada do campo da história das exposições, como para a sua “perda” museológica. Este texto tem por objectivo redefinir o lugar museológico das exposições de cosmorama a partir da apresentação dos objectivos e desenvolvimentos do projecto de investigação “CURIOSITAS: Espreitar antes da Realidade Virtual. Uma Arqueologia dos Media Imersivos através da Realidade Virtual e dos Cosmoramas Ibéricos” (2022-2025) que estuda estas exposições numa perspectiva de humanidades digitais. Numa primeira fase, são identificadas características para-museológicas dos cosmoramas e discutidas hipóteses que podem explicar a sua atual invisibilidade museológica. Subsequentemente, descreve-se o alcance internacional e os princípios metodológicos que orientam este projecto, propondo-se novas soluções possíveis para reavaliar e reintegrar este património esquecido nas práticas e discursos museológicos de hoje e do futuro.
El medio es el mensaje. Museología y pinkwashing en la era digital
Authors: Barrio Javier Cuevas del
Published: November 2024

El tema de este artículo se centra en la inclusión de la diversidad sexo-afectiva en las exposiciones y colecciones permanentes de museos. Hemos tomado como caso de estudio la exposición sobre el artista italiano Guido Reni (1575-1642) en el Museo Nacional del Prado de Madrid (marzo-julio 2023). Para ello, además de visitar la exposición, hemos analizado los nueve ensayos publicados en el amplio catálogo de la exposición, así como la difusión a través de redes sociales, podcast y otras vías del contenido de esta. En ese sentido, queremos demostrar cómo algunas estrategias de inclusión pueden funcionar también como estrategias de exclusión e invisibilización. Dichas estrategias también pueden quedar reducidas a una política de pinkwashing, es decir, de “lavado rosa” consistente en estrategias políticas y de marketing que consiguen atraer el turismo LGBTIQ+ en el marco de las fiestas del Orgullo, y que museos como el Prado sean vistos como instituciones simpatizantes de los derechos civiles de las personas LGBTIQ+, y, por lo tanto, instituciones progresistas, modernas y tolerantes.
Geopolymers Prepared From Unused Resources and Their Applications
Authors: Nikaido Yuta Watanabe , Onutai Sujitra , Jiemsirilers Sirithan , Kobayashi Takaomo
Published: November 2024

As an alumino-silicate material, geopolymers present several of beneficial properties, including high strength, low permeability, high acid resistance, the capacity to immobilize poisonous compounds, and the ability to resist hazardous contaminants. In this chapter, geopolymer materials using raw materials derived from unutilized waste are discussed, especially the case of using metakaolin-based and fly ash-based raw materials, and the preparation and properties of geopolymers and their applications are explained. The main point is that geopolymers are materials that can contribute to the construction of a sustainable society as a technology that can create high value-added products through the effective use of unused waste materials.
Effect of Heat Treatment on Impact Transition Temperature of High Strength Low Alloy Steel (HSLA) Steel Rods
Authors: Akhtar Shahid
Published: November 2024

This study is carried out to investigate the ductile to brittle fracture behavior of special high strength low alloy steel (HSLA), which is used in aerospace applications. As aerospace materials are subjected to temperature variations during their actual use therefore it is very important to know their fracture behavior with change of temperature. Alloy having 0.3 %C, 0.88 % Mn and 1.0% Si was heat treated to obtain tempered martensite and ferrite and tempered martensite structures. Impact toughness tests were carried out at temperature range from – 60 °C - 200 °C in as received, fully quenched, and tempered and quenched and tempered in two phase (a+ γ) state. Then the effect of different heat treatments on impact transition temperature was studied.
Bifunctional Fe2WO6/SrTiO3 Composites with Eminent Antibacterial and Photocatalytic Studies
Authors: Lam Sze Mun , Sin Jin Chung , Zhao Liang , Zeng Hong Hu
Published: November 2024

Decorated Fe2WO6 onto SrTiO3 nanocubes (Fe2WO6/SrTiO3) composites were fabricated by a two-step hydrothermal technique. Some spectroscopic techniques were adopted to delve the catalyst samples. The composite manifested a prominent antibacterial effect toward Escherichia Coli and Bacillus cereus bacteria. Importantly, the as-fabricated composite was testified to exhibit good photodegradation of chlortetracycline hydrochloride under visible light. The eminent performance of the composite was ascribed to not only the excellent absorption of visible light, but also due to the synergistic effect of S-scheme heterojunction which endowed for the high yield of the active species for the pollutant and microorganism destruction. Our study implied that Fe2WO6/SrTiO3 composites can be a potential bifunctional option for remediating microbial-polluted water.
Considerations on the Behaviour and Resistance of Aluminium Alloys to Cavitation Erosion
Authors: Bordeasu Ilare , Luca Alexandru Nicolae , Ghera Cristan , Salcianu Cornelia Laura , Ostoia Daniel , Micu Lavinia Madalina
Published: November 2024

