bluuebunny/crossref_metadata_2025_split huggingface dataset:
Potent and specific killing of SLE donor B Cells with AlloNK (AB-101), an allogeneic cord blood-derived NK cell therapy, in combination with anti-CD19 or anti-CD20 monoclonal antibodies
Authors: Rogers Paul , guerrettaz lisa , Teng Kun-Yu , Palomino La Torre Claudia , Conerty Amanda , Flynn Peter , Raymon Heather 
Published: November 2024

Abstract
B cells play a crucial role in the pathogenesis of SLE by producing autoantibodies that target self-antigens, leading to tissue damage and inflammation. Here we demonstrate proof of concept for the use of a NK cell therapy to enhance the depletion of SLE donor B cells in the presence of anti-CD19 or anti-CD20 antibodies by an antibody dependent cellular cytotoxicity (ADCC) mechanism. AlloNK™ is a non-engineered, allogeneic, off-the-shelf, cryopreserved NK cell product, currently being evaluated in a clinical trial in combination with rituximab in subjects with lupus nephritis. AlloNK, which has been optimized for ADCC, was tested in combination with anti-CD20 (rituximab, obinutuzumab) or anti-CD19 (tafasitamab) mAbs in a short-term ADCC assay to show specific killing of SLE donor B cells. PBMC were isolated from SLE donors and co-cultured with AlloNK at various effector to target (E:T) ratios and mAbs concentrations. After 4h, the percentage of caspase 3/7+ B and T cells was determined by flow cytometry. At a 1:1 E:T ratio, AlloNK mediated killing of B cells in the presence of obinutuzumab (range 79-95%), rituximab (range 19-62%), and tafasitamab (range 24-77%) in a dose-dependent manner. Killing was specific as no off-target apoptosis of T cells was observed. Taken together, these data suggest that AlloNK has the potential to be effective in combination with mAbs to induce deeper B cell depletion and improved efficacy, over the mAbs alone, in SLE.
Notch2 signaling instructs TLR-responsive programs in B cells
Authors: Londregan Jennifer , Gaudette Brian , Galli Francesca , Tursi Nicholas , Allman David 
Published: November 2024

Abstract
The efficiency with which B cells respond to activation signals varies depending on the subset. Whereas marginal zone (MZ) B cells exhibit rapid activation kinetics following stimulation, follicular (FO) B cells adopt delayed effector programs. Further, MZ B cells uniquely exhibit readiness for the plasma cell (PC) fate, differentiating in mere hours independent of mitosis or T cell help. Some of the many ways MZ B cells achieve poised responsiveness is through sustained Myc levels and mTORC1 activity. MZ B cells uniquely require constitutive signaling through the transmembrane receptor Notch2, but whether this signal underlies the functionally poised nature of these cells is not understood. To examine the consequences of forced Notch2 on B cells with no Notch requirement, we co-cultured stimulated FO B cells on Notch2 ligand-expressing OP9 stromal cells. Notch2 signals modified the response to TLR 3 and 4 stimuli by amplifying the time to first division and the propensity toward PC differentiation altogether. Intriguingly, TLR3 responsiveness in B cells appeared to be entirely Notch2-driven and dependent on the signaling adapter Bruton’s Tyrosine Kinase. Employing a published mathematical model for dividing lymphocytes, we found Notch2 signals significantly augmented the division rate and division destiny of activated B cells. Together, these observations demonstrate a novel role for Notch2 in instructing B cell response programs to TLR signals and their effector outcomes.
Mapping the role of ITK (interleukin 2 inducible T cell kinase) and TCR (T cell receptor) signal strength in determining the early fate of CD8 T cells using Nr4a3 “timed” fluorescent reporter
Authors: Hunkins Josh , Ono Masahiro , Perrenoud Loni , Schoenbach Mary , Berg Leslie 
Published: November 2024

