bluuebunny/crossref_metadata_2025_split huggingface dataset:
A novel mutation in IRAK2 results in immune dysregulation in systemic juvenile idiopathic arthritis (sJIA)
Authors: Boyd Brooke , Platukus Alana , Schmitz Elizabeth , Nakano Hiroto , Dell'Orso Stefania , Naz Faiza , Deng Zuoming , Cruz Anthony , Ombrello Michael 
Published: November 2024

Abstract
Systemic juvenile idiopathic arthritis (sJIA) is a severe inflammatory disease marked by innate immune activation with potentially fatal complications. Using trio exome sequencing, we identified a novel homozygous mutation in interleukin (IL)-1 Receptor Associated Kinase 2 (IRAK2) in a child with sJIA complicated by lung disease and hyperinflammation. IRAK2 is a serine/threonine kinase involved in IL-1 receptor and toll-like receptor signaling, leading to expression of pro-inflammatory genes. The patient displayed persistent elevation of IL-6, IL-8, and IL-18 and a strong interferon (IFN) stimulated gene (ISG) signature. To determine how the IRAK2 mutation contributes to the patient’s immunopathology, we used gene editing to create THP-1 cells homozygous for the mutation (THP-IRAK). Following 24 hours of lipopolysaccharide (LPS) treatment, RNA sequencing revealed that THP-IRAK cells display higher expression of IL-6, IL-18, and ISGs than wild type THP-1 cells (THP-wt). Gene set enrichment analysis of MSigDB hallmark sets identified LPS-induced upregulation of IFN signaling and cell cycle regulation related sets in THP-IRAK cells relative to THP-wt cells. Finally, LPS treatment led to higher secretion of IL-8 in THP-IRAK cells than in THP-wt cells. Together, these in vitro results recapitulate the pattern of inflammation observed clinically, which suggests that the IRAK2 mutation contributes to the pathophysiology of sJIA and nominates IRAK2 as a therapeutic target in sJIA.
Controlled Reduction of Reactogenicity in Vaccines - Designing New Molecules to Alter Innate Signal Processing
Authors: Esser-Kahn Aaron , Kim Jeremiah , Roman Maria , Snyder Scott 
Published: November 2024

Abstract
Vaccines are a critical tool in preventing and managing many diseases, but their efficacy can be improved by enhancing their immunogenicity and lowering their reactogenicity. We have developed a series of molecules that directly reduce correlates of reactogenicity while also improving protection. We present data on the application of these immunomodulator towards a CpG adjuvanted subunit SARS- CoV-2 vaccine in both mice and Syrian hamsters. Immunomodulator-containing vaccines increased neutralizing antibody and T-cell responses in mice. In a challenge model in hamsters, these vaccines reduced weight loss, viral load, and tissue damage. These findings demonstrate the possible improvement of current and future vaccines using immunomodulators. We also present a series of modulators that reduce and direct inflammatory responses in a controlled fashion for use in influenza, hepatitis, typhoid, and many other vaccine applications.
Production of a covalent conjugate vaccine using cell-free protein expression (XpressCF™) and click chemistry
Authors: Rozzelle James , Poston Taylor , Arroyo Daisy , Bruno Antonio , Wu Manhong , Esser-Kahn Aaron , Darville Toni , Fairman Jeff 
Published: November 2024