Aluminum alloys are known for their wide application in the automotive, river and marine boat constructions, but also in hydraulic systems (radiators/oil coolers). Their use is made by manufacturing parts directly from the semi-finished state, with or without certain volumetric heat treatments and surface hardening, depending on the functional role and the physical-mechanical characteristics pursued. Some of these parts work in hydrodynamic conditions, where cavitation manifests itself through erosion, such as: propellers of boats and barges, pump rotors in water cooling systems of automobiles. Visual analyzes performed on sailboat and powerboat propellers, after identical durations and operating conditions, showed cavitation erosion damage, different depending on the type of aluminum alloy. As a result, the paper presents and analyzes the behavior and resistance to erosion by vibratory cavitation of 4 types of aluminum alloys in the state of rolled semi-finished products. To highlight the differences in the destruction of structures under the cyclic stresses of cavitational microjets, macro and microscopic images of the eroded structure are used, as well as the curves with the values of the parameters specific to cavitation, recommended by the ASTM G32 -2016 norms and used in the research laboratory's custom. The analysis of the results shows that the resistance of the structure to cavitation erosion is dependent on the type of alloy, the degree of brittle intermetallic compounds and the mechanical properties specific to toughness.
αPDL2 treats melanoma in an IL17 and IFNγ dependent mechanism mediated by CD8 and γδ T cells
Authors: Tanner Chris , Ontiveros Carlos , Bai Haiyan , Garcia Myrna
Published: November 2024

Immune checkpoint blockade (ICB) strategies have transformed immunotherapy with demonstrated efficacy including in melanoma. Nonetheless, most tumors do not respond to approved ICB agents. PDL2 is a relatively understudied immune checkpoint molecule that binds PD1 with higher affinity than PDL1, and also binds RGMb. We found that αPDL2 effectively treats subcutaneous B16 and Nras-mutated melanomas in aged but not young mice. αPDL2 efficacy in aged B16 requires host IL17 which induces IFNγ and is mediated by both γδ and CD8+ T cells. IL17 increased CXCR3+IFNγ+ immune cells, also supporting improved trafficking as a mechanism. αPDL2 augments B16-infiltrating γδ T cell numbers in aged mice, which boost tumor-infiltrating CD8+ T cell functions (e.g., IFNγ[TC1] ) but not numbers. CD8+ T cells did not appear to improve δγ T cell functions or numbers. αPDL2 also increased tumor infiltrating NK cells, CD4+ T cells, and dendritic cells that can mediate anti-tumor immunity. αPDL2 effectively treats heterotopic subcutaneous MB49 bladder cancer in young and aged mice showing effects are not restricted to only aged. Furthermore, lack of tumor PDL2 expression did not alter αPDL2 efficacy observed. PDL2 is thus a viable ICB target for treatment of cancer in select patient populations, including the aged.
Inducing Neoepitopes in Cancer Cells Unmasks Tumors to the Immune System
Authors: Volpatti Lisa , Borjas Gustavo , Rosenthal Joshua , Hubbell Jeffrey
Published: November 2024

Immunotherapy with immune checkpoint inhibitors (ICIs) is limited to a subset of cancer types. The treatment of immunologically cold tumors that lack T cell infiltration remains a challenge. Since tumor mutational burden correlates with ICI efficacy, here our objective is to artificially induce “mutations” in cancer cells at the level of RNA to unmask cold tumors to the immune system and render them susceptible to ICI. To accomplish this, we engineered human Adenosine Deaminase that Acts on RNA 2 (ADAR2) to create abundant, genome-wide editing events. We hypothesized that these edits would induce neoepitopes and unmask tumors to the immune system. We tested this hypothesis in immunologically cold murine melanoma (B16F10). Mice received one injection of adeno-associated viruses (AAVs) encoding ADAR2 in combination with αPD1. Tumor growth was suppressed upon treatment, resulting in a significant improvement in overall survival compared to αPD1 monotherapy. Flow cytometry analysis of the tumor revealed significantly increased CD8+ T cell infiltration and tumor-specific T cells as well as a reduction of regulatory T cells and myeloid-derived suppressor cells. In an immune excluded model of breast cancer (EMT6), we achieve complete remission of established tumors in a quarter of treated mice. Thus, inducing neoepitopes through RNA editing may be a novel method of enhancing immunotherapy by unmasking a variety of cold tumors to the immune system.
Developing thymocytes cue in unique subsets of neurons to support optimal thymic architecture
Authors: Hoyt-Miggelbrink Alexandra , Pfaller Christian , Yokanovich Lila , Jin Fang , Johnson Aaron , Ayasoufi Katayoun
Published: November 2024