Abstract
ITK plays a key role in the rate and magnitude of TCR-dependent transcription factor activity and early genetic programming that determines the extent of CD8 T cell effector and memory function. To study the extent of this role we crossed an OT1 transgenic ITK germline KO to the Nr4a3-Tocky timer model which produces a time dependent fluorescent reporter. Nr4a3 is highly expressed following TCR activation and thus is a good metric for TCR signal strength and duration. The Tocky protein fluoresces blue, at λ 466 and then, after ~7hrs, degrades to a red fluorescence at λ 583. OT-1 Rag1KO ITK (WT, +/-, and -/-) Nr4a3-Tocky bulk splenocytes were stimulated with the altered peptide ligand T4 Ova peptide at varying concentrations for 12/24 hours and sorted on Tocky+ Blue/Red and negative events. ITK -/- CD8s showed elevated CD69 and CD25 but were variable for Tocky expression. Both Tocky Negative and Tocky Blue sorted ITK -/- CD69+ CD8s expressed higher IRF8, cFos, cMyc, Bcl2, and TCF1/7 as well as lower IRF4 versus WT at 12 hours post-stimulation. Analyses of Tocky Blue/Red or persistently active CD8s showed WT having increased expression of TCF1 and similar expression of IRF8, cFos, Bcl2, Eomes and Egr2 compared to ITK -/-. Bcl2 and PDL1 are also highly expressed through all stages of TCR signaling in ITK -/- CD8s. These data indicate that differences in TCR signal strength with and without ITK can affect the time at which transcriptional networks arise after TCR stimulation.
Assessment of molecular mechanisms involved in T-cell receptor (TCR)-dependent metabolic switch into aerobic glycolysis
Authors: Baracho Gisele , Reznan Ali , Shi Sam , Aruna Ayer , Widmann Stephanie , Tyznik Aaron 
Published: November 2024

Abstract
Upon activation, T cells undergo metabolic adaptations to fulfill the energy and biosynthetic requirements for their rapid growth and proliferation. TCR engagement and CD28 co-stimulation trigger the initial metabolic shifts that enable T-cell activation including increased glucose uptake. PI3K signaling plays a pivotal role in regulating Glut-1, the primary glucose transporter in T cells. Using high-dimensional spectral flow cytometry, we found correlated expression between Glut-1 and granzyme B (GZMB) in subsets of in vitro-activated human T cells. Notably, the PI3K inhibitor Ly294002 abrogated TCR-dependent upregulation of Glut-1 abolishing Glut-1highGZMB+ T cells in the cultures. A comprehensive single-cell targeted mRNA profiling of over 56, 000 immune cells captured with the BD Rhapsody™ HT Xpress System further showed a TCR-dependent upregulation of GZMB and various metabolic regulators such as GAPDH, HMGB2 and TK1. The results provide additional details on the involvement of the glycolytic pathway in T cell activation and offer a deeper insight into the molecular mechanisms associated with TCR-dependent cell activation.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.BD and BD Rhapsody are trademark of Becton, Dickinson and Company or its affiliates. © 2023 BD. All rights reserved. NPM-2938
Characteristics of tumor-rejecting neoantigens identified in a physiologically relevant cutaneous squamous cell carcinoma model
Authors: Hastings Karen , Adams Anngela , Macy Anne , Borden Elizabeth , Herrmann Lauren , Brambley Chad , Ma Tao , Li Xing , Hughes Alysia , Roe Denise , Mangold Aaron , Buetow Kenneth , Wilson Melissa , Baker Brian 
Published: November 2024