Abstract
Chlamydia trachomatis is the most common bacterial sexually transmitted infection globally and a vaccine is urgently needed. To address this, we determined that Chlamydia protease-like activity factor (CPAF) is the most immunoprevalent and immunodominant antigen for CD4 T cells and B cells in Chlamydia exposed women and mice. CPAF is a 70-kDa serine protease that is secreted into the host cell cytoplasm and subsequently released into the extracellular milieu where it paralyzes neutrophils to evade phagocytosis. CPAF contains a fold similar to other known serine proteases and is processed into C- and N-terminal fragments that heterodimerize. To improve and facilitate vaccine scale-up and immunogenicity, we developed CPAF variants incorporating non-native amino acids at specific sites using the proprietary cell free expression platform XpressCF™. These CPAF variants were covalently conjugated to DBCO-derivatized adjuvants using click chemistry to make novel antigen-adjuvant conjugate vaccines. Site-specific conjugation enables important T- and B-cell epitopes to be avoided for better immune recognition and provides a method to incorporate a specific number of conjugated adjuvants. These conjugated vaccines provide a means to simultaneously deliver antigen and adjuvant and potentially reduce the amount of adjuvant required for vaccine efficacy leading to fewer side effects and dose sparing.
Repeated allergen consumption impairs intestinal barriers and promotes neurobehavioral pathologies in mice with subclinical cow’s milk allergy
Authors: Ekanayake Dilini , Setness Rylan , Nagamoto-Combs Kumi 
Published: November 2024

Abstract
Cow’s milk and other food allergens are often implicated in behavioral problems in sensitized but tolerant individuals. While the mechanisms are still unclear, we previously showed that male C57BL/6J mice with subclinical cow’s milk allergy (CMA) exhibited depression-like behavior associated with brain neuroinflammation and cortical demyelination after consuming a whey protein-containing diet (WP diet) for 2 weeks. Recently, we also found that adoptively transferred peripheral leukocytes from CMA mice migrated to the dura mater of naïve recipient mice during the duration of the 2-week WP diet. Hypothesizing that consumption of food allergens by subclinically sensitized individuals would cause intestinal damage and promote the trafficking of activated peripheral leukocytes to the dura to adversely affect the brain, we assessed neurobehavioral and intestinal pathologies CMA mice that sensitized to β-lactoglobulin (Bos d 5) and exposed to WP diet for 2 weeks. Behavioral tests indicated that the exploratory activity and short-term memory of CMA mice were significantly lower than controls. Cortical demyelination, microgliosis, and axonal damage were detected in the brain as previously reported. Notably, immunoreactivity for a tight junction protein, claudin 5, was visibly lower in the intestines of CMA mice. Our results suggested that repeated allergen consumption could trigger immune responses by increasing gut permeability in susceptible individuals.
The calpain/ABCA1 pathway drives mast cell- initiated vascular leakage and inflammation in the skin.
Authors: Kohl Lisa Marie , Hay Ahnika , Gupta Vinay , Williams Li'an , Bharadwaj Shreya , Donahue McKenna , Omar Lea , Cattley Richard , Sumpter Tina 
Published: November 2024

Abstract
Cutaneous mast cells (MCs) initiate local and systemic immune responses. To reach this end, activated MCs immediately release an arsenal of pro-inflammatory proteases. In this study, we evaluated the role of the cysteine protease, calpain in MC function. Calpain activity and protein were confirmed in the supernatants of MCs activated via either the FceR or the MrgprB2. In vivo, extracellular calpain induces vascular leakage in a MC- dependent manner. We then investigated the mechanism of calpain release. In vitro and in vivo studies suggest that the cholesterol transporter, the ATP- binding cassette A1 transporter (ABCA1) externalizes calpain. To investigate the role of the ABCA1 in MC biology, we generated mice lacking the Abca1 specifically on connective tissue MCs (Mcpt5-Cre+Abca1fl/fl mice). In vitro, Abca1-deficient MCs accumulate intracellular cholesterol, have reduced calpain release and reduced pro-inflammatory cytokines levels. In vivo, in the MC903-induced atopic dermatitis model, Mcpt5-Cre+Abca1fl/fl mice have reduced inflammation and a fewer MCs in affected skin compared to littermate controls. Corroborating this finding, global ABCA1 blockade with FDA-approved glyburide similarly reduced inflammation induced by MC903. Collectively, these data suggest that targeting the calpain/ABCA1 pathway may open new therapeutic avenues for MC -mediated inflammation in the skin.
GEMINI pipeline: Fecal microbiota and Immune fitness evaluation in healthy, elderly and centenarians populations
Authors: Levast Benoit , Beitz Benoît , Mouharib Manal , Ania Maryne , Bellais Samuel , Ben Abdallah Bouthaina , Koenig Marie , Welsh Jérémy , Beloeil Laurent , Louis Karen , Guyard Cyril 
Published: November 2024