Peripheral nervous system innervation of various organs has been associated with the development and maintenance of the immune system. Interestingly, various neurological insults including brain tumors, intracranial metastases, and neurotropic infections have been shown to have profound impacts on peripheral immunity, including thymic size and cellularity. Therefore, we sought to determine whether the thymus was directly innervated. Using flow cytometry and confocal microscopy, we identified a population of cells within the thymus that contained both neuronal markers and morphology. Subsets expressed tyrosine hydroxylase, BIII tubulin and/or NeuN, distinguishing them from thymic epithelial cells. These cells do not express AIRE, further highlighting their neuronal identity. Interestingly, a subset of these neurons also express MCH-II, implying a potential role in T cell selection. Neuronal cell bodies and/or axonal processes were identified within the thymus. Using retrograde rabies virus-derived neurotracers, we confirmed axonal connections and the identity of thymic neurons. Finally, both neuronal and epithelial cells were absent in RAG-deficient mice, suggesting a shared differentiation or survival pathway. Both populations could be restored by introducing healthy bone marrow. To summarize, we describe a novel population of neuronal cells within the thymus that interact with developing thymocytes to establish optimal thymic architecture.
Self-supervised deep learning enables label-free, high-dimensional morphology profiling of immune cell types
Authors: Balakrishnan Michelle , Boutet Stephane , Saini Kiran , Zhang Senzeyu , Wong-Thai Amy , Lu Vivian , Carelli Ryan , Jacobs Kevin , Jovic Andreja , Mavropoulos Anastasia , Masaeli Maddison
Published: November 2024

Immunophenotyping & detecting cell subsets are key parts of immunology research. To determine whether morphology as a quantitative, high-dimensional analyte (morpholome) can phenotype and distinguish among cell types found in PBMCs more comprehensively & without labels we used the REM-I platform to assess patient-derived PBMC samples & purified immune cell subsets.

The REM-I platform combines high-speed imaging in flow on the REM-I instrument, and deep learning & computer vision in the Human Foundation Model (HFM). The HFM produces 115 embeddings that are reproducible, quantitative descriptions representing the morpholome of each cell in a sample that can be analyzed through interactive UMAPs, population analytics, & image visualizations.

Morpholomic analysis of patient-derived PBMC samples and purified immune cell subsets including CD4+ T cells, CD8+ T cells, CD14+ monocytes, CD19+ B cells, CD38+ plasma cells, CD56+ NK cells, and macrophages, showed profiled cell subsets in distinct locations on the associated UMAP, indicating cell types are separable by morpholomics. Leiden clustering segregated cell groups by morphological features, with some cell subsets sharing common leiden clusters. Purified cell types derived from different donors consistently localized to the same high-density UMAP regions, indicating that diverse morphotypes are reproducible across different donors, & suggesting a link between morphological profile and cell function.
Keratin 17- and PKCα-dependent amplification of neutrophil influx after repeated stress to skin
Authors: Xu Yang , Cohen Erez , Johnson Craig , Parent Carole , Coulombe Pierre
Published: November 2024

Neutrophils contribute to the pathogenesis of chronic inflammatory skin diseases. Little is known about the source and identity of the signals mediating their recruitment in inflamed skin. We used the phorbol ester TPA and UVB, alone or in combination, to induce sterile inflammation in mouse skin and assess whether keratinocyte-derived signals impact neutrophil recruitment. A single TPA treatment results in a neutrophil influx in the dermis that peaks at 12h and resolves within 24h. A second TPA treatment or a UVB challenge, when applied at 24h but not 48h later, accelerates, amplifies, and prolongs neutrophil infiltration. This transient amplification response (TAR) is mediated by local signals in inflamed skin, can be recapitulated in ex vivo culture, and involves the K17-dependent sustainment of Protein Kinase C α (PKCα) activity and release of neutrophil chemoattractants by stressed keratinocytes. We show that K17 binds RACK1, an essential scaffolding protein for PKCα activity. Analyses of RNAseq data reveal the presence of a transcriptomic signature consistent with TAR and PKCα activation in chronic inflammatory skin diseases including psoriasis, hidradenitis suppurativa, and atopic dermatitis. These findings uncover a novel, transient, and keratin-dependent mechanism that amplifies neutrophil recruitment to the skin under stress, with direct implications for inflammatory skin disorders.
Surface CD6 regulates T cell adhesion and migration through direct ligand interaction and stabilization of VLA4
Authors: Ampudia Jeanette , Marrocco Valeria , Ritter Madison , Connelly Stephen , Ng Cherie
Published: November 2024

