bluuebunny/crossref_metadata_2025_split huggingface dataset:
Glycolipid-loaded Nanoparticles Harness iNKT cells for Tumor Immunotherapy
Authors: Shute Travis J , Dudley Elizabeth , Naranjo Joshua , Griffith Wendell , Nash Kelly , Leadbetter Elizabeth 
Published: February 2023

Abstract
Invariant natural killer T cells (iNKT) have a well-documented role in anti-tumor immunity through their release of proinflammatory cytokines and cytotoxic compounds. As iNKT cells can have direct and indirect killing effects on tumor cells, we propose a novel strategy for activating iNKT cells, via a PLGA nanoparticle delivery platform, to promote anti-tumor immune responses. Poly-lactic-co-glycolic acid (PLGA) nanoparticles can be reproducibly loaded with an iNKT cell glycolipid agonist, alpha-galactosylceramide (alpha-GalCer), and a model tumor associated antigen, ovalbumin (OVA). These dual-loaded PLGA nanoparticles rapidly activate iNKT cells in vivo to produce IFNgamma. Furthermore, in an in vivo model of melanoma, using B16F10-OVA cells, both prophylactic and therapeutic administration of nanoparticles containing alpha-GalCer and OVA led to decreased tumor cell growth and increased survival. We also show our nanoparticle therapy has synergistic potential with two immune checkpoint blockade therapies, anti-PD-1 and anti-CTLA-4, currently used in the clinic for human cancer patients. This novel delivery system provides a platform with tremendous potential to harness iNKT cells for cancer immunotherapy purposes against many different cancer types.
The mRNA-LNP platform’s lipid nanoparticle component used in preclinical vaccine studies is highly inflammatory
Authors: Ndeupen Sonia , Qin Zhen , Jacobsen Sonya , Igyarto Botond Z 
Published: February 2023

Abstract
Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against coronavirus disease in 2019 (COVID-19). Clinical trials and ongoing vaccinations present with very high protection levels with varying degrees of side effects. However, the nature of the reported side effects remains poorly defined. Here we present evidence that LNPs used in many preclinical studies are highly inflammatory in mice. Intradermal injection of these LNPs led to rapid and robust inflammatory responses, characterized by massive neutrophil infiltration, activation of diverse inflammatory pathways, and production of various inflammatory cytokines and chemokines. The same dose of LNP delivered intranasally led to similar inflammatory responses in the lung and resulted in a high mortality rate.
In summary, here we show that the LNPs used for many preclinical studies are highly inflammatory. Thus, their potent adjuvant activity and reported superiority comparing to other adjuvants in supporting the induction of adaptive immune responses could stem from their inflammatory nature. Furthermore, the preclinical LNPs are similar to the ones used for human vaccines, which could also explain the observed side effects in humans using this platform.
Co-immunization with adenosine deaminase rescues age-associated impairment of SARS-COV-2 synDNA vaccine-induced responses
Authors: Gary Ebony Nicole , Connors Jennifer , Tursi Nicholas J. , Warner Bryce , Bell Matthew , Cuismano Gina , Taramangalam Bhavani , Zhang Shiyu , Canziani Gabriela , Chaiken Irwin C. , Wootton Sarah , Weiner David , Kutzler Michele A. , Haddad Elias , Kobasa Darwyn 
Published: February 2023