Abstract
Challenges in identifying tumor-rejecting neoantigens limit the use of neoantigen-based immunotherapies to treat high mutational burden tumors, such as cutaneous squamous cell carcinoma (cSCC). Most mutations in human cSCC are unique to an individual tumor. Only a small portion of human tumors share neoantigens predicted to bind MHC, supporting the need for personalized strategies. We generated transplantable cell lines, from solar UV-induced cSCC tumors, which recapitulate the UV mutational signature and driver mutations found in human disease. Depletion of CD8 and CD4 T cells revealed a dominant role for CD8 T cells and a supporting role for CD4 T cells in constraining cSCC. Vaccination with irradiated cSCC cells completely protects from tumor challenge and is dependent on CD8 T cells. Neoantigens were prioritized based on predicted binding affinity, MHC presentation and expression. IFN-γ-secreting CD8 T cells recognizing two neoantigens were present in cSCC tumors and tumor-draining lymph nodes. Prophylactic vaccination with these neoantigens constrained cSCC growth. Structural modeling of tumor-rejecting neoantigens in cSCC and other cancer models identified an increased solvent accessible surface area of the mutated residue in tumor-rejecting neoantigens compared to non-immunogenic neoantigens, which is anticipated to improve TCR recognition. This work provides a conceptual basis for consideration of TCR recognition in the prioritization of tumor-rejecting neoantigens.
Characterizing immune-mediated neuronal microtubule destabilization and axonal transport defects in a mouse model of optic neuritis
Authors: Sommers Kelli , Dittel Bonnie 
Published: November 2024

Abstract
Multiple sclerosis (MS) is a central nervous system autoimmune disease that often presents with inflammation of the optic nerve, optic neuritis (ON). The mechanisms of neuronal damage in MS and ON are unknown. In vitro studies modeling MS neurodegeneration demonstrated that cytolytic lymphocytes drive microtubule destabilization in human induced pluripotent stem cell-derived spinal motor neurons. Microtubules are essential for axonal transport, a key function for neuron health. We hypothesize that immune-mediated neuronal microtubule destabilization drives development of ON through disruption of axonal transport. MOG35-55 T cell receptor transgenic mice were used as a model of ON. The non-invasive ophthalmologic technique optical coherence tomography (OCT) allows for confirmation of symptomatic ON. Using OCT, increasing severity of progressive ON was observed over 5 months. To examine transport efficiency, cholera toxin subunit B (CTB), an axonal transport tracer, was injected into the eyes of mice with ON. Optic nerves were collected and stained for β3-tubulin, a marker of microtubules, and imaged by immunofluorescence. Preliminary data found decreased β3-tubulin staining and markedly reduced CTB transport in the presence of immune cell infiltration in mice with ON. These data confirm that OCT can be used to monitor ON progression and provide evidence that immune-mediated neuronal microtubule destabilization causes axonal transport deficits that underly ON development.
Bacterial peptidoglycan drives synovial inflammation in osteoarthritis
Authors: Dickman Jacob , Holub Meaghan , Rouse Joseph , Danner Rebecca , Wahhab Amanda , Jutras Brandon , Edelstein Adam , Lochhead Robert 
Published: November 2024

Abstract
Peptidoglycan (PG) is a potent inflammatory mediator. We have previously demonstrated that PG is present in the synovial tissue of osteoarthritis (OA) patients but its role in OA inflammation remains unknown. We hypothesized that PG in synovial tissue drives inflammation in OA. Intraoperative synovial tissue and synovial fluid samples were obtained from 56 consecutive patients with OA, with no prior history of infection. PG in synovial tissue was detected by immunohistochemical (IHC) staining of serial sections of 10 mm2 of tissue with anti-PG antiserum. PG was found in 33/56 (59%) of the tissue samples using IHC, with a median of eight PG occurrences per 10 mm2 in PG-positive samples. Tissue inflammation and fibrosis were assessed by histopathology. Cellular localization was further characterized in three PG-positive tissue samples by immunofluorescent microscopy (IFM). PG was localized to cells demonstrating mononuclear and fibroblastic morphology primarily in and around areas of vascularization and inflammation. CD68+ macrophages and to a lesser degree CD90+ synovial sublining fibroblasts. In multiplex assay analysis of synovial fluid cytokine levels, IL-6 and PG abundance were positively correlated. Fibroblasts and macrophages stimulated with PG in vitro secreted high levels of innate cytokines, paricularly IL-6 when measured by supernatant cytokine quantification. Together these data support the hypothesis that PG in synovial tissue drives joint inflammation in OA.
Peripheral mucosal-associated invariant T (MAIT) cells are severely depleted in HIV-positive heavy alcohol users
Authors: Li Wei , Khalili Mandana , Huang Laurence , Sanyal Arun , Shah Vijay , Chalasani Naga , Gupta Samir , Yu Qigui 
Published: November 2024