The CD6-ALCAM pathway is an important regulator of T cell activity and trafficking. However, its precise role in modulating T-cell adhesion and migration during inflammation is unclear. To investigate its role in adhesion and transendothelial migration, we used itolizumab, an anti-CD6 mAb that can induce cleavage of CD6 from the T cell surface when in the presence of monocytes. PBMCs were stimulated overnight or left unstimulated prior to treatment with itolizumab to generate CD6low T cells. Cells were then tested in standard adhesion and migration assays.

The CD6 low T cells had ~ 50% decreased adherence to HUVEC cells compared to the control T cells (CD6high); similar results were observed by blocking ALCAM on HUVEC. Modulating CD6 on T cells or blocking ALCAM on the HUVEC also translated to a ~3-fold decrease in migration of Teff cells.

Loss of CD6 was associated with reduced protein levels of VLA4, but not of other integrins, under both stimulated (~70%) and non-stimulated conditions (~40%). Antibody blockade of CD6 without cleavage did not affect VLA4 levels, but still decreased migration of unstimulated T cells though less so than in stimulated T cells, where VLA4 reduction is required to modulate migration.

Our data demonstrate that CD6 contributes to cell adhesion/migration both through direct ligand interactions with ALCAM, and through the stabilization of cell membrane VLA4 and that these functions can be abrogated by modulation of cell surface levels of CD6.
Compromised antibody responses and memory lymphocyte formation to COVID-19 mRNA vaccination in children with acute lymphoblastic leukemia undergoing maintenance chemotherapy
Authors: Chang Pei-De , Shieh Chi-Chang
Published: November 2024

COVID-19 mRNA vaccines, with 95% efficacy in the general population, have been shown to exhibit suboptimal effectiveness in hematological malignancy patients. Our previous published study revealed that COVID-19 vaccine responses were related to innate immune responses. In this study, we investigated the innate and adaptive immune responses to COVID-19 mRNA vaccines in children with acute lymphoblastic leukemia (ALL). We longitudinally investigated the innate immune responses to PRR-agonists stimulation and after vaccine injection, along with vaccine-induced anti-S antibodies, S-specific memory B and T cell responses in 13 ALL children who completed or were undergoing maintenance chemotherapy and 24 healthy controls. We found that both ALL groups had impaired dsDNA-induced cytotoxic NK cell activation while patients in maintenance therapy also showed decreased dsRNA-induced activation in those cells. After vaccination, ALL children with completed therapy demonstrated similar S-specific antibody responses and memory B cells as healthy controls. In patients with maintenance therapy, these responses were notably reduced, especially cross-protective antibodies. Both ALL groups exhibited lower S-specific CD8+ T cells, which correlated with weaker TLR9 responses. In conclusion, we discovered the compromised vaccine responses in ALL children, especially those in maintenance chemotherapy, highlighting the importance of improved COVID-19 vaccines for this special group of patients.
Development of swine Fc-conjugated serotype O foot-and-mouth disease virus-like particles and immunological evaluation in swine
Authors: Shin Hyun-Jin , PARK JAE-YEON
Published: November 2024

The outbreak of foot-and-mouth disease (FMD) has been declining as a result of efforts by many advanced countries to eradicate it. However, new vaccine types are being developed because FMD is still a problem in several countries, and vaccinations have safety concerns. In this study, we developed a new FMD vaccine using virus-like particles (VLPs) harboring the immunoglobulin G Fc molecule on their surface. Based on the results of vaccination studies in swine, we confirmed its high efficacy. In particular, we confirmed strong immune boosting by the addition of the swine IgG (sFc) region. The FMDV VLP-sFc vaccine produced higher amounts of Th1 cytokines (IFN-γ, IL-12, and TNF) and Th2 cytokines (IL-4) than the commercial vaccine. More importantly, FMDV VLP-sFc induced significantly higher levels of neutralizing antibodies. Taken together, our newly developed FMDV vaccine, FMDV VLP-sFc, may be a great new and effective FMDV vaccine.
mRNA delivery of epitope-specific monoclonal antibodies for treatment of viral infection
Authors: Hazell Nicholas , Hu Haitao , Bonam Srinivasa Reddy , Hajnik Renee , Nguyen Tina
Published: November 2024