Abstract
The symbiosis between microbiome and its host creates a homeostasic crosstalk regulating the host immune system. Host microbiome richness and diversity are linked with a wide range of diseases and treatment efficacies such as vaccination or immunotherapies.
The project includes three different cohorts of volunteers: healthy volunteers (18 and 55 years old) – considered as a reference group (n=51); elderly population between 50 and 75 years old - considered as the population of interest for seasonal vaccinations (n=60); and centenarians’ population (&gt; 95 years old) – that may present a gut microbiota and immune system with specific properties partly supporting their exceptional longevity in relatively good health.
By integrating high throughput anoxic culturomics, host/microbiota interaction functional analyses, with metabolism and host immune response analytical capacities, BIOASTER has established a multidisciplinary platform for the development of microbiome derived products.
Here we report the characterization of human donors “immune fitness” data through the analyses of plasma cytokines profiles in whole blood stimulation assays using TruCulture, paired to feces analyses. Our results will guide our strategy for gut microbiome bacterial isolations.
The screening of specific microbiome strains will lead to their genetic identification, their functional characterization and the development of next-generation probiotics with clinical related immuno-modulatory properties.
Deciphering TCR-pMHC Interactions in Human Memory T Cells: Biophysical Insights into SARS-CoV-2 Antigen Recognition
Authors: Chen Xiangcheng 
Published: November 2024

Abstract
Precise antigen recognition by T cell receptors (TCRs) is key for initiating cellular immune responses. This study investigates the biophysical characteristics of TCR-antigen interactions, emphasizing 2-dimentioanl (2D) TCR affinity and catch bond dynamics in human memory T cell subsets, particularly in their response to specific SARS-CoV-2 epitopes.
Utilizing DNA-barcoded pMHC tetramers, our research profiles TCRs interacting with selected SARS-CoV-2 epitopes, identifying antigen-specific memory T cells in lung tissue, peripheral blood mononuclear cells (PBMCs), and antigen-specific precursor T cells from the spleen. Through micropipette adhesion assays and biomembrane force probe, we quantified 2D TCR affinity and examined catch bond formation dynamics at the single-cell level.
Our findings indicate a broader range of 2D TCR affinities in spleen precursor T cells compared to memory T cell subsets in tissue and blood. Notably, tissue-resident and circulating memory T cells exhibit distinct 2D TCR affinity and catch bond properties.
The differential biophysical properties, particularly in 2D affinity and catch bond force lifespan, underscore the unique role of tissue-resident memory T cells, influencing memory formation and tissue-specific immune responses. This detailed characterization of TCR-pMHC interactions in human tissue-resident memory T cells contributes significantly to our understanding of memory generation during viral infections.
Sialic acids from the Group B Streptococcus (GBS) capsule dampen mast cell activation through Siglec-9
Authors: Miralda Molina Irina , Orvis Austyn , Coleman Michelle , Brokaw Alyssa , Lewis Amanda , Rajagopal Lakshmi , Piliponsky Adrian 
Published: November 2024

