bluuebunny/crossref_metadata_2025_split huggingface dataset:
Countering Inflammatory Arthritis by 14-3-3zeta: A Novel Target
Authors: Chakravarti Ritu , Kim Joshua , Chun Krista , Durgala Johnathon , Joe Bina , Holoshitz Joseph , Chattopadhyay Saurabh 
Published: February 2023

Abstract
Inflammatory arthritis (IA) is a common inflammatory joint-destructive condition that affects millions of individuals worldwide. Increased immune dysfunction and bone loss have been observed both in IA affected humans and in animal models. Proinflammatory events responsible for RA pathogenesis have been studied in detail; however, the protective mechanism remains under-explored. Earlier, we reported that 14-3-3zeta (z) is a molecular determinant of skewing T cell polarization and interleukin-17A (IL-17A) signal transduction. Here we demonstrate that 14-3-3z is an endogenous suppressor of IA. Loss of 14-3-3z in a novel CRISPR/Cas9 edited knockout (KO) rat resulted in early-onset and severe arthritis in animal models of inflammatory arthritis. Arthritic 14-3-3z KO animals showed increased bone loss and immune cell infiltration in synovial joints. Rescue experiments to supplement the 14-3-3z by immunization during the pre-symptomatic phase resulted in significant arthritis suppression in both wild-type and 14-3-3z KO rats. Immunization generated a robust increase in 14-3-3z antibody and IL-17A, suppressed several cytokines, including IFN-g, TNF-a, and Cxcl-1, preserved tissue mineral bone density, and trabecular bone volume. Our findings, taken together with the prior report of the molecular function of 14-3-3z as an antigen, indicate that antigenic 14-3-3z is responsible for a previously unrecognized inflammatory arthritis-suppressive mechanism.
A metabolically engineered bacterial vaccine protects against arthritis in mice
Authors: de Figueiredo Paul , Guo Fengguang , Das Jugal , Ding Yufang , Jayaraman Arul , Alaniz Robert Christopher , Song Jianxun 
Published: February 2023

Abstract
Arthritis is an autoimmune disease that adversely affects the quality of life of patients. Treatment options for autoimmunity include the use of chemical and biologic agents that suppress inflammation. However, these can result in patients being vulnerable to opportunistic infections. Adoptive T regulatory (Treg) cell transfer can specifically block auto-inflammatory immune cells in an antigen-specific manner while maintaining an intact immune response to pathogens. However, isolating sufficient Treg cells for adoptive cell transfer therapy is challenging; therefore, improving Treg cell survival and function could have important impacts on adoptive Treg transfer therapy. To explore the use of bacterial vaccines as autoimmune therapies, we engineered an attenuated brucellosis vaccine (Brucella melitensis ΔvjbR Tna) to produce indole, a bacterial metabolite that induces Treg expansion. In the collagen induced arthritis (CIA) murine model, Treg transfer following BmΔvjbR Tna treatment decreased arthritis incidence and score. CD4+FoxP3+ T regulatory cells and CD8+ T cells in spleen and lymph nodes were increased in BmΔvjbR Tna treated CIA mice. BmΔvjbR Tna injection also promoted CD4+ cell proliferation, and increased IL-2 and IFN-g expression. The levels of some pro-inflammatory cytokines (IL-1β and IL-6) were also dramatically reduced in BmΔvjbR Tna infected bone marrow derived macrophages (BMDMs). Taken together, these findings indicate that the indole-producing BmΔvjbR Tna strain increases Treg cell populations and proliferation, suppresses proinflammatory cytokine secretion, and reduces autoimmune responses.
Preclinical development of universal SNAP-CAR T cell therapy
Authors: Ruffo Elisa , Kvorjak Michael , Adams Eric , Lohmueller Jason 
Published: February 2023