The mRNA-lipid nanoparticle (mRNA-LNP) platform is versatile for development of vaccines and therapeutics, specifically for monoclonal antibody (mAb) treatment of viral disease. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected more than 760 million worldwide and contributes to nearly 7 million associated fatalities. As vaccines protect those immune-competent, the elderly and immune-compromised share increased risk for development of severe lung disease. In addition, virus evolution leads to novel variants that pose an enhanced threat to the aforementioned subpopulations, highlighting a need for therapeutics that can rapidly be developed to prevent severe disease in these high-risk populations. mAbs that neutralize are effective for treating SARS-CoV-2 infections with a number of mAbs targeting SARS-CoV-2 identified. We generated mRNAs expressing two different mAbs, LY-CoV1404 (mRNA-1404) and 76E1 (mRNA-76E1), that target different SARS-CoV-2 epitopes and formulated them in LNPs. In vitro analysis support both mRNAs can produce and secrete mAbs with epitope specificity in cells following transfection and in vivo studies exhibit durable antibody expression. Following single mRNA-LNP administration (5ug and 15ug dose via intramuscular or subcutaneous route), serum antibodies remained detectable 5 weeks out. Our data indicate that mRNA is a promising platform for the delivery of therapeutic mAbs to treat human disease such as viral infections.
A combination of TLR9 and STING agonists favors antigen cross-presentation and promotes Th1 immune response against influenza virus
Authors: Thomas Sanya , Pak Jensen , Doss-Gollin Simon , Ryff Kevin , Beijnen Elizabeth , Pedersen Gabriel , Christensen Dennis , Levy Ofer , van Haren Simon
Published: November 2024

Adjuvants can boost the type, magnitude, breadth, and durability of vaccine responses. We have previously demonstrated that combining adjuvants through rational design can overcome age-specific impairments in inducing Th1 immunity, trigger antigen cross-presentation, and enable CD8-driven protective immunity of protein-based vaccine formulations. Here, we screened 78 adjuvant combinations to explore their molecular mechanism of action through human in vitro modeling and identified combinations that synergistically promote the expansion of antigen-specific CD4+ cells, induce cross-presentation on MHC class I resulting in activation of antigen-specific CD8+ cells, and redirect the type of immune response to favor the production of Th1-promoting cytokines. The combination of CpG (TLR9 agonist) and 2,3-cGAMP (STING agonist) promoted influenza-specific CD4+ and CD8+ T cell activation and selectively favored the secretion of Th1-polarizing cytokines TNF and IL-12p70. Activation of T-bet-expressing Th1 cells was also observed with this adjuvant combination. We also demonstrate the phenotypic reprogramming towards conventional type 1 dendritic cells by CpG + 2,3-cGAMP, and the ability of this adjuvant combination to induce similar immune responses in immune cells from newborns and older adults. Identification of the adjuvant combination CpG + 2,3-cGAMP may prove key to the future development of vaccines against intracellular pathogens such as respiratory viruses.
Cytosolic nucleic acid sensors stimulate protective bone cell responses to Staphylococcus aureus
Authors: Johnson Morgan , Sipprell Sophie , Suptela Samantha , Mills Erin , Marriott Ian
Published: November 2024

Staphylococcus aureus is the causative agent of osteomyelitis, an inflammatory infection of the bone. Despite available treatments, S. aureus invasion of resident bone cells limits antibiotic effectiveness and provides a protective niche for viable bacteria contributing to recurrent and persistent infections. Previously, we have demonstrated a significant increase in proinflammatory cytokines, chemokines, and osteoclastogenic factors in an in vivo model of localized Staphylococcal osteomyelitis. Interestingly, we also observed a significant production of type I IFNs in infected femurs compared to the contralateral control. It is now appreciated that resident bone cells express a variety of pattern recognition receptors for identification of pathogens and stimulation of immune responses. Here, we demonstrate that S. aureus infected osteoblasts produce significant increases in both mRNA encoding type I IFNs and protein. Our data demonstrates that cytosolic nucleic acids sensors, including the RNA sensor RIG-I and the DNA sensor cGAS, are upregulated by osteoblasts and initiate osteoblast responses to S. aureus challenge. Notably, S. aureus viability is necessary for stimulation of osteoblast production of type I IFNs. Furthermore, our data indicate that IFN-β serves to restrict intracellular S. aureus burden. Our ongoing studies seek to investigate the therapeutic potential of augmenting these protective bone cell responses in a murine model of Staphylococcal osteomyelitis.
Recall of cross-reactive memory B cells enhances antibody durability and breadth against SARS-CoV-2 variants
Authors: Chen Yuezhou , Wang Youyi , Hu Yuanhao , Gong Nuo , Li Yunfeng , Huang Jiayi , He Jiaxin , Huang Shun
Published: November 2024