Abstract
GBS is a leading cause of bacterial infection in newborns, adult diabetics, pregnant women, and the elderly. A key virulence factor of GBS is its surface capsular polysaccharide, which displays terminal sialic acids that suppress host innate immune responses by engaging host inhibitory receptors like Sialic acid-binding immunoglobin-like lectin- 9 (Siglec-9). We reported that human mast cells (MC) express Siglec-9 and that engaging it with native ligands or antibodies results in reduced mast cell activation in vitro. We hypothesized that capsular sialic acids engage Siglec-9 to impair MC ability to initiate the host response. Mice expressing human Siglec -7 and -9 were infected with WT GBS or a sialic acid deficient strain, ΔcpsK, and the ΔcpsK infected mice showed reduced bacterial burden and less dissemination. Using bone marrow-cultured MC from a transgenic mouse expressing Siglec-9, we confirmed that GBS interacts with MC Siglec-9, which inhibited the release of the pro-inflammatory cytokine IL-6. Challenge of primary human MC with WT, ΔcpsK, or ΔneuA, another sialic acid deficient GBS strain, showed that MC interact more with ΔcpsK and ΔneuA GBS, and that MC kill sialic acid deficient GBS. MC challenged with ΔcpsK or ΔneuA GBS also produce more IL-8 and undergo cell death more than WT infected MC. Together, this data shows that sialylated GBS capsule engages MC Siglec-9 to dampen activation, which reduces the immune response and benefits the pathogen.
A humanized ACE2 mouse model recapitulating age- and sex-dependent immunopathogenesis of COVID-19
Authors: Park Uni , Lee Jae Hoon , Kim Uijin , Kim Yuri , Jeon Kyeongseok , Kang Ju-il , Kim Hyeran , Oh Song Hyuk , Kang Jun-Gu , Han Seung Ro , Yoon Sang Ho , Jin Liyuan , Zhang Yinhua , Kim Myoung-Hwan , Lee Myung-Shin , Cho Hyun-Soo , Lee Han-Woong 
Published: November 2024

Abstract
The COVID-19 pandemic underscores the critical requirement for robust animal models to elucidate the immunological and pathological repercussions of emerging pathogens. In this study, we introduce a novel humanized ACE2 mouse model (hACE2-E1/2/8) developed through precise CRISPR-based substitution of nine human ACE2 amino acids (N24Q, N30D, N31K, Q34H, T79L, S82M, F83Y, S84P, and H353K). This model maintains mouse ACE2 physiological function, exhibiting heightened susceptibility to SARS-CoV-2 without inducing brain damage, thus faithfully representing human COVID-19 pathology. Following infection with the mouse-adapted SARS-CoV-2 strain (MAp30), middle-aged hACE2-E1/2/8 mice exhibit more severe symptoms compared to their younger counterparts, with male mice showing more pronounced symptoms than females. Notably, middle-aged male mice develop long COVID-like symptoms, including persistent weight loss and sustained lung pathology post-acute infection recovery. Furthermore, our model replicates intricate immune responses observed in human COVID-19, such as heightened type I interferon levels and enhanced inflammation associated with neutrophils and eosinophils in plasma. Additionally, mice subjected to eosinophil or neutrophil depletion during early infection stages display milder disease symptoms. This model demonstrates substantial potential to facilitate in-depth exploration of COVID-19 and influence implications for therapeutic and preventative strategies.
Peptide-pulsed MHC class II allogeneic dendritic cell vaccine has superior efficacy providing allogeneic help in a murine cancer model.
Authors: Seishima Noriko , Becker William , Olkhanud Purevdorj , Maeng Hoyoung , Lopez-Lago Miguel , Wiseman Charles , Williams William , Berzofsky Jay 
Published: November 2024