Abstract
MAIT cells are innate-like lymphocytes enriched in mucosal tissues, liver, and peripheral blood, contributing to protection against pathogens. Alcohol abuse and HIV infection independently reduce circulating MAIT cells. To assess their combined impact, we studied peripheral blood MAIT cells in heavy drinkers without overt liver disease (ALC), people with HIV (PWH) on ART, PWH on ART with heavy drinking (HIV-ALC), and healthy donors (HD). MAIT cell frequencies were significantly reduced in ALC (median 0.56, IQR 0.22-1.44; p=0.001), HIV (0.43, 0.20-0.97; p&lt;0.001), and HIV-ALC (0.30, 0.14-0.74; p&lt;0.001) vs HD (1.52, 0.74-2.71), accompanied by increased apoptotic MAIT cells. MAIT cells in HIV and HIV-ALC expressed elevated levels of activation markers (CD38, HLA-DR), with HIV-ALC also having higher CD69 levels than HIV and HD. Activated MAIT cells in HIV-ALC positively correlated with disease severity (prolonged prothrombin time, total bilirubin) and trended inversely with CD4% and CD4/CD8 ratio. However, no correlate was observed with cytokines linked to MAIT cell activation/function (IFN-α, IL-7, IL-12, IL-15, IL-17, IL-18, IL-23, IFN-γ, TNF-α). They also did not correlate with levels of blood biomarkers of gut injury/microbial translocation (TFF3, REG3α, LBP), monocyte/macrophage activation (sCD14, sCD163), and systemic inflammation (CRP). Conclusion: Peripheral blood MAIT cells in HIV-ALC were severely depleted, yet highly activated, and associated with disease severity.
Mechanistic insights into immune amnesia and immunity induced by measles infection and vaccination via comprehensive antiviral antibodies profiling
Authors: Chan Kwan Pui , Liu Liting , Bartlett Maggie , Griffin Diane 
Published: November 2024

Abstract
Measles is a disease known to induce acute immune amnesia and immunosuppression with increased susceptibility to other infections, while paradoxically conferring patients with life-long immunity to measles. However, it is not clear if the live-attenuated measles vaccine (LAMV) induces immunosuppression comparable to the wild-type measles (WT MeV) infection, nor how mechanistically LAMV and WT MeV affect antibodies specific to different pathogens.
Therefore, we utilized VirScan, a phage-display immunoprecipitation and sequencing (PhIP-seq) technology for virome-wide detection of antibodies against viral epitopes, to compare longitudinal antibody diversity in plasma samples from WT MeV and LAMV-infected rhesus macaques (RMs) (n=10 and 6 respectively, with 5 longitudinal timepoints at D0, D14/15, D28, D84/89, D112/168/176).
Preliminary analysis showed that WT MeV led to more long-term depletion of pre-existing antibodies to various viral peptides, while mounting a MeV-specific antibody response with greater intensity and breadth than LAMV-infected RM. Furthermore, in our sex difference analysis, male RMs elicited a stronger MeV response than female, particularly to N protein; also a more polyclonal MeV humoral response
In summary, our work provided crucial mechanistic insights into immune amnesia and immunity induced by natural infection versus vaccination, highlighting the public health value of the measles vaccine in averting MeV-induced immune amnesia.
Perforin production by T cells and Natural Killer cells in CMV-reactivated allogeneic cell transplant patients is associated with early post-transplant death
Authors: Sheikh Daanish , Nesher Lior , Khawaja Fareed , Shields Christina , Grimbergen Andrea , Nahmod Karen , Carrero Longlax Santiago , Ladki Malik , Nishiguchi Tomoki , Chemaly Roy , Makedonas George , DiNardo Andrew 
Published: November 2024