Abstract
SARS-CoV-2 is responsible for a global pandemic claiming over 2 million lives and infecting over 100 million people. Elderly patients display increased COVID-19 morbidity and mortality. Vaccine candidates have been studied and deployed in the clinic. However, age-associated immune deficits are known to cause sub-optimal responses, and the longevity of vaccine-induced responses is under investigation. Germinal center follicular helper T cells (TFH) promote affinity maturation of B cell receptors and memory B cell differentiation and are defined by expression of adenosine deaminase-1 (ADA-1). We investigated the adjuvant properties of plasmid-encoded adenosine deaminase (pADA) in the context of a SARS-CoV-2 spike synDNA antigens. Young and aged mice were immunized with plasmid-encoded spike (pS) alone or co-immunized with pS and pADA and cellular and humoral responses were evaluated. When immunized with pS alone, aged mice had decreased spike-binding IgG and neutralization titers. However, young and aged animals co-immunized with pADA had similar humoral responses. We observed similar trends in serum antibody affinity as measured by SPR. pADA co-immunization also rescued age-associated decreases in spike-specific IFNy secretion in the spleens and lungs of co-immunized aged mice as measured by ELISpot. Finally, preliminary data analysis indicates that co-immunization with pADA significantly impacts viral load in a model of SARS-CoV-2 infection. These data suggest that pADA enhances antigen-specific cellular and humoral immunity in aged mice and supports further study of this molecule as an immunoadjuvant for vaccines targeting elderly populations.
An adjuvant strategy enabled by modulation of the physical properties of fungal mannans elicits pan-coronavirus reactive anti-SARS-CoV-2 Spike antibodies
Authors: Borriello Francesco , Spreafico Roberto , Poli Valentina , Shrock Ellen , Chou Janet , Barrett Nora A. , Lacanfora Lucrezia , Franco Marcella E. , Marongiu Laura , Iwakura Yoichiro , Pucci Ferdinando , Kruppa Michael D. , Ma Zuchao , Lowman Douglas W. , Ensley Harry E. , Nanishi Etsuro , Saito Yoshine , O’Meara Timothy R. , Seo Hyuk-Soo , McGrath Marisa E. , Logue James , Haupt Robert E. , Dhe-Paganon Sirano , Dowling David J. , Frieman Matthew , Elledge Stephen J. , Levy Ofer , Irvine Darrell J. , Williams David L. , Zanoni Ivan 
Published: February 2023

Abstract
Activation of the innate immune system via pattern recognition receptors (PRRs) is key to generate long-lasting adaptive immunity. While it is known that PRRs detect unique chemical patterns associated with invading microorganism, if and how the physical properties of PRR ligands influence development of the immune response is largely overlooked. Through the study of fungal mannans we present data that put the physical form of PRR ligands at the center of the process that determines the outcome of the immune response. Soluble mannans are immunosilent in the periphery but elicit a potent pro-inflammatory response in the draining lymph node (dLN). By modulating the physical form of mannans, we developed a formulation that targets both periphery and dLN. When combined with SARS-CoV-2 Spike, this formulation elicits neutralizing anti-SARS-CoV-2 antibodies that cross-react with pathogenic coronaviruses. Thus, the physical properties of fungal ligands can be harnessed for rational adjuvant design and vaccine development.
Optimizing immunoassay-based selection of donors for COVID-19 convalescent plasma
Authors: Van Ert Hanora A , Rogers Kai , Bohan Dana , Han Chanze , Dempewolf Spencer , Sevcik Kristina , Ruggio Natalie , Rojas-Chavez Anthony , Boyt Devlin , Pentella Michael , Jackson Brooks , Merrill Anna , Knudson Michael , Maury Wendy , Haim Hillel 
Published: February 2023

