bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hyper MAPK/ERK signaling in DCs causes increased LPS-induced TNFa at the post-transcriptional level
Authors: Minichino Danielle E , Behrens Edward 
Published: February 2023

Abstract

Background:
Langerhans Cell Histiocytosis (LCH) is an inflammatory disease driven by abnormal dendritic cells (DCs) with hyper-MAPK/ERK signaling, usually due to Braf-V600E mutation. Since DCs are poised with Toll like Receptors (TLRs), which utilize MAPK/ERK signaling to incite an immune response, we hypothesize that LCH cells have a hyper-TLR response.

Methods:
We use an animal model of LCH (CD11c-Cre:BRAFV600E-flox; which expresses Braf-V600E in DCs) to measure serum TNFa and tnfa transcripts in CD11c+ splenocytes after LPS injection. Cultured BMDCs were pre-treated with/without a V600E-inhibitor before LPS stimulation. We measured intracellular TNFa, secretion and reuptake of TNFa, tnfa transcripts, NFkB signaling, TNFa degradation, and TACE activity.

Results:
LCH mice have increased LPS-induced circulating TNFa compared to WT. LCH-BMDCs have reversible LPS-induced increase in TNFa secretion and intracellular accumulation, but decreased tnfa and tlr4 transcripts and NFkB signaling compared to WT. There is no difference in TACE activity, TNFa reuptake, or protein degradation.

Conclusions:
Hyper-MAPK/ERK signaling in DCs causes a hyper LPS-induced TNFa response, despite diminished LPS signaling. Increased intracellular TNFa indicates a cell intrinsic, post-transcriptional mechanism. Current experiments are analyzing translation of TNFa, which is implicated by MAPK/ERK signaling. These data indicate novel effects of Braf-V600E on LPS-induced TNFa production, and current studies are investigating whether this is a more global phenomenon.
Distinct chromatin modifications at the Il33 locus in mast cells contribute to sex dimorphic susceptibility to EAE, an autoimmune CNS demyelinating disease
Authors: Brown Melissa Ann , Yang Yuchen 
Published: February 2023

Abstract
Many autoimmune diseases, including multiple sclerosis (MS), exhibit a striking female bias, but the underlying mechanisms remain poorly understood. Using the SJL mouse model of MS, experimental autoimmune encephalomyelitis (EAE), which recapitulates this sex dimorphism, we previously demonstrated that preferential IL-33 expression contributes to male protection. IL-33 expression by mast cells in PLP139–151-immunized males activates type 2 innate lymphoid cells, which in turn drives a non-pathogenic Th2 response to myelin peptide. Testosterone directly activates Il33 in male but not female mast cells, but this male-specific response is also observed with other modes of activation, suggesting there are constraints on the chromatin landscape that limit Il33 expression in females. Here we use chromatin immunoprecipitation (ChIP) and ATAC-seq to examine chromatin modifications at the Il33 locus. At steady state, ChIP assays reveal activating H3K4me3 and H3K9ac histone modifications at the promoter and conserved non-coding sequences within the Il33 gene are most prevalent in male-derived cells, while H3K27me3 repressive marks dominate in females. Increased basal Il33 chromatin accessibility assessed by ATAC-seq is also evident in males. Together, these results indicate that while testosterone can directly stimulate IL-33 production, it also exerts effects on Il33 during development conferring a higher potential for expression in males by altering the chromatin landscape. RNAseq analyses of IgE-activated mast cells revealed a broad range of sex-biased gene expression, indicating that sex hormone influences during development drive epigenetic changes and may be a common mechanism to explain sex dimorphism in immunity.
Mixed Lineage Kinase 3 Promotes Dexamethasone-Induced Thymocyte Apoptosis
Authors: Kumar Sandeep , Singh Sunil Kumar , Viswakarma Navin , Sondarva Gautam , Nair Rakesh Sathish , Rana Basabi , Rana Ajay 
Published: February 2023