Abstract
Introduction:
Cytomegalovirus (CMV) is a prevalent herpesvirus that poses a serious risk to immunocompromised patients, particularly hematopoietic stem cell transplant (HSCT) recipients. CMV reactivation increases the risk of mortality due to secondary infections. CMV infection has been shown to epigenetically induce immune exhaustion and decrease non-specific immunity.
Methods:
We reviewed flow cytometry data from CMV seropositive HSCT recipients to determine if CMV reactivation predicted mortality via modulation of lymphocyte function. Peripheral blood mononuclear cells (PBMCs) were collected from recipients 30 days following transplant and stimulated with CMV-specific peptide pools of pp65 and staphylococcal enterotoxin B (SEB) to assess CMV-specific and non-specific immune function.
Results:
Non-specific, SEB-induced, polyfunctionality of CD4 and CD8 T cells was diminished in deceased CMV seropositive HSCT recipients. In deceased individuals, T-bet and perforin expression were elevated, with decreased IFNγ, TNFα, or IL2 SEB-induced production.
Discussion:
Mortality in CMV reactivated HSCT recipients was associated with non-specific immune exhaustion phenotypes, but it remains unclear whether this was a result of a higher pathogenic burden, recurrent CMV reactivation, or chemotherapy and myeloablation.
Microglial Differential Transcriptional Profiles in the Late-Stage Chronic Conditions of alpha synucleinopathy
Authors: Raihan Md Obayed , Fazal Nadeem , McGregor Brett A , Hur Junguk , Porter James E 
Published: November 2024

Abstract
Microglial immune function is compromised under pathological perturbation and a dysregulated immune response has been linked to neurodegenerative disorders and their progressive nature, including synucleinopathies. Using a transgenic mouse model of synucleinopathy (mThy1-hα-Synuclein), we have previously reported significant differences in microglial transcriptomic profiles at early and late stages of disease. We are interested in changes to microglia gene expression and regulation which contribute to disease progression or symptoms. Mice were characterized via behavior testing for motor deficit prior to microglia isolation and RNA extracted for RNA-sequencing. A total of 1,591 genes were differentially expressed (DEGs) in transgenic mice compared to age-matched controls. The DEGs were assessed for enriched functional terms via the Gene Ontology (GO) database and were clustered based on gene overlap between terms identifying 89 total clusters. Terms related to cytokine signaling, immune system, and synaptic transmission were enriched, which may suggest a link between declining motor function and αSyn associated pathology. Overall, our present study detailed the profile of αSyn reactive microglial under late-stage chronic conditions. Future studies in microglia regulation in neurodegenerative diseases will be critical to determining their contribution to disease progression as well as for developing novel therapeutics aimed at targeting neuroinflammation.
Natural History study to characterize CC011Unc/J spontaneous Mouse Model of Colitis
Authors: Gonzalo Gil Elena , Bowden Mia , Kramer Joshua , Davis Katherine , Copeland Katherine , Gutierrez Alan , Gott Rachel , Kasper Rachel , Case Laure 
Published: November 2024

