bluuebunny/crossref_metadata_2025_split huggingface dataset:
IFN-lambda orchestrates an IL-10 immunoregulatory program in dendritic cells critical for T cell-mediated protection against influenza virus infection
Authors: Hemann Emily A , Green Richard 
Published: February 2023

Abstract
Influenza virus (IAV) infection remains a major global health burden that requires development therapeutic and vaccination strategies to limit infection and drive robust and durable protective immune responses. Type III interferon (IFN), IFN-lambda, restricts IAV infection in vivo compared to type I IFN. However, the contribution of IFN-lambda to generating adaptive immune responses against IAV remains unknown. Utilizing a murine model of IAV infection in mice lacking the IFN-lambda receptor (Ifnlr1−/−), we determined that IFN-lambda signaling is required for full induction of effector and memory CD8 T cell responses Ifnlr1−/− are significantly more susceptible to mortality following heterosubtypic IAV re-infection. This defect in the CD8 T cell response was not intrinsic to the T cells, but rather due to aberrant dendritic cell (DC) function in the absence of IFN-lambda signaling. Transcriptional analysis of conventional DCs (cDC) sorted from draining lymph nodes of IAV-infected mice revealed specific modulation of an immunoregulatory program in cDC from Ifnlr1−/−, but not WT mice. Indeed, we found IL-10 to be secreted only by Ifnlr1−/−, and not WT, BMDC following IAV infection. Addition of IL-10 to WT BMDC only partially recapitulated defects in DC maturation observed in Ifnlr1−/−, suggesting more additional pathways of regulation of DC by IFN-lambda. Our analysis further revealed shared and unique Ifnar1- and Ifnlr1-dependent transcriptional responses in DCs, highlighting the distinct role IFN-lambda plays in regulating DC function during IAV infection. Overall, these studies reveal a unique role for IFN-lambda programming of DCs to optimally activate protective CD8 T cell responses during influenza virus infection.
Anti-insulin B cell receptors display skewed immunoglobulin gene usage and limited mutation in pre-symptomatic type 1 diabetes donors
Authors: Scaglione Steven R , Joosse Bryan A , Westlake Camille , Kim Eunice , Santhin Austin B , Forchetti Mason V , Cisneros Alberto , Moore Daniel J , Smith Scott A , Bonami Rachel H 
Published: February 2023

Abstract
Antibodies targeting insulin and other islet antigens predict type 1 diabetes (T1D) onset. Insulin autoantibody (IAA)-positive individuals progress to diabetes more rapidly, yet little is known about how IAA-precursor anti-insulin B cells (AIBCs) recognize insulin. Complementarity-determining regions (CDRs) of the B cell receptor (BCR) are structurally diverse loops that constitute the antigen-binding site, and amino acid variability in BCRs increases the breadth of repertoire antigen-specificities. To investigate anti-insulin BCRs during the earliest detectable stages of T1D, we enrolled eight insulin therapy-naïve Type 1 Diabetes TrialNet Pathway to Prevention participants who were positive for ≥2 islet autoantibodies and thus at high risk for diabetes. We stimulated donor peripheral blood mononuclear cells to drive BCR secretion as antibody, screened for AIBC-containing wells by ELISA, and immortalized these cells as hybridomas. We validated 25 monoclonal anti-insulin hybridoma lines and sequenced 16 heavy chain (IgH) and 14 light chain (IgL) immunoglobulin genes. 25% of VH and 50% of VL sequences contained at least one mutation within a CDR compared to germline, with 13% of VH and 29% of VL sequences having ≥6 mutations within the V-region. We surveyed previously and currently reported AIBC VH (n=25 BCRs), which revealed skewed VH and JH gene use compared to the total polyclonal IgH repertoire (n= 8790 BCRs, p &lt; 0.001 and p &lt; 0.05, respectively, chi-squared test). These data expose unique features of insulin recognition among AIBCs isolated from pre-symptomatic T1D donors, including V and J gene usage bias and potential for germline IgH insulin recognition, providing genetic and molecular insight into AIBC origins.
Intra-islet CD8+ T cells are restrained by an exhaustion program that can be reversed by LAG3 deletion
Authors: Grebinoski Stephanie J , Zhang Qianxia , Cillo Anthony R , Das Jishnu , Workman Creg , Vignali Dario 
Published: February 2023