Abstract
Dexamethasone (DEX) is a corticosteroid known to regulate T-lymphocyte growth/differentiation and used in the clinic as an anti-inflammatory and immunosuppressant agent. The glucocorticoid hormones, including DEX, can induce thymocyte apoptosis. It has been reported that DEX-induced thymocyte apoptosis is associated with an early stage ceramide generation. Since we reported earlier that ceramide induces Mixed Lineage Kinase 3 (MLK3) activation and therefore, we probed whether MLK3 inhibition could attenuate DEX-induced thymocyte apoptosis. Here, we report that genetic loss of MLK3, a member of MAP3K family attenuates DEX-induced apoptosis in thymocytes. To understand any differences in thymic T cell surface markers at the basal level between WT and MLK3−/− mice, we determined protein expression of various surface proteins involved in T cell activation and apoptosis. Flow cytometry analyses indicated that the genetic loss of MLK3 increases CD27 expression on thymic CD4+, CD8+, and CD4+CD8+ T cells; however, CD28 expression was mainly increased on CD8+ and CD4+CD8+ T cells due to loss of MLK3. Interestingly, genetic loss of MLK3 paradoxically regulated CD70 expression on thymic and splenic T cells. Furthermore, 4–5 weeks old WT and MLK3−/− mice were challenged with DEX for 24 h, and apoptosis in thymic T cells was determined. The flow cytometry results showed a significant decrease in the DEX-induced apoptosis in thymic CD4+, CD8+, and CD4+CD8+ T cells, isolated from MLK3−/− mice. Taken together, these results suggest that MLK3 could be a potential target for DEX-induced thymocyte apoptosis and its inhibitors can be used to mitigate thymocyte apoptosis.
Highly protective T-h17-polarized responses against Influenza A Virus develop in the absence of CD4 T cell-intrinsic T-bet and Eomes expression
Authors: Dhume Kunal , Finn Caroline M , McKinstry Karl Kai 
Published: February 2023

Abstract
Though bulk CD4 T cell responses against influenza A virus (IAV) are characterized as Th1, expression of the transcription factor T-bet (Tbx21), the ‘master regulator’ of Th1 fate, is not required for mice to clear IAV. While the Tbx21−/− CD4 T cells develop some Th17 characteristics in the infected lung, they still retain relatively high Th1 functionality including robust IFNγ production. Here, we show that the transcription factor Eomesodermin (Eomes) is required for the residual Th1 identity of Tbx21−/− cells by comparing Tbx21−/− and double Tbx21−/−Eomes− /− CD4 T cells in an adoptive transfer model of IAV infection. The Tbx21−/−Eomes−/− cells not only lose Th1 attributes but develop much stronger Th17 attributes versus Tbx21−/− cells. The elaboration of Th17 responses by Tbx21−/− Eomes−/− cells is not a default effector state but requires receipt of both IL-6 and TGFβ in IAV-infected lungs. Similar Th17 identity in the absence of Th1 attributes are seen analyzing CD4 T cells in Tbx21−/−Eomes−/− mice infected with IAV, and this response correlates with protection as CD4 T cell depletion abrogates viral control. To more stringently test the protective capacity of Th17 responses, we transferred in-vitro Th17 primed WT or Tbx21−/−Eomes−/− effectors to unprimed WT hosts and challenged with lethal IAV. Both populations protected, but while WT Th17 cells gained Th1 characteristics in vivo, Tbx21−/−Eomes−/− cells strengthened their Th17 functionality remarkably without any shift towards Th1 identity. Our observations thus show that prototypical Th17 cells, that cannot gain Th1 attributes through mechanisms of plasticity often seen during pathogen responses, can mediate strong protection against IAV infection.
Helicobacter pylori infected gastric epithelial cells drive dendritic cells towards a tolerogenic phenotype.
Authors: Hashimi Marziah , Schearer Julia , Sebrell Thomas , Gentry Andrew B , Bimczok Diane 
Published: February 2023

Abstract
The interactions of dendritic cells (DCs) with the gastric epithelium are thought to play a role in shaping DC function and thus the adaptive immune response to Helicobacter pylori (H. pylori) infection. We have shown that DCs are recruited to the H. pylori-infected gastric epithelium, positioning them for uptake of bacterial antigen from the gastric lumen. Using a gastric organoid model of H. pylori infection, we sought to define how the interactions between gastric epithelial cells and DCs regulate the antigen-presenting cell capacity of DCs. Supernatants were collected from organoids microinjected with H. pylori (infected) or PBS (mock), and monocyte-derived dendritic cells (MoDCs) were incubated with these supernatants. Flow cytometry data showed that the supernatant from infected organoids drove the DCs toward a tolerogenic state, as the expression of CD40, CD83, and CD86 was significantly decreased compared to incubation with supernatants from mock infected organoids, whereas CCR7 expression was increased consistent with a more tolerogenic phenotype. DCs treated with supernatants from infected, but not mock-infected organoids suppressed the proliferation of naïve autologous T cells. We also assessed T cell polarization using a cytokine array and found an increase in Th1, Th17 and Treg cytokines, but a decrease in Th2 cytokines in T cells that were challenged with DCs pre-treated with supernatants from infected organoids compared to mock-infected organoids. In summary, we have shown that H. pylori-infected gastric epithelium drives the DCs toward a tolerogenic state, resulting in reduced expression of maturation markers and CD4+ T cell proliferation while maintaining the DCs’ ability to induce T cell cytokine secretion.
In vitro expanded human regulatory T cells (tTreg) and CD4+ effector T cells (Teff) display distinct metabolic demands and response to lactate.
Authors: Hippen Keli L , Zhang Yigang , Aguilar Ethan George , Rhee Stephanie , Wagers Sara Bolivar , O‘Connor Roderick , Rathmell Jeffrey C , Blazar Bruce R 
Published: February 2023