Abstract
COVID-19 convalescent plasma (CCP) is currently administered to hospitalized COVID-19 patients under an EAU by the FDA. CCP units are selected by commercially available ELISA quantification of anti-SARS-CoV-2 (SARS2) antibodies under the rationale that higher levels of anti-SARS2 antibodies correlate with higher neutralizing capabilities and thus greater efficacy. The validity of this selection process has yet to be investigated. We analyzed serum samples from 60 convalesced donors with PCR-confirmed COVID-19. Samples were analyzed using commercial SARS2 ELISAs (DiaSorin, Euroimmun, Ortho, and Roche) and a cell-based enzyme-linked immunosorbent assay (cbELISA) that detects antibodies against the native conformation of the SARS2 spike glycoprotein (GP). Sera were also tested for neutralizing activity against wild-type SARS2 and a pseudovirus expressing the SARS2 GP. Immunoassay and neutralization values were compared to determine the precision of each immunoassay to predict differing levels of neutralizing activity, and ability of each immunoassay to identify high neutralizing activity. Results of immunoassays that apply SARS2 GP as the antigen (DiaSorin, Euroimmun, Ortho, and cbELISA) correlated well with neutralization titers measured by wild-type SARS2 or SARS2 GP-pseudotyped virus as compared with assays using other SARS2 antigens (Roche; N protein). Differences in precision of these assays were evident when samples within the top quintile or decile of immunoassay values were analyzed, with DiaSorin and Ortho assays showing superior performance in predicting neutralizing capability. We hope this approach will assist in the selection of highly neutralizing CCP units that confer greater clinical efficacy.
EGR2 and EGR3 function in a cell-intrinsic and partially redundant manner to suppress the accumulation of B1a cells and CD21low CD23low age-associated B cells
Authors: Masle-Farquhar Etienne , Peters Timothy , Oakes Christopher , Reed Joanne , Goodnow Christopher C 
Published: February 2023

Abstract
Egr2 and Egr3 genes encode early growth response (EGR) transcription factors, act as immediate early genes following B cell receptor (BCR) signalling and are highly up-regulated in anergic mouse B cells. Anergy preserves microbial epitope responsiveness from a finite pool of pre-immune B cells, but is also reversible – creating a risk of autoimmune disease. Pathological proliferation of self-reactive B cells can also cause chronic lymphocytic leukemia (CLL), 3.8% of which harbour somatic missense EGR2 mutations resulting in loss- or change-of-function and correlated with poor prognosis. We analysed B cells in mice lacking one or both alleles of Egr2 and/or Egr3 and show that Egr2 and Egr3 deletion cause the cell-intrinsic accumulation in spleen, blood and bone marrow of populations enriched for self-reactive BCRs: B1a and CD21low CD23low age-associated B cells. Global single-cell RNA profiling of these expanded populations in vivo demonstrated their differential expression of genes involved in their survival and maintenance. We used chromatin immunoprecipitation sequencing (ChIP-Seq) to show that several of these genes are direct EGR2 transcriptional targets in human CLL. This is the first report on the roles of Egr2/3 in B cells and we demonstrate herein that Egr2 and Egr3 transcription factors are crucial in suppressing the accumulation of B1a and age-associated or atypical memory B cells.
The role of berberine, a clinically relevant plant-derived alkaloid, in T-cell mediated immunosuppression
Authors: Vita Alexandra Adorno , Pullen Nicholas A. 
Published: February 2023

Abstract
Preliminary research revealed a novel function of berberine (BBR), a clinically relevant plant-derived alkaloid, as a suppressor of follicular T helper (Tfh) cell proliferation in secondary lymphoid organs of BBR-treated mice that underwent immunization for arthritis induction in a collagen induced arthritis (CIA) mouse model, a common murine model for rheumatoid arthritis. As the interaction between Tfh cells and B cells is crucial for thymus-dependent humoral responses, this raises concern that the prolonged use of berberine could suppress the generation of such responses. In our current research we explored the effects of BBR in vitro on the differentiation of Tfh cells from naïve CD4+T cells (&gt;95% pure) by examining the expression of the key Tfh cell surface molecules CXCR5, ICOS, and PD1 following BBR treatment. Such molecules are crucial for Tfh cell effector function. The treatment of BBR at 0.25 mg/mL, 0.50 μg/mL, and 1 mg/mL significantly reduced the expression of both CXCR5 (p &lt;0.01) and ICOS (p &lt; 0.01), but not PD1. It is important to note that in preliminary dose-response experiments, 0.50 mg/mL was the highest dose of BBR that did not significantly lead to cell death, indicating that the decreased expression of CXCR5 and ICOS at 0.25 mg/mL and 0.50 mg/mL are likely not due to cytotoxic effects. In the future, we plan to elucidate the mechanism by which BBR inhibits the expression of CXCR5 and ICOS by examining key cell signaling pathways that lead to Tfh activation and differentiation, and ultimately to CXCR5 and ICOS expression, following BBR treatment.
Expansion of T follicular regulatory cells by OX40L-JAG1 co-signaling ameliorates experimental lupus nephritis
Authors: Kumar Prabhakaran , Setty Suman , Epstein Alan L , Chiba Shigeru , Prabhakar Bellur S 
Published: February 2023