Long-lasting antibody production with broad recognition breadth is crucial to combat with many pathogens. After infection or vaccination, B cell memory is preserved via two key elements: memory B cells, which can be efficiently recalled during subsequent encounters, and long-lived plasma cells, which continue to produce antibodies over the long term. It's critical to understand antibody durability and how memory can identify different viral variants following iterative exposures. We examined the recognition of SARS-CoV-2 variants, the dynamics of memory B cells, and the secretion of antibodies over time after recurrent infections and vaccinations. Within unvaccinated individuals who recovered from COVID, enhanced antibody stability over time was observed within a subgroup of individuals who recovered more quickly from COVID, had greater percentage of spike specific memory circulating T follicular helper (cTfh), and harbored significantly more memory B cells cross-reactive to endemic coronaviruses early after infection. These cross-reactive clones map to the conserved S2 region of SARS-CoV-2 spike with higher somatic hypermutation levels and affinity. Following SARS-CoV-2 inactivated vaccination, frequency of cross-reactive memory B cells after BF.7 infection links to antibody longevity and neutralization breadth to SARS-CoV-2 variants. We conclude that memory B cell recall shapes functional cross-variant antibody repertoire composition and longevity.
The inhibitory role of nitric oxide in encephalomyocarditis viral replication in pancreatic β-cells.
Authors: Gehant Alyssa , Stafford Joshua , Corbett John
Published: November 2024

During type 1 diabetes (T1D) onset, insulin-secreting β-cells are selectively destroyed by T-cells. In addition to known genetic factors, viral infection (specifically by members of the picornavirus family) is an environmental factor that has been implicated in T1D onset. Viral infection stimulates macrophage release of IL-1, and in islets, stimulates the production of micromolar levels of nitric oxide (NO) by β-cells. NO impairs β-cell function via inhibition of mitochondrial oxidative metabolism, resulting in depletion of ATP. Since β-cells have a limited capacity for self-renewal, and are essential for organismal survival, we hypothesize that the cytokine-mediated production of NO by β-cells functions in a protective manner during a viral infection. In support of this hypothesis, we’ve shown that NO inhibits encephalomyocarditis viral (EMCV) replication and mediated lysis in a β-cell selective manner. EMCV is an inflammatory, mouse-tropic virus that induces diabetes in several mouse models. In this study, we have identified inositol hexakisphosphate kinase-1 (IP6K1, KmATP=1 mM) as an indirect target of NO (via the loss of ATP) that is responsible for the inhibition of EMCV replication in β-cells. We show that inhibition of IP6K1 attenuates EMCV replication and mediated β-cell lysis in a manner comparable to the actions of NO. Current studies are focused on the mechanism by which inhibition of IP6K activity attenuates EMCV replication and EMCV-mediated lysis in β-cells.
The CD40/CD40L dyad, an immune checkpoint regulator, plays a protective role against Mouse hepatitis virus-induced neuroinflammatory demyelination
Authors: Das Sarma Jayasri , Hazra Bishal
Published: November 2024

Murine ꞵ-coronavirus, RSA59, infection results in meningoencephalitis, demyelination, and axonal loss. CD40-CD40L dyad serves as an important immune checkpoint regulator during this disease pathobiology. C57BL/6 WT, CD4-/-, CD40L-/- and CD40-/- mice were intracranially inoculated with RSA59 to study the acute phase neuroinflammation and chronic phase demyelination. Studies suggested that CD4+ T cells play a protective role against RSA59-induced demyelination because, in the absence of CD4 gene, mice were more susceptible to acute poliomyelitis and chronic demyelination with axonal bulbar vacuolation. CD40L expressed by CD4+ T cells is crucial for host immunity against RSA59. Without CD40L in activated CD4+ T cells, RSA59-infected mice experience increased disease severity, dampened microglia/macrophage activation, reduced T cell infiltration in the CNS, and impaired T cell priming in the cervical lymph node. In the chronic phase, extensive CNS virus replication leads to severe demyelination and axonopathy due to altered microglia/macrophage populations caused by the absence of the CD40L gene. CD40-deficiency revealed increased acute phase encephalitis in the RSA59-infected groups due to increased infiltrating neutrophils, leading to heightened bystander neuroinflammation. Hence, it is evident that the RSA59-induced demyelination model elucidates an extensively robust innate immune system upon which adaptive immunity is built during the chronic stage of infection.
NLRP3 Cys126 palmitoylation by ZDHHC7 Promotes Inflammasome Activation
Authors: Yu Tao
Published: November 2024