Abstract
Thymus-derived regulatory T cells (tTreg) have been used as a cellular therapeutic to prevent or treat autoimmunity, graft rejection, and graft versus host disease (GVHD) in humans. Metabolic pathways affect tTreg differentiation, expansion and suppressive function. Recent experiments challenge the initial paradigm that tTregs depend on fatty acid oxidation (FAO) and glycolysis. Murine studies also show that central memory and effector tTreg have distinct metabolic profiles, but human studies to date use pooled populations of tTreg. The goal of the current study was to identify and compare the metabolic pathways critical for naïve and memory human tTreg, and to determine whether these pathways differ from naïve human CD4 T cells. We found that memory tTreg have higher rates of glycolysis and oxidative phosphorylation (OXPHOS) than naïve tTreg, both at rest and after stimulation. Activated naïve and memory tTreg had high basal OXPHOS, but neither glucose nor FAO were major substrates. Consistent with this, culture in low glucose did not affect expansion of either naïve or memory tTreg, but greatly decreased that of CD4 Teff. Extracellular lactate increased tTreg basal OCR and, in contrast to the lactate-dependent inhibition of CD4 T cells, enhanced tTreg expansion. This is consistent with differential expression of a class of lactate transporters (monocarboxylate transporters, or MCT) where Treg have increased expression of MCT1/2, which favor import whereas Teff have increased expression of MCT4, which favors export. These data demonstrate that naïve and memory tTreg should be analyzed separately, and provide a molecular insight into why tTreg, but not Teff, function in a low glucose, high lactate environment.
Activation of B10 cells via gut microbiota in the pathogenesis of systemic lupus erythematosus
Authors: Prakash Meeta B , Luo Xin M , Mu Qinghui , Edwards Michael R 
Published: February 2023

Abstract
Systemic Lupus Erythematosus(SLE), a disease of various genetic and environmental origins, is often characterized by immune complexes causing inflammation and damage in various organ systems, such as the brain, kidneys, lung, joint, and skin. Current treatment for SLE include steroidal and nonsteroidal anti-inflammatory drugs, antimalarial agents, and immunosuppressive medications. These treatments primarily reduce symptoms, and patients may develop side effects with long-term care. The gut microbiome has an important role in various metabolic and immunological diseases. A previous study found that increased Lactobacillus colonization restored the mucosal barrier, and increased abundance in these bacteria attenuated SLE symptoms through reversing a ‘leaky gut’. This allows for potential therapies of SLE with probiotics. In preliminary trials, vancomycin treatment during active disease killed gram-positive bacteria, but spared Lactobacillus, and attenuated symptoms. Surprisingly, significantly decreased amounts of CD10+CD19+ B cells that produce IL-10 (B10) were found when vancomycin was given before the onset of lupus. Vancomycin treatment before disease onset significantly removed bacterial DNA from the gut and circulation, causing exacerbation of clinical symptoms. Oral gavage of activated B10 cells in vancomycin treated mice rescued the negative effects of vancomycin treated mice at the pre-disease stage. This work suggests that a form of oral tolerance induced by bacterial DNA-mediated expansion of B10 cells suppress disease onset in the autoimmune-prone MRL/lpr mouse model. Future studies are warranted to further define the mechanism behind bacterial DNA promoting B10 cells.
Specific coping strategies in JSLE depression and anxiety – the untold story of brave soldiers
Authors: Poole Brian D , Silva Beatriz , Figueiredo-Braga Margarida 
Published: February 2023

Abstract
Background and aims: Patients with Juvenile-onset Systemic lupus erythematosus (jSLE) have to overcome the burden of unpredictable and disabling disorder. Coping strategies influence patient’s ability to cope with negative emotions, treatment exigencies and physical impairment related with the disease. Our aim was to explore the relationship between coping mechanisms and the presence of anxiety and depression.