Abstract
Chimeric antigen receptors (CARs) are artificial T cell receptors that re-target patients’ T cells to specifically recognize and kill tumor cells. Despite remarkable success of anti-CD19 CAR therapy against refractory B cell leukemias, there are several limitations to CAR T cell therapies including toxicities and antigen-loss leading to relapse. To address these issues, we previously developed a “universal” CAR, SNAP-CAR, for which antigen-specificity is directed by co-administered tumor-targeted antibodies. Instead of directly recognizing a tumor antigen, the SNAP-CAR carries out an enzymatic reaction to fuse with antibodies conjugated to a benzylguanine (BG) tag. Activation and effector functions of SNAP CAR T cells can be re-targeted by antibody-BG conjugates to several antigens including: CD20, CD19, HER2, and EGFR. We are now developing SNAP-CAR T cells for potential clinical translation including optimization of the CAR expression construct and functional characterization in a human tumor xenograft mouse model. SNAP-CAR-T2A-LNGFR was cloned into a gamma-retroviral expression system, yielding a 10-fold greater expression level in primary human T cells compared to our previous lentiviral vector. SNAP-CAR T cells were potently activated and lysed tumor cells in a target antigen-specific manner in vitro. Challenging NSG mice with CD20+ Raji leukemia cells, significant tumor regression was observed with SNAP-CAR T cells + Rituximab-BG and anti-CD20-CAR positive control T cells but not SNAP-CAR T cells without antibody, as compared to untransduced T cells + Rituximab-BG. The SNAP-CAR provides a powerful system to program the antigen-targeting capability of human T cells with promise for cancer therapy.
Intracellular PD-L1 regulates DNA damage checkpoints and suppresses Chk1 and PARP inhibitor synthetic lethality
Authors: Kornepati Anand V , Deng Yilun , Dray Eloise , Murray Clare , Kari Suresh C , Osta Erica , Liu Zexuan , Boyd Jacob , Padron Alvaro , Reyes Ryan , Gupta Harshita B. , Clark Curtis A , Li Rong , Zhao Weixing , Vadlamudi Ratna , Sung Patrick , Curiel Tyler J. 
Published: February 2023

Abstract
Tumor PD-L1 mediates non-canonical, intracellular signals including the DNA damage response (DDR), of unclear significance and mechanisms. We show that genetic PD-L1 depletion (PD-L1KO) destabilized the Chk2 protein, a DNA damage response (DDR) factor, resulting in ATM/Chk2 pathway defects but not ATR/Chk1 in melanoma, bladder, breast, and ovarian cancer. Consistent with ATM/Chk2 defects, PD-L1 KO led to DNA damage (γH2AX) and impaired homologous recombination (HR) (p-RPA32, BRCA1, Rad51 foci). Thus, PD-L1 KO vs control cells were significantly more sensitive to DDR inhibitors (DDRi) against ATR, Chk1, and PARP in vitro and in vivo in NSG mice. Chk2 regulation by PD-L1 was independent of PD-L1 cytoplasmic tail yet required intracellular (vs surface) PD-L1 suggesting physical PD-L1/Chk2 interaction supported by IP and imaging. PD-L1 stabilized Chk2 protein by preventing its lysosomal degradation without altering Chek2 mRNA. αPD-L1 is thought to work by protecting PD-L1 induced anti-tumor T cell suppression via PD-1, but PD-L1 DDR effects were PD-1-independent. Intracellular PD-L1 suppressed DDRi induced cGAS/STING activation by immunoblots and qRT-PCR of type 1 IFN genes. In vivo in WT mice, genetic PD-L1 depletion but not αPD-L1, sensitized highly immunotherapy resistant 4T1 breast cancer to PARPi. Strikingly, PARPi had reduced effect on PD-L1KO tumors in RAG2KO mice despite treating WT mice, indicating a strong immune component to DDRi efficacy. Our work implicates a novel role of intracellular PD-L1 in DDR and tumor immunogenicity and identifies related therapeutic vulnerabilities exposed by intracellular PD-L1 targeting. Surface vs intracellular PD-L1, and specific DDR signals could be treatment response biomarkers.
BATF and IRF4 cooperate to counter exhaustion in tumour-infiltrating CAR T cells
Authors: Seo Hyungseok , Gonzales-Avalos Edahi , Zhang Wade , Lio Chan Wang “Jerry” , Rao Anjana , Hogan Patrick 
Published: February 2023

Abstract
Cooperative interactions among transcription factors are essential for gene transcription. We previously showed that NFAT and AP-1 (Fos-Jun) transcription factors cooperate to promote the effector functions of T cells, but that under conditions where it is unable to cooperate with AP-1, NFAT imposes a negative feedback programme of T cell hyporesponsiveness (“exhaustion”). Here we show that BATF and IRF4 cooperate to counter T cell exhaustion. Overexpression of Batf in CD8+ T cells expressing a chimeric antigen receptor (CAR) promoted the survival and expansion of tumour-infiltrating CAR T cells, increased their production of effector cytokines, decreased their expression of inhibitory receptors and the exhaustion-associated transcription factor TOX, and led to the generation of long-lived memory T cells that controlled tumour recurrence. These responses were dependent on the BATF-IRF interaction, since cells expressing a Batf mutant unable to interact with Irf4 did not survive in tumours and did not effectively delay tumour growth. We suggest that BATF overexpression is a therapeutically viable option for improving the anti-tumour responses of CAR TILs, by skewing their phenotypes and transcriptional profiles away from exhaustion and towards increased effector function.
Evaluating the Potential of Harnessing Anti-Leukemia T cells for Treating T-ALL
Authors: Triplett Todd A , Rios Joshua , Kottapalli Srividya , Weinberg Andrew D , Somma Alexander , Sandoval Trevor 
Published: February 2023