Abstract
Objective: Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn’s disease, is a complex disorder characterized as an autoimmune response to the host’s intestinal microflora. We evaluate the progression of the colitis using CC01UncJ mice, a recombinant inbred mouse with similar characteristics to human IBD (Rogala AR et al., Mamm Genome, 2014. 25:95-108).
Methods: CC011Unc/J mice (JAX stock# 018854) were monitored weekly. Mice were euthanized at 8, 25, 30, 35 or 40 weeks of age, and colons were fixed and stained with hematoxylin and eosin. Slides were analyzed for leukocyte density, glandular hyperplasia, goblet cell loss, vasculitis, crypt abscesses, mucinous lakes, ulceration, and extent of lesion on a scale from 0-4.
Results: CC011Unc/J mice showed mild to moderate inflammation characterized by multifocal lymphocytes and plasma cell aggregates. A statistically significant scoring increase was observed between 8 weeks and 30 weeks (p&lt;0.01), with a plateau reached at 35-40 weeks of age. Significant differences between males and females were not observed for any of the scored parameters.
Conclusion: In contrast to the publications, CC011Unc/J mice showed only a moderate inflammation, suggesting an important role of environmental conditions to the development of the disease. Additional studies will be performed at JAX preclinical services to fully characterize these mice to offer this model as a platform for performing pre-clinical testing.
The function of DcR3.Fc and its derivatives in macrophage differentiation and skeletal muscle repair
Authors: Weng Chao-Yuan , Sung Pei-Shan , Hsieh Shie-Liang 
Published: November 2024

Abstract
Macrophages play a crucial role in regulating inflammation and tissue repair, influencing whether the tissue will regenerate or undergo fibrosis and dysfunction. However, the specific factors that can induce tissue repair/resolving macrophages remain unclear. Decoy receptor 3 (DcR3) is a soluble receptor without a transmembrane domain that can neutralize the effects of the tumor necrosis factor superfamily members FasL, LIGHT, and TL1A. In our previous study, we discovered that recombinant DcR3.Fc not only possesses neutralizing capabilities but also promotes M2-like macrophage differentiation. To delve deeper into its mechanisms, we created GRC2.Fc, which lacks the ligand-binding domain, thereby excluding the involvement of DcR3 ligands. Here, we observed that DcR3.Fc and GRC2.Fc induces differentiation of human CD14+ monocyte-derived macrophages and mouse bone marrow-derived macrophages toward a tissue repair/resolving phenotype in vitro. Furthermore, in a cardiotoxin-induced muscle injury model, intramuscular injection of GRC2.Fc protein promotes skeletal muscle repair and induce the expression of M2 macrophage markers in vivo. Our findings indicate that GRC2.Fc can induce macrophages toward a tissue repair/resolving phenotype and promote skeletal muscle repair.
Molecular Pixelation - dissecting the spatial organization of 80 surface proteins in single immune cells
Authors: van Ooijen Hanna , Karlsson Max , van Hoef Vincent , Schiller AnnaLotta , Thiagarajan Divya , Leijonancker Louise , Schweitzer Maud , Sathe Prajakta , Branting Annika , Martinez Barrio Alvaro , Karlsson Filip , Fredriksson Simon 
Published: November 2024

Abstract
Immune cell function is dependent on the activity of membrane proteins. Most widely-used cell analysis methods, flow cytometry and scRNAseq, directly or indirectly assess membrane protein abundance, yet provide no information about their spatial distribution. The gold standard technology for spatial protein analysis, fluorescence microscopy, has revealed that the spatial organization of membrane proteins is in fact of importance for their function, as exemplified by the immune synapse and uropods in migrating cells. Limited multiplexing and throughput scalability of microscopy, reduces its potential for forefront spatial proteomics discoveries, highlighting the need for new innovative methods.
Here, we present Molecular Pixelation (MPX), an advanced DNA barcode-based method for high-multiplex spatial analysis of proteins in single cells. Combining an NGS-based readout with our dedicated software, Pixelator, MPX enables the reconstruction of 3D cell-surface maps of 80 proteins on over 1000 individual cells, simultaneously. We applied MPX to compare the surface proteome of single resting and in-vitro activated T cells, highlighting differences in protein abundance, as well as revealing dramatic spatial protein re-organization, both in terms of supramolecular clustering and protein-protein co-localization. These results demonstrate that MPX has the potential to pave the way for expanding our understanding of cell surface protein architecture and its role in T cell activation.