Abstract
Allogeneic dendritic cell (DC) cancer vaccines present a promising alternative to autologous counterparts, offering an “off-the-shelf” solution for multiple patients and providing additional allogeneic help. We assessed the efficacy of a semi-allogeneic DC vaccine in comparison to a syngeneic one for tumor suppression. C57BL/6 mice were inoculated subcutaneously with human papillomavirus E6 and E7-expressing TC-1 cells. Syngeneic bone marrow DCs (BMDCs) were generated from C57BL/6 and semi-allogeneic BMDCs with a point mutation on either MHC class I or II were generated from B6.C-H2-Kbm1/By and B6(C)-H2-Ab1bm12/KhEg, respectively. The TC-1-bearing mice were injected with syngeneic or semi-allogeneic BMDCs pulsed with H-2Db-restricted E743-77 peptide. Compared with saline control, the MHC-I mutant BMDC vaccine reduced tumor growth no better than the syngeneic one. However, the MHC-II mutant BMDC vaccine was significantly better at delaying tumor growth. CD4+ or CD8+ T cell depletion showed that the syngeneic BMDC vaccine worked independently of CD4+ T cells, but the enhanced activity of the MHC-II mutant BMDC vaccine was dependent on CD4+ T cells at an early stage. Surprisingly, later depletion of CD4+ T cells improved vaccine efficacy, and this was confirmed to be due to Treg depletion. Thus, MHC class II allogeneic BMDCs proved more effective by inducing early allogeneic CD4+ T cell help, and this effect can by further enhanced by Treg depletion at a later stage.
PD1 blockade improves survival and CD8+ cytotoxic capacity, without increasing inflammation, during normal microbial experience in old mice
Authors: Dahlquist Korbyn , Huggins Matthew , Yousefzadeh Matthew , Soto-Palma Carolina , Cholensky Stephanie , Pierson Mark , Smith Declan , Hamilton Sara , Camell Christina 
Published: November 2024

Abstract
The elderly and those with chronic diseases face a 2 to 10-fold higher risk of hospitalization and mortality post-infection compared with healthy adults. Disease severity in the elderly is often attributed to the aging of the immune system, including an expansion of exhausted T cells that contribute to inflammation and have impaired activation and function. It is unclear how exhausted T cells from aged individuals respond to infection and the impact of acute restoration of T cell activation produces functional cells that contribute to excessive inflammation. In a mouse model of normal microbial exposure (NME), old specific pathogen-free (SPF) mice exhibit 100% mortality when exposed to microbes ordinarily found in pet store mice. We show that old mice exposed to NME show increased expression of inflammatory pathways and elevated frequencies of a heterogenous exhausted CD8+ T cell pool that express different levels of PD1, TOX, and CXCR5. Treatment with an anti-PD1 monoclonal blocking antibody in old mice during NME significantly improves survival without altering the acute inflammatory response despite the potential for adverse events following checkpoint blockade. Anti-PD1-mediated survival depends on CD8+ but not CD4+ T cells. CD8+ PD1+ T cells from old mice given anti-PD1 have increased GzmB production. These data suggest a novel approach for reducing infection vulnerability in the aged by targeting CD8+ T cell exhaustion through checkpoint blockade immunotherapy.
Towards optimizing cancer immunotherapeutic index: anti-CD3 Mono-Fab, a novel T cell-based therapeutic approach with less auto-Immunogenic profile.
Authors: Okpasuo Onyekachi , Mahadevappa Chandrashekara , Huynh Hien , Cheatam Christa , Radhakrishnan Vijay , Zaghouani Habib , Franklin Craig , Schrum Adam , Gil Diana 
Published: November 2024

Abstract
Despite advancement of cancer therapeutic approaches with immune checkpoint inhibitors (ICIs) against several tumor types, its anti-tumor benefit is limited by the emergence of immune related adverse events (irAEs). irAEs with an autoimmune phenotype, show a wide spectrum of toxicities that affect almost all organ systems. Several studies suggest a positive correlation between irAEs and anti-tumor response, with higher levels of toxicities seen with combination therapies. Finding strategies that optimize efficacy with less toxicity has been in the frontline of immunotherapy. We have shown a novel therapeutic strategy: anti-CD3e Monovalent Fab fragment (Mono-Fab) that targets the TCR/CD3 complex, strengthening signaling response to moderately weak antigens. Anti-CD3e Mono-Fab as a single therapy and in synergy with combination ICIs is effective against metastatic melanomas. To determine its optimal therapeutic index, we utilized a cell mediated experimental autoimmune encephalomyelitis mouse model as a platform to gauge potential to cause irAEs in the clinic. In this model, anti-CTLA-4 mAb administration to mice increases severity of their EAE symptoms over those observed in mice treated with appropriate IgG control, while anti-CD3e mono-Fab treatment does not. Thus, Anti-CD3e Mono-Fab contrast as an efficient potentially non-toxic cancer immunotherapy when compared with anti-CTLA-4 antibodies that are known to cause irAEs in patients.
Recombinant Fasciola hepatica fatty acid binding protein (Fh15) ameliorates dextran sulfate sodium (DSS)-induced ulcerative colitis in mice.
Authors: Figueroa-Gispert María Del Mar , Ramos-Lugo Claudia , Malavé-Ocasio Carlimar , Espino Ana 
Published: November 2024