Methods:
Thirty jSLE patients answered the Brief Coping Orientation to Problems Experienced (COPE) questionnaire, the Hospital Anxiety and Depression Questionnaire (HADS) and the Subjective Happiness Scale (SHS).

Results:
Clinically relevant anxiety (63.3%) and depressive symptoms (13.3%) were present in the jSLE patients. Our population showed higher coping scores for problem-focused (0,7±0,2), emotion-focused (0,6±0,2) and social supported coping (0,5±0,2), than avoidant coping (0,3± 0,1). Higher depression scores and lower subjective happiness levels correlated with higher anxiety scores (r=0.673, p=0.000 and r=0.592, p=0.001, respectively). Problem focused coping was associated with higher self-reported happiness (r=0.564, p=0.002) and avoidant coping was positively correlated with higher number of anxious symptoms (r=0.560, p=0.002).

Conclusions:
These results underscore the relevance of psychological assessment in jSLE patients. Adaptive coping strategies led to greater happiness, while avoidant coping strategies were associated with higher anxiety. Teaching of adaptive coping mechanisms, in combination with other interventions are likely to enhance the patients’ ability to respond to the multiple physical, social and emotional challenges imposed by this autoimmune disorder.
STING pathway mediated immunologic consequences of DNA damage repair gene losses in high-grade serous ovarian tumors
Authors: Shakfa Noor E. , Lightbody Elizabeth , Li Deyang , Wilson-Sanchez Juliette , Afriyie-Asante Afrakoma , Chenard Stephen , Hamade Ali , Tyryshkin Kathrin , Koti Madhuri 
Published: February 2023

Abstract
High grade serous ovarian carcinoma (HGSC) is the most lethal gynecologic malignancy with high rates of chemotherapy resistance and poor outcomes. Our previous studies have demonstrated the variable tumor immune microenvironment states that associate with platinum chemotherapy response. We further showed the significance of the interferon induced chemokine CXCL10 as a key mediator of tumor infiltrating immune cell recruitment. Using the ID8-Trp53−/− murine model of HGSC, we demonstrated the potential of Stimulator of Interferon Genes (STING) pathway activation in enhancing response of HGSC tumors to carboplatin chemotherapy and sensitizing them to immune checkpoint blockade. Notably, CXCL10 production via IFN1 is also governed by genes that regulate cellular DNA damage repair pathways. Evolving evidence indicates a role of BRCA1 and PTEN genes in mediating cellular IFN1 responses. Losses in the function of these genes is widely prevalent in a large proportion of HGSC tumors, where tumors with BRCA1 mutations have higher CD8+ T cell infiltration in contrast to those with loss of PTEN. We hypothesized that HGSC tumors with loss of PTEN expression can be rendered susceptible to immune mediated killing via activating the STING pathway. Tumors generated from ID8-Trp53−/−; Brca1−/− cells and those from ID8-Trp53−/−; Pten−/− cells in C57BL6 mice showed significant immunologic differences. STING agonist treatment significantly increased chemosensitivity and improved overall response in mice implanted with ID8-Trp53−/−; Pten−/− cells compared to those treated with carboplatin alone, altering immune responses. This study is foundational to guide rationalistic combinations of STING pathway activating therapies in HGSC.
Targeted Chemotherapy and Checkpoint Blockade Mediate Synergistic Protection Against BCR-ABL+ B Cell Leukemia
Authors: Venkatesh Hrishi , Tracy Sean I. , Hekim Can , Harris Lynn-Heltemes , Farrar Michael A. 
Published: February 2023