Abstract
Antinuclear autoantibodies (Nabs) promote tissue inflammation involving multiple organs including the kidneys in Systemic Lupus Erythmatosus (SLE). T-follicular helper (TFH) cell-dependent B-cell response plays an essential role in lupus nephritis (LN). TFH and B-cells receive mutual help in the germinal centers (GCs) to enable high-affinity autoantibody production. T follicular regulatory cells (TFR) cells, a specialized subset of CD4+Foxp3+Tregs, suppress GC formation and TFH-dependent B-cell response. Previously, we have shown that the expansion of functional Tregs using soluble OX40L and JAG1 proteins suppressed experimental autoimmune diabetes and thyroiditis. However, unlike other autoimmune diseases, the role of Tregs in lupus remains unresolved. Here, we show that NZBWF1/j mice with severe proteinuria and higher anti-dsDNA IgG levels had increased GC-TFH, GC-B cells, and GC-TFH/GC-TFR ratio in their renal lymph nodes compared to mice with no proteinuria. While total Tregs were significantly higher in mice with proteinuria, GC-TFRs were significantly reduced. Intriguingly, OX40L-JAG1 treatment significantly increased Tregs, GC-TFRs, and reduced GC-TFH/GC-TFR ratio. Periodic OX40L-JAG1 treatment resulted in a significant reduction in GC-B-cells, anti-dsDNA IgG level, proteinuria and kidney pathology, and increased survival. Mechanistic studies showed increased PD1 expression in Tconv cells, Tregs, TFH, and TFR cells, and PD-L1 expression in GC-B-cells suggesting a key role for PD1 signaling in the observed protective effect. Our results show that OX40L-JAG1 treatment expanded TFRs which inhibited GC-B cell activation and NAb production, proteinuria and LN.
T-cell receptor repertoire of mice with organ-specific autoimmunity resulting from a partial defect in T cell negative selection and dendritic cell enhancement
Authors: Yin Mianmian , Smith Jennifer A , Chou Marissa , Chan Jackie , Hu Yongmei , Lowell Clifford A , Gould Douglas B , Anderson Mark S , DeFranco Anthony L 
Published: February 2023

Abstract
Previously, we reported a new genetic model of spontaneous autoimmune disease in which mice with a hypomorphic mutation of Aire (AireGW/+) combined with deletion of Lyn in dendritic cells develop autoimmune uveitis with a 50% penetrance. Here we demonstrate that a key autoantigen in this model is interphotoreceptor retinoid-binding protein (IRBP) as AireGW/+ Lyn−/− IRBP−/− mice failed to develop uveitis. The expansion of CD4+ T cells recognizing amino acids 271–290 (“P2”) of IRBP correlated strongly with autoimmune disease in mice with intact IRBP. To study the role of these T cells, we did single-cell RNA-seq with TCR sequencing for P2-specific CD4+ T cells isolated from eye-draining lymph nodes (LN) of AireGW/+ Lyn−/− mice with and without uveitis. Both mice showed an expansion of these T cells in eye-draining LNs, but it was greater in mice with disease. A remarkable clonal diversity of TCRs was seen, with some sharing of TCR Vα or Vβ sequences. Expansions of particular T cell clonotypes were more evident in LN of mice with disease, and even greater in the retinas of mice with disease. We reconstituted several P2–specific TCR clonotypes in a T cell hybridoma line and verified their specificity. One of the TCR clonotypes was expressed in developing thymocytes by retroviral transduction of bone marrow stem cells from Rag2−/− mice and transplanted into WT or AireGW/+ recipient mice. Remarkably, the small residual expression of IRBP in the thymus of AireGW/+ mice was sufficient to induce substantial negative selection of thymocytes with this TCR clonotype. Thus, deficiency of negative selection of P2 tetramer–specific TCR promotes the development of uveitis in AireGW/+ Lyn−/− mice, even if negative selection is only partly compromised.
ICOS is critical for T follicular regulatory cell differentiation
Authors: Panneton Vincent , Mindt Barbara , Bouklouch Yasser , Chang Jinsam , Witalis Mariko , Li Joanna , Stancescu Albert , Bouchard Antoine , Bradley John , Randall Troy D , Fritz Jörg H , Suh Woong-Kyung 
Published: February 2023