Abstract
CD8+ T cell exhaustion or dysfunction is a differentiation state in which T cells become unresponsive to stimulation and lose effector function1. Impaired chronic viral and tumor clearance has been attributed to CD8+ T cell exhaustion and can be partially reversed upon blockade of inhibitory receptors (IR) such as programmed cell death protein 1 (PD1). Despite detailed analysis of CD8+T cell in tumors and chronic viral infection, the role of the exhaustion program and the transcriptional networks that control CD8+T cell function and fate in autoimmunity are still unknown. Here we show that intra-islet CD8+T cells phenotypically and transcriptionally possess features of exhausted T cells, yet maintain key differences, such as an expanded repertoire of effector mechanisms, that contribute to their persistent pathogenicity in autoimmune diabetes. This dual identity, which we refer to as a ‘divergent’ exhaustion program, can be perturbed substantively by CD8+ T cell-restricted deletion of the IR Lymphocyte Activating Gene 3 (LAG3), causing enhanced effector-like function and pathogenicity. Understanding this divergent exhaustion program and its key mediators will guide future immunotherapeutic interventions by characterizing ways to promote exhaustion in autoimmunity and reversing exhaustion programs in chronic viral infections and tumors.
Impact of B cell-depletion on plasma cells in a pre-clinical model of meningeal inflammation and subpial demyelination
Authors: Wang Angela , Ward Lesley , Ramaglia Valeria , Urich Eduard , Gommerman Jennifer L 
Published: February 2023

Abstract
Anti-CD20 B cell depletion therapy has provided evidence for the pathogenic roles of B cells in multiple sclerosis (MS). Terminally differentiated plasma cells (PCs) are spared by anti-CD20, and our lab has previously shown that during experimental autoimmune encephalomyelitis (EAE), IgA+ PCs mediate protection against neuroinflammation. One hypothesis is that in addition to depleting pro-inflammatory B cells, anti-CD20 also removes the major consumers of BAFF (i.e. B cells) thus promoting a favourable niche for regulatory PCs and may represent an additional mechanism by which anti-CD20 therapy confers protection.
Adoptive transfer EAE of PLP-primed Th17 cells into SJL/J mice leads to ascending paralysis and meningeal inflammation which parallels brain pathology seen in human MS. Mice were prophylactically treated with anti-CD20 (10mg/kg) or isotype control antibodies which we re-administered throughout the course of the experiment and monitored daily for clinical symptoms. We find that anti-CD20 treatment effectively depleted B cells (but not T cells) in the periphery; within the central nervous system, meningeal immune clusters formed in both anti-CD20 and isotype control treated mice, with the former being devoid of B220+ B cells. Compared to control treated mice, anti-CD20 treated mice showed limited subpial demyelination and pathology adjacent to B cell-less meningeal immune clusters, and is accompanied by a reduction in clinical score. In the serum, anti-CD20 treatment elevated levels of BAFF and IgA, which inversely correlated with disease severity. We thus propose that anti-CD20 exerts its protective effects by providing a favourable niche for suppressive IgA+ PCs.
Evaluation of differential autoantibody profiles in gut and brain tissues in children with Autism Spectrum Disorder (ASD).
Authors: Celis Nora Alexandra Ramirez , Van de Water Judy 
Published: February 2023

Abstract
Autoimmunity is a comorbidity often associated with autism spectrum disorder (ASD), and a better understanding of immune dysbiosis and autism is crucial to identify differential ASD phenotypes and develop better biomarkers. We aimed to study autoantibody (aAb) reactivity to gut and brain tissues to find aAb profiles in children with autism that could serve to identify ASD subpopulations with biomarker potential.

Methods:
We used plasma from typically developing children (TD; n=149) and children diagnosed with ASD (n=208). To assess aAb reactivity, we probed plasma brain and gut tissues by Western blot.

Results:
While we found reactivity against brain (28% ASD and 32% TD), and gut (15% ASD and 11% TD) in both groups, we observed reactivity to antigens in defined molecular weight ranges in the gut tissue recognized by aAbs from the ASD subjects and not the TD samples. Further, some individuals had aAb reactivity to both brain and gut (8.6% ASD and 8% TD). These data are consistent with our previous studies against cerebellum tissue, where we reported aAb reactivity in both groups (ASD and TD), but some autoantibodies were differentially reactive in the ASD population.