Abstract
Immunotherapy using anti-PDL1/anti-CTLA4 checkpoint blockade has been successfully employed for cancers with many non-synonymous mutations. Whether checkpoint blockade-based approaches can be adapted to cancers with low mutational burden, such as BCR-ABL+ B cell leukemia, is still unclear. We have previously demonstrated that the endogenous anti-tumor T cell response, although important for initial control of leukemia, is unable to control leukemia by itself. We show here that leukemia induces an exhaustion phenotype in activated T cells, as well as an associated loss of effector function, that could contribute to the inability to clear leukemia. However, checkpoint blockade therapy using aPDL1 and aCTLA4 by themselves had no impact on survival in leukemic mice. Combining aPDL1 blockade with targeted chemotherapy using the BCR-ABL inhibitor nilotinib had a significant synergistic therapeutic effect. The effectiveness of the aPDL1-nilotinib dual therapy was primarily dependent on CD4 T cells, indicating a key role for endogenous anti-tumor CD4 T cells in leukemia clearance. Indeed, a significant proportion of long-term surviving nilotinib + aPDL1-treated mice were protected against a secondary leukemia re-challenge; indicative of an effective memory T cell responses. Thus, combination therapeutic approaches involving checkpoint blockade can elicit potent anti-tumor CD4 T cell responses that induce long-term protection against leukemia. Future studies will explore the mechanisms by which CD4 T cells orchestrate protective anti-leukemia responses.
Obese microenvironments attenuate the efficacy of endogenous and CD19-directed CAR T-cells
Authors: Ank Raira Silva , Henry Curtis J. 
Published: February 2023

Abstract

Background:
Hematological malignancies represent the largest class of cancers in pediatric patients. Despite successes in treating primary diseases, patients with relapsed B-cell acute lymphoblastic leukemia (B-ALL) and diffuse large B-cell lymphoma (DLBCL) have significantly lower survival outcomes. This has led to the use of immunotherapies, notably chimeric antigen receptor (CAR) T-cells, to treat relapse. Unfortunately, only 50% respond to CAR-T therapy, with relapse common in less than a year.

Objective:
Given that obesity is associated with poorer survival outcomes in pediatric patients with B-cell malignancies and promotes a loss of immune homeostasis, we hypothesize that obesity compromises the function of endogenous and engineered T-cells in B-cell malignancies.

Results:
Using in vitro models of adiposity and diet-induced obesity mouse models to study the impact of obesity on primary T-cell function, we observe defects in proliferation, cytokine production, and the upregulation of cytolytic mediators when cultured in adipocyte but not in bone marrow cell or unconditioned media. The introduction of CD19-direct CARs did not overcome adipocyte-induced defects in T-cell function, which resulted in increased PD-1 surface expression, compromised cytokine production, and defective killing of human CD19-expressing B-ALL cells. Interestingly, human B-ALL cells cultured in adipocyte-secreted factors downregulated CD19, suggesting that adipocytes promote B-ALL pathogenesis by compromising T-cell function and reducing the surface expression of antigens on target cells.

Conclusion:
Our studies are the first to demonstrate the inhibitory impact of the adipocyte secretome on CAR T-cell function in models of B-ALL.
Single-cell mapping of human endometrial cancer reveals tumor-specific instruction of tissue-infiltrating leukocytes
Authors: Tong Huan , Feng Hao , Wan Xiaoping 
Published: February 2023

Abstract

Background:
Endometrial cancer (EC) is a hormone-related carcinoma with increased morbidity among female patients of all backgrounds. For second-and third-line treatment, evaluation of tumor DNA mismatch-repair function by determining microsatellite instability (MSI) status is essential to choose proper targeted therapies. Pembrolizumab, a PD-1 immune checkpoint inhibitor, is an effective agent for patients with high-MSI endometrial cancer. Advanced or recurrent ECs identified as MSS are more sensitive to Pembrolizumab plus Lenvatinib, a receptor tyrosine kinase inhibitor (TKI), which slows the growth of endometrial cancer cells. Unrevealing the complex immune microenvironment of EC and the crosstalk between leukocytes and tumor cells is essential for providing an immunotherapeutic strategy.

Method:
the leukocyte landscape of endometrial carcinoma was mapped using bioinformatic analysis and high-dimensional single-cell profiling (CyTOF), which contained 42 biomarkers as the panel.

Result:
The heterogeneous composition of tissue-resident and invading immune cells within endometrial cancer and adjacent normal tissues permitted a clear distinction. NK cells, monocyte-derived macrophages, and neutrophils were enriched in adjacent normal tissue. A Th9 subtype of T cells with CCR5 and CD38 positive expression was recruited to the invasive margin. PD1 was elevated for these Th9 cells. Intriguingly, PD1+ESRneg T cells, Treg cells, and ESR+ leukocytes such as ESR+ granulocytes and Siglec1+CCR5+CD40+ESRhi macrophage were enriched in the infiltrating immune cells.

Conclusion:
Defining the specific immunological signature of EC can facilitate the rational design of targeted immunotherapy strategies.