Abstract
Ulcerative colitis (UC) is the most common inflammatory bowel disease affecting 5 million individuals worldwide. The Fasciola hepatica-derived molecule termed Fh15 is a TLR4 antagonist that has shown to exhibit powerful anti-inflammatory properties in murine and non-human primate sepsis models. Since TLR4 is involved in UC-pathological mechanisms, we hypothesize that Fh15 will suppress the pathological presentations of UC in mice. To test this hypothesis, we induced UC in C57BL/6 mice by providing animals with 4% DSS drinking water for 7 days. Mice were allotted into three experimental groups and four control groups containing eight mice each (4 male and 4 female). The experimental groups received i.p. injections with 50ug, 100ug or 150ug Fh15 on days 1, 3 and 5 of DSS-drinking. Animals that only drank DSS-water or that only received injections with 50ug, 100ug, or 150ug Fh15 were used as positive and negative controls, respectively. Results demonstrated that Fh15-treated animals had significantly less disease activity index, lower histopathological score, and longer colon-length than DSS-control animals; being 50ug the optimal Fh15 dose to reduce the extension of colon inflammation in male mice. Fh15 also suppressed other inflammatory markers such as MPO, CHI3L1, S100, T cells and macrophages. Preliminary results indicate that Fh15 could be an excellent therapeutic alternative to ameliorate the pathological signs of UC in the mouse model.
Catalytic activity and autoprocessing of murine caspase-11 mediate noncanonical inflammasome assembly in response to cytosolic LPS
Authors: Wodzanowski Kimberly , Akuma Daniel , Schwartz Wertman Ronit , Exconde Patrick , Vazquez Marrero Victor , Odunze Chukwuma , Grubaugh Daniel , Shin Sunny , Taabazuing Cornelius , Brodsky Igor 
Published: November 2024

Abstract
Inflammatory caspases are cysteine protease zymogens whose activation following infection or cellular damage occurs within supramolecular organizing centers (SMOCs) known as inflammasomes. Inflammasomes recruit caspases to undergo proximity-induced autoprocessing into an enzymatically active form that cleaves downstream targets. Binding of bacterial LPS to its cytosolic sensor, caspase-11 (Casp11), promotes Casp11 aggregation within a high molecular weight complex known as the noncanonical inflammasome, where it is activated to cleave gasdermin D and induce pyroptosis. However, the cellular correlates of Casp11 oligomerization and whether Casp11 forms an LPS-induced SMOC within cells remain unknown. Expression of fluorescently labeled Casp11 in macrophages revealed that cytosolic LPS induced Casp11 speck formation. Unexpectedly, catalytic activity and autoprocessing were required for Casp11 to form LPS-induced specks in macrophages. Furthermore, both catalytic activity and autoprocessing were required for Casp11 speck formation in an ectopic expression system, and processing of Casp11 via ectopically expressed TEV protease was sufficient to induce Casp11 speck formation. These data reveal a previously undescribed role for Casp11 catalytic activity and autoprocessing in non-canonical inflammasome assembly, and shed new light on the molecular requirements for noncanonical inflammasome assembly in response to cytosolic LPS.