Conclusion:
We identified aAbs reactive to brain and gut antigens in both TD and ASD children, with differential recognition of plasma from children with ASD to proteins in the gut extract. We are currently working on identification of the differentially recognized proteins in the gut tissue. Further, we can link our findings to the clinical data of these children, including reported GI issues. Such aAbs have the potential to serve as predictors of a subgroup of ASD individuals or facilitate a better understanding of the different sub-phenotypes of ASD based on differential aAb profiles.
Interleukin-38 promotes skin tumorigenesis via regulation of epidermal cell hyperplasia and pro-tumorigenic microenvironment in an IL-1Rrp2-dependent manner.
Authors: Zhou Hong , Zhao Qi-xiang , Yu Jia-dong , Yue Cheng-cheng , Hao Yan , Zheng Hua-ping , Hu Jing , Wang Zhen , Li Jiong 
Published: February 2023

Abstract
As the newest member of the IL-1 family, interleukin-38 (IL-38) has strong associations with chronic inflammatory diseases. However, its roles in tumorigenesis and the underlying mechanism remain poorly understood. Here, we demonstrated that IL-38, which was highly expressed in skin, downregulated in human cutaneous squamous cell carcinomas (CSCC) and 7,12-dimethylbenzanthracene (DMBA)/12-O-tetradecanoyl phorbol-13-acetate (TPA) induced mouse skin tumorigenesis. IL-38 keratinocyte-specific knockout mice (IL-38 cKO) dramatically promoted skin tumor formation and their malignant progression. Mechanistically, IL-38 is dispensable for epidermal mutagenesis. IL-38 keratinocyte-specific deletion reduced proliferative gene expression along with epidermal cell proliferation and hyperplasia. Additionally, keratinocyte-specific deletion of IL-38 was associated with reduced secretion of inflammatory cytokines leading to a decreased infiltration of myeloid cells into the local tumor microenvironment, these inflammation-driven tumorigenesis are essential for carcinogenesis process. Furthermore, we first demonstrated that IL-38 activated the c-Jun N-terminal kinase (JNK)/activator protein 1 (AP-1) signal transduction pathway to promote proliferation and migration of tumor cells and expression of cancer-related inflammatory cytokines in an IL-1 receptor (IL-1R)-related protein 2 (IL-1Rrp2)-dependent manner. Together, our findings highlight the role of IL-38 in the regulation of epidermal cell hyperplasia and pro-tumorigenic microenvironment through IL-1Rrp2 /JNK and establish a new defined factor IL-38/ IL-1Rrp2 as therapeutic targets in skin cancer.
Generation of mice with a conditional Il9 allele
Authors: Pajulas Abigail , Kaplan Mark H 
Published: February 2023

Abstract
Interleukin (IL)-9 is a pleotropic cytokine that induces cell-specific functions associated with asthma, with target cells including T cells, mast cells, eosinophils, and epithelial cells. Previous findings have demonstrated that IL-9 promotes mast cell growth and differentiation, suggesting that IL-9 contributes to mast cell-driven allergic diseases. Although studies have demonstrated the impact of IL-9 in allergic asthma and other allergic diseases, the cellular source of IL-9 in each setting remains unclear. IL-9 can be produced by several cell types including natural killer T cells, innate lymphoid cells, mast cells, and CD4+ T helper cells (Th). We speculate that T helper 9 (Th9) cells are a major source of IL-9 and that Th9-dervied IL-9 promotes mast cell expansion, activity, and function. Our hypothesis is supported by findings in patients with asthma in which IL-9 expression and Th9 cell numbers are significantly elevated. Here, we address the role of T cell–derived IL-9 in mice with an inactivation of the IL-9 gene restricted to T cells generated by CD4-Cre/loxP-mediated targeting of the IL-9 gene. Preliminary results demonstrate efficient deletion of Il9 in T cells. Il9 fl/fl CD4-Cre Th9 cultures have minimal production of IL-9. Ongoing studies will directly examine the contribution and function of TH9-derived IL-9 during models of allergic airway inflammation and future studies will define the requirement for IL-9 in other cell populations during allergic immune responses.
AKST4290, a CCR3 Inhibitor, Reduces Age-Related Inflammation and Immune Cell Infiltration
Authors: Teichert Arnaud , Masumi Juliet , Rege Sanket , Braithwaite Steven , Minami Sakura 
Published: February 2023