Abstract
The Inducible Costimulator (ICOS) is a T cell costimulatory receptor critical for humoral immunity. ICOS-deficient patients suffer from recurrent infections due to lack of protective antibodies. However, some patients also display signs of antibody-mediated autoimmunity. These findings may reflect a dual role of ICOS: facilitating the differentiation and function of T follicular helper (Tfh) and T follicular regulatory (Tfr) cells. While Tfh cells are known to provide help to B cells to produce high affinity antibodies, the main role of Tfr cells seems to be preventing autoantibody generation. Using Foxp3-cre-mediated ICOS knockout (ICOS FC) mice, we show that T regulatory (Treg)-specific ICOS deletion drastically reduces the number of Tfr cells without altering Treg cell numbers. Single cell RNA sequencing further revealed shifts in transitory Tfr precursor populations in immunized ICOS FC mice. Importantly, we observed a lowered ratio of antigen-specific germinal center B (GCB) cells and increased anti-nuclear antibodies in ICOS FC mice, suggesting a rise in autoreactive GCB cells. We also noted variations in isotype composition of total and virus-specific antibodies in infected ICOS FC mice. Mechanistically, our data suggests that ICOS could promote the Treg-to-Tfr transition by regulating CXCR5 expression. Thus, our study demonstrates that ICOS is critical for Tfr cell generation and supports the role of Tfr cells in preventing generation of autoantibodies during germinal center reactions.
AIM2 in regulatory T cells restrains autoimmune diseases
Authors: Chou Wei-Chun , Guo Zengli , Wan Yisong , Ting Jenny P-Y 
Published: February 2023

Abstract
The inflammasome comprised of an innate immune receptor/sensor, pro-caspase-1, and a common adaptor molecule, ASC (apoptotic speck-containing protein with a CARD) initiates innate defense and inflammatory response by activating caspase-1 and pyroptotic cell death in myeloid cells. Consistent with their pro-inflammatory function, caspase-1, ASC and NLRP3 were known to exacerbate autoimmunity during experimental autoimmune encephalomyelitis (EAE) by enhancing IL-1b and IL-18 secretion in myeloid cells. Here we reveal an unexpected function of a DNA-binding inflammasome receptor, AIM2 (Absent in Melanoma 2), in T regulatory cells (Tregs) to restrain two models of autoimmune disease, including experimental autoimmune encephalomyelitis and T cell-mediated colitis, by studying whole-body and Treg-specific Aim2-deficient mice. AIM2 is highly expressed by human and mouse Tregs, with its expression induced by TGF-b and its promoter occupied by transcription factors associated with Tregs, including Runx1, Ets1, Bcl11b and CREB. Moreover, we demonstrate that AIM2 promotes the stability of Tregs during inflammation in lineage tracing mice. RNA-seq, biochemical and metabolic analyses revealed that AIM2 attenuates Akt-phosphorylation, mTOR, Myc and glycolysis, but promotes lipid oxidative phosphorylation in Tregs. Mechanistically, AIM2 interacts with the RACK1/PP2A-phosphatase complex to restrain Akt-phosphorylation. While AIM2 is generally accepted as an inflammasome effector in myeloid cells, this report unveils a T cell-intrinsic role of AIM2 in restraining autoimmunity by diminishing Akt-mTOR signaling and altering immune-metabolism to enhance Treg stability.
Helios expression in CD4+Foxp3− T cells
Authors: Tison Kole , Penna Vinay , Skadow Mathias , Oguz Cihan , Lack Justin , Thornton Angela M , Shevach Ethan M 
Published: February 2023