Macrophages could response to pathogen and damage-associated molecular patterns through NLRP3-mediated inflammasome. Hyperactivation of NLRP3 is associated with human chronic inflammatory diseases. Therefore, accurate regulation of NLRP3 activation is important. Recent works show that NLRP3 is localized on trans-Golgi network (TGN), which is crucial for NLRP3 activation and consequently ASC recruitment. However, what regulates NLRP3 TGN localization is unknown. Here, we demonstrated that NLRP3 is palmitoylated on Cys126 by palmitoyl-acyltransferase ZDHHC7 in macrophages, which promotes resting NLRP3 localization to TGN and activated NLRP3 localization on dispersed TGN. As a result, ZDHHC7-mediated NLRP3 Cys126 palmitoylation is critical for recruitment and oligomerization of ASC after NLRP3 activation. Perturbating NLRP3 palmitoylation by ZDHHC7 knockout, pharmaceutically inhibition, or modification site mutation, will diminish NLRP3 activation and consequential Caspase-1/GSDMD cleavage, and cytokines secretion in macrophages. Furthermore, NLRP3 palmitoylation on Cys126 is vital for inflammasome activation in vivo, including endotoxic shock and peritonitis mouse models. Notably, this palmitoylation-regulated NLRP3 activation also contributes to multiple sclerosis development in mice. Therefore, our study provides new insight into NLRP3 regulation and suggests targeting ZDHHC7 or NLRP3 Cys126 could be a potential therapeutic strategy to treat human NLRP3-related disorders.
The transcription factor Runx3 promotes diversity in effector cell precursors that pattern the memory CD8 T cell compartment
Authors: Albao Dominic , Diao Huitian , Pipkin Matthew
Published: November 2024

Central memory (TCM) CD8 T cells maintain the regenerative capacity of long-term adaptive immunity to intracellular pathogens, but recent studies indicate that TCM cells are heterogeneous. Previous analyses of the transcription factor Runx3 has shown it has pleiotropic effects on memory CD8 T cell development, including potential diversity among TCM cells: Runx3-deficient cells preferentially form Tcf7hi CD62Lhi TCM-like cells but accumulate inefficiently, whereas enforced Runx3 expression promotes increased numbers of CD62Lhi TCM – like cells and more efficiently form memory precursors upon secondary infection. Using single cell RNA sequencing during acute LCMV infection we identified two distinct effector cell states enriched with TCM gene expression that differentially correlated with Runx3-dependent transcription (Runx3 signature). One state exhibited a stem-like signature and a low Runx3 signature score, whereas the other was less stem-like and exhibited a higher Runx3 signature. Retroviral expression indicated that wildtype Runx3 promoted formation of CX3CR1int peripheral memory-like and TCM-like cells, and repressed effector memory (TEM) cell formation, whereas expression of Runx3 lacking the C-terminal VWRPY motif caused divergence into a TEM-like state. These results suggest that the dynamic activity of Runx3 and the potential interactors with its specific domains mediate complex regulation that patterns antiviral memory CD8 T cell development.
SATB1 is essential in regulating the stemness of antigen-specific CD8+ T cells
Authors: Lin Siying , Cui Weiguo
Published: November 2024

In chronic infection and cancer, a subset of self-renewing, antigen-specific CD8+ T cells, termed progenitors, is crucial for long-term immunity and immunotherapy effectiveness. The mechanisms governing their stemness, however, are not fully understood. Our single-cell RNA + ATAC sequencing analysis revealed that SATB1 (special AT-rich sequence-binding protein) is exclusively expressed in the progenitor CD8+ T cells during chronic infection. Known for its key role in chromatin organization during T-cell development, SATB1’s unique expression in the progenitor CD8+ T cell led us to hypothesize that it plays a pivotal role in regulating the stemness of the antigen-specific CD8+ T cells. Using the lymphocytic choriomeningitis virus (LCMV) Clone 13 mouse model of chronic infection, the SATB1 gene was deleted in CD8+ T cells via CRISPR/ RNP technology. At day 21 post-infection, the SATB1 deletion resulted in a reduction of virus-specific progenitor CD8+ T cells and a decrease of TCF1 expression. The transcriptomic and epigenomic studies further revealed that SATB1 plays a key role in regulating the chromatin accessibility and transcriptional activities of stemness-associated genes, such as Tcf7, Bach2, and Myb. Overall, our results underscore the critical role of SATB1 in maintaining the transcriptional integrity of progenitor CD8+ T cells in response to antigen activation, which sheds light on the genetic pathways that regulate the stemness of antigen-specific CD8+ T cells.
Age-related dynamics of CD8+ regulatory T cells: Implications of NKG2D in Immune Aging
Authors: Srinivasan Saranya , Ma Chaoyu , Wang Liwen , Mishra Shruti , Fan Kenneth , Im John , Segura Courtney , Zhang Nu
Published: November 2024