Abstract
Eotaxin, a potent pro-inflammatory mediator, is a chemokine whose levels in plasma increase with age. CCR3, the primary receptor for eotaxin, is expressed on immune cells such as eosinophils and subsets of T lymphocytes, making it a potential therapeutic target for age-related inflammation. Using the CCR3 selective antagonist AKST4290 we aimed to assess the effects of modulating eotaxin/CCR3 signaling in mouse models of aging, inflammation and peripheral immune cell infiltration.
In an aging model, using 18-month-old C57BL/6 mice, AKST4290 treatment reverted the increased blood levels of immune cells to those seen in young mice, with no effects on immune cell levels in young mice.
In inflammation models, using either 8-week-old SKH1-Elite mice challenged with Oxazolone or 8-week-old C57BL/6 mice challenged with LPS, AKST4290 reverted the increased blood levels of immune cells to baseline levels and did not affect cell types unchanged by the stressors. In contrast, Dexamethasone produced a broad, non-specific, decrease of total white blood cells and critical subpopulations.
In an immune cell infiltration model, using 8-week-old C57BL/6 mice challenged with experimental autoimmune encephalomyelitis (EAE), AKST4290 significantly decreased EAE-induced T cell infiltration and microgliosis in the brain.
These findings suggest that AKST4290 is a potent immunomodulator that normalizes immune cell populations related to inflammation to healthy levels, both in the context of disease and aging. These immune modulatory and anti-inflammatory effects have functional consequences in aging models, indicating its therapeutic potential in multiple age-related disorders.
Detection of Cardiotrophin-like Cytokine Factor 1 (CLCF1) by flow cytometry
Authors: Laplante Véronique , Nadeau Ulysse , Rousseau Marine , Lesage Sylvie , Gauchat Jean-François , Pasquin Sarah 
Published: February 2023

Abstract
Cardiotrophin-like Cytokine Factor 1 (CLCF1) belongs to the IL6 family of cytokines and possesses pro-neurotrophic and immuno-modulating functions. Coding mRNA for CLCF1 has been detected in primary and secondary lymphoid organs (i.e. lymph nodes, spleen and bone marrow), as well as in the lungs and feminine reproductive organs. Modulation of CLCF1’s mRNA levels has been associated with the Th17 polarization in CD4+ T cells. However, little information is available regarding CLCF1 protein levels in these tissues or the nature of the immune cells responsible for its production. This can be explained by a lack of in situ detection options for CLCF1. We have therefore developed a methodology for the detection of human and murine CLCF1 by flow cytometry in permeabilized cells. This technique has been validated using derivatives of the Ba/F3 cell line in which cDNAs coding for human and murine CLCF1 were introduced by transduction with recombinant retroviruses. We are currently using this approach to study CLCF1 production by human and murine immune cells. Preliminary results indicate a production of CLCF1 by Th17 T cells. The development of a method to detect CLCF1 by flow cytometry will be beneficial for the study of CLCF1’s functions in the regulation of the immune response.
Functional characterization of gamma-delta (γδ) T cells in allergy
Authors: Yu Esther Dawen , Wang Eric , Sutherland Aaron , Westernberg Luise , Frazier April , Peters Bjoern , Sette Alessandro , da Silva Antunes Ricardo 
Published: February 2023