Abstract
The transcription factor Helios is expressed in the majority of Foxp3+ Treg cells, where it is necessary for the maintenance of self-tolerance. Mice with a Foxp3-specific deficiency in Helios develop systemic immune activation with increased T follicular helper (Tfh) and T helper 1 (Th1) effector responses. Notably, mice with a CD4-specific deletion of Helios do not exhibit autoimmunity, pointing to an additional role for Helios in CD4+ T cells. In fact, Helios is expressed in a minor population of conventional CD4+ T cells and its expression is upregulated upon activation of CD4+ T cells, yet the role of Helios in these cells remains unknown. We generated TCR Tg mice with a CD4-specific Helios deletion and have shown that Helios deficient naive CD4+ T cells are fully able to differentiate into Th1, Th2 and Tfh effector cells in vivo. Helios deficient CD4+T cells show no defect in expansion or the production of cytokines. However, Helios deficiency increased the propensity of naive T cells to differentiate into antigen-specific pTregs under inflammatory as well as tolerizing conditions and attenuated the induction of a memory response. To further characterize CD4+Foxp3−Helios+ cells, Next-generation sequencing (NGS) analysis was performed. This analysis has yielded evidence for a heterogenous population that is comprised of Tfh, Tmem, iNKT and a population of cells that have Treg-like features. In vitro activation assays and qPCR have validated differentially expressed cytokines between Helios+ and Helios− CD4+ T cell populations, in agreement with the NGS data. Together, these findings suggest that Helios may play a role in stabilizing effector function.
Kidney macrophages in human lupus nephritis and mouse models express conserved inflammatory and remodeling gene programs and localize to the same anatomic sub-compartments.
Authors: Hoover Paul , Peters Michael , Lieb David , Jones Tony , Mishra Rakesh , Hacohen Nir , Davidson Anne 
Published: February 2023

Abstract
Tissue macrophages promote kidney homeostasis through tightly regulated initiation, maintenance, and resolution of tissue repair. In lupus nephritis (LN), kidney macrophages may drive damage in glomeruli and other kidney sub-compartments leading to proteinuria and fibrosis. Recent single-cell RNA sequencing work uncovered 4 novel kidney macrophage subsets in LN patient biopsies: M2-like “reparative”, “inflammatory”, “phagocytic”, and “resident” subsets. But how macrophage gene programs translate into cellular functions driving proximal tissue damage has been difficult to study because we do not fully understand how mouse models recapitulate human LN. Here, we used single cell RNA sequencing to identify kidney macrophage subsets conserved in human and mouse models of LN. We compared the transcriptomes of macrophages collected from human LN patients at peak clinical disease to those from 4 mouse strains at early and peak clinical disease. We discovered that human “reparative”, “inflammatory”, “phagocytic” subsets were conserved across species. In particular, human “reparative” and mouse resident macrophages shared inflammatory and remodeling programs, expressed interferon stimulated genes, and localized to the same kidney sub-compartments. In nephritic mice, this subset converted to a mixed M1-inflammatory/M2-repair phenotype also observed in humans, thus identifying potential conserved programs driving tissue damage in mice and humans. These findings strongly support shared roles for conserved human and mouse macrophages. Critically, they open a new path utilizing mice to study how conserved programs translate into cellular functions driving proximal tissue damage.