Aging is a complex biological process that impacts various physiological functions, including the immune system. Our study investigated the impact of aging on CD8+ regulatory T cells(CD8+Treg), which showed a pattern of initial increase up to 12 months, then a decline by 24 months in a C57BL/6 mouse model, unlike memory T cells, which consistently increase with age. Functionally, CD8+Treg, irrespective of age, did not produce cytokines in response to TCR stimulation, highlighting a stark functional contrast with memory cells. However, upon IL-15 stimulation, CD8+Tregs, unlike their memory cell counterparts, demonstrated enhanced cytokine production. Transcriptomic analysis revealed an abundance of Klrk1 (killer cell lectin-like receptor subfamily K member 1)gene transcripts in aging CD8+Tregs. Klrk1 gene encodes Natural Killer Group2 Member D (NKG2D), an activating receptor NK cells express. This upregulation was specific to CD8+CD122hiLy49+ and absent in CD8+CD122hiLy49-. To explore this correlation further, we utilized a Klrk1-/- mouse and observed an increased CD8+Treg population, suggesting a potential negative regulatory role of NKG2D in CD8+Treg homeostasis. Transcriptome analysis of Klrk1-/- CD8+Tregs showed enrichment of aging-related gene signatures, further corroborating the link between NKG2D and the aging process. Our findings suggest NKG2D's involvement in aging and immune dysfunction, providing avenues for intervention in the elderly.
Development of single-cell ROS regulome profiles of CD8+ T cells
Authors: WANG YI-CHUAN
Published: November 2024

Cells under oxidative stress regulate their functions, but current methods for detecting this stress mostly use mass spectrometry or fluorescent probes sensitive to reactive oxygen species (ROS). While valuable, these methods can't simultaneously characterize specific ROS-mediated pathways at the single-cell level. Thus, we developed an approach to characterize the ROS regulome of single-cells together with their phenotypic identity, termed single-cell ROS regulome profiling (scROP). To accomplish this, we first assessed over 100 commercially available antibodies by fluorescent cell barcoding screening and then selected a subset of ROS antibodies (n = 25). We combine our method with in vitro-activated OTI-CD8+ T cells to quantify the proteins that regulate the activity of the ROS pathway, it provides a more detailed sequential progression of how ROS regulome response through oxidative and redox pathways in different CD8+ T cell states that lead to exhaustion. Moreover, our results show that T cell exhaustion rates differ under hypoxic conditions or in the presence of antioxidants. We confirmed our findings using clinical samples, such as blood from hemodialysis patients, CD8+ T cells from CAR-T patients, and various tumor sections from hepatic carcinoma patients. Overall, our novel single-cell ROS regulome profiling approach revealed a comprehensive molecular roadmap of CD8+ T cells in response to sustained stimulations.
Human Immune repertoire and gene expression in pediatric and adult SARS-CoV-2 infected patients using single cell multi-omics approaches
Authors: ji xuhuai , Yonker Lael , Miranda Molly , Kinane Thomas Bernard , Chang Matthew , Suresh Sundari , Kiany Samaneh , Weinacht Katja , Maecker Holden
Published: November 2024

SARS-CoV-2 represents a global health crisis, yet major knowledge gaps remain about the human immune response against the virus infection and related treatment in both pediatric and adult patients. Immune repertoires (TCR and BCR) and related gene expression at massive single cell level play a crucial role in early-stage infection and late virus clearance. By using single cell multi-omics approaches, we analyzed immune repertoire and gene expression in 9 COVID-19 patients and 3 controls. We used single-cell mRNAseq and observed increased expression of inflammatory cytokines and transcription factors such as CXCL1, RNASE2, EGR1, CXCL2, and CXCL8 in either adult or pediatric COVID-19 infected patients compared to controls. Ten surface proteins such as CD62L, CD16, CD20, CD56, CD14, and CD45RA were quantitated by AbSeq. CD14+ cells were more prevalent in adult COVID-19 patients and CD3+ T cells were less prevalent. We conducted comparative analyses of TCR and BCR repertoire using single-cell V (D)J sequencing approach. The clonotypes were defined by using either combined alpha and beta chains or beta chains only. Distinct clonotypes were identified in both pediatric and adult COVID-19 patients. We did not see any significant clonotype expansion in this sample set yet but further analysis is warranted. The combination with the highest frequency in COVID-19 was TRAV29-TRBV27. Our study provides novel insights and bioinformatic analysis tools on immune repertoire in COVID-19.