Abstract
Despite a growing consensus on the involvement of γδ T cells in allergy and other human immunological disorders, the detailed mechanisms remain hypothetical due to lack of investigative tools. Herein, we sought to develop functional assays to study the role of γδ T cells in allergy. Using an Activation-Induced Marker (AIM) assay, based on the upregulation of 4-1BB (CD137) and CD69, we were able to detect ex vivo allergen-specific responses from γδ T cells to multiple allergen extracts in human PBMCs, including mouse, cockroach (CR), house dust mite (HDM) and timothy grass (TG) allergens, which were reproducible and observed in multiple allergic cohorts. The magnitude of allergen-reactive γδ T cells differed between allergic and non-allergic cohorts with HDM and TG allergy, but not mouse or CR allergy. Further characterization showed that the γδ T cells reactive to mouse allergen were mostly the Vδ2 subset and from effector memory (Tem and Temra) compartments. Interestingly, in mouse-specific allergic donors we found differential polarized responses for γδ and ab T cells, with a Th1 and Th2 pattern, respectively. Using a complementary AIM assay for CD40L (CD154) combined with intracellular staining (ICS), we observed that Th1 polarization (INFg and TNFa) in responses to mouse extract was also common in non-allergic donors. Conversely, In the case of CR responses, Th1 polarization of Vδ2 T cells was only observed in the allergic cohort. Overall, these results suggest that allergen-specific Vδ2 T cells are potential to skew dysregulated cytokine responses in immunotherapy. In conclusion, we developed new functional assays that serve as tools for in-depth characterization and deciphering the roles of γδ T cells in allergy.
Human and mouse mast cells express surface PrP that is shed upon activation and affects degranulation
Authors: Willows Steven D , Kulka Marianna 
Published: February 2023

Abstract
Misfolded prion protein (PrP) is the causative infectious agent in prion diseases in both humans and animals. Despite this, the physiological role of PrP in its native conformation (PrPC) is still poorly understood. While best known for its presence on neurons, PrPC is also present on various immune cells. We determined whether mast cells also expressed surface PrPC and whether PrPC expression was linked to changes in mast cell function. Mast cells are tissue-resident immune cells that modulate many physiologic responses such as allergic inflammation, tissue remodeling, response to infection and neuroplasticity. When activated, mast cells degranulate rapidly, releasing several pre-formed signaling molecules and enzymes. We hypothesized that mast cells expressed PrP and that its expression was influenced by mast cell activation. Human (HMC-1 and LAD2) and rodent (BMMC) mast cell lines expressed surface PrPC with the highest expression in BMMC. Activation of mouse bone marrow derived mast cells (BMMC) with A23187 caused a 12 fold decrease in surface PrP expression after 30 min. Surprisingly, levels of PrP recovered to almost half of untreated levels 3 hrs post-stimulation, and reached similar levels to untreated by 48 hrs. BMMCs derived from PrP knockout mice activated by IgE/antigen released 15–25% less granule contents compared to wild type BMMC, suggesting that PrP is somehow involved in degranulation. There were no significant differences in cell surface expression of the IgE receptor FcɛRI in wild type and knockout BMMC. Our results indicate that PrP is highly expressed on human and rodent mast cells and that it may play an important role in degranulation and activation of mast cells, independent of FcɛRI expression.
Aryl hydrocarbon receptor regulation of T follicular helper cells during respiratory viral infection
Authors: Houser Cassandra L 
Published: February 2023

Abstract
The aryl hydrocarbon receptor (AHR) is a ligand-regulated transcription factor that binds structurally diverse molecules from the environment, including pollutants, dietary factors, and chemicals from microorganisms. AHR-binding compounds alter adaptive immune responses, but the cellular components affected and governing mechanisms are not fully defined. For instance, AHR activation affects antibody responses; a process for which T follicular helper cells (Tfh cells) are critical. We recently discovered that AHR activation alters the percentage and number of Tfh cells during influenza A virus (IAV) infection. Yet, how the AHR modulates Tfh cells is not known. The present work shows that AHR-mediated changes to Tfh cells are due to changes in CD4+ T cell proliferation and BCL6 expression, an essential transcription factor for Tfh cell differentiation. Utilizing conditional AHR knockout mice, we show that changes to Tfh cells require AHR expression in CD4+ T cells. The AHR regulates gene expression directly, via its DNA-binding domain (DBD), and indirectly via other signaling molecules. To delineate which pathways it uses to modulate Tfh cells, we used mice with a mutated AHR DBD and demonstrate that it requires its cognate DBD to regulate Tfh cells during infection. This supports that AHR modulates Tfh cells by directly regulating gene expression. Overall, these findings provide new information about how AHR signaling influences humoral immunity to a common respiratory pathogen. Given that exposure to AHR-binding pollutants impacts immune responses to infections and vaccines, better understanding of the mechanisms that control Tfh cell responses has broad reaching impact on immune defenses and immune-mediated diseases.