Abstract
T cell Acute Lymphoblastic Leukemia (T-ALL) is a devastating malignancy found primarily in pediatric populations. Although patient outcome has improved, standard of care for T-ALL has not progressed from intensive regimens of chemotherapy. Immunotherapy may offer an alternative approach for treating T-ALL with lower toxicity profiles. In the case of B-ALL, adoptive transfer of engineered cells (e.g. CAR cells) has had resounding success. Unfortunately this may not translate to T-ALL due to severe immunodeficiency from ablating the T cell compartment. These studies therefore sought to determine the therapeutic potential of harnessing anti-leukemia T cells that spontaneously arise in tumor-bearing hosts in response to T-ALL. As very little is known about the interplay between T-ALL and T cells, we initially examined changes to the T cell compartment in response to T-ALL during disease progression by transplanting primary leukemia cells into immune-competent, congenic mice. This model revealed an increase in effector memory CD8 T cells with increased levels of PD1 and OX40 that largely correlated with tumor burden. Importantly, co-targeting PD1 with OX40 resulted in drastic reduction in tumor burden while expanding actively proliferating CD8+GrzB+ T cells. To gain a better understanding of T cell interactions with T-ALL in situ, we have begun utilizing NanoString Technologies’ Digital Spatial Profiling platform. This platform has already shown distinct functional states of T cells between those interacting with leukemia compared with those at distal sights. Ongoing studies aim to identify therapeutic targets in multiple organs which can activate T cell populations of interest to maximize their anti-leukemia effector responses.
Diversity of draining lymph node-tumor overlapping T cell receptor repertoire reflects the anti-tumor effect of cancer immunotherapies
Authors: Aoki Hiroyasu , Ueha Satoshi , Tsunoda Mikiya , Ogiwara Haru , Shimizu Haruka , Shichino Shigeyuki , Matsushima Kouji 
Published: February 2023

Abstract
CD8+ tumor-infiltrating lymphocytes, the main effector cells of anti-tumor immune responses, are composed of various T cell clones. Numerous studies have demonstrated the presence of tumor-reactive T cell clones with high frequency in tumors. However, it remains unclear as to which of the two mechanisms, the expansion of limited tumor-specific clones or the induction of diverse tumor-reactive ones, is more beneficial for the action of immune checkpoint inhibitors (ICIs).
To address this question, we performed a T cell receptor repertoire analysis on B16F10 and on Lewis lung carcinoma tumor-bearing mice treated with immunotherapy including anti-PD-L1 monoclonal antibody (mAb) and anti-CD4 depletion mAb, which enhance the expansion of tumor-reactive CD8+ T cells. We focused on the T cell clones that overlapped between the tumor and draining lymph node (dLN) to analyze the T cell clones that were mobilized into the anti-tumor response after treatment.
In anti-PD-L1 and anti-CD4 treated mice, the total frequency of CD8+ dLN-tumor overlapping clones in the tumor was increased. Surprisingly, although the expansion of the top10 overlapping clones did not differ, ICI treatment increased the total frequency of the 11th or lower overlapping clones, indicating the broadening of the tumor-reactive T cell repertoire. Moreover, the administration of FTY720 to tumor-bearing mice, which inhibits egress of T cells from the LNs to circulation, reduced the total frequency of the 11th or lower overlapping clones in the tumor and reduced the long-term anti-tumor effect of anti-PD-L1 treatment.
These results suggest that mobilization of diverse tumor-reactive clones is essential for enhancing and persistent anti-tumor effect of ICIs.
Deep characterization of tumor infiltrating leukocytes using a combination of flow cytometry and single-cell multiomics
Authors: Shi Xiaoshan , Nakamoto Margaret , Middlebrook Aaron , Huang Wei , Lo Evelyn , Neuhoefer Patrick Thomas , Bornheimer Scott , Ghanekar Smita 
Published: February 2023

Abstract
Cancer immunotherapies have the potential to induce durable anti-tumor immune responses; however the efficacy of these treatments varies among patients and partially depends on the complex interplay between malignant cells and non-malignant cells within the tumor microenvironment. In the tumor microenvironment, T cells play a crucial role in eliminating tumor cells and orchestrating anti-tumor immune responses; in contrast immunosuppressive cell types can support tumor progression. Here we harnessed recent advances in single-cell multiomics that allow high-resolution functional characterization of intratumoral immune cells to improve our understanding of their dynamic relationships. Specifically, we utilized a One Solution Workflow to dissociate a solid tumor followed by in-depth dissection of the immune composition of the tumor microenvironment using both flow cytometry and the BD Rhapsody™ Single-Cell Analysis System. We characterized the cellular composition of tumor microenvironment using flow cytometry. Importantly, by examining the expression of mRNA and surface protein markers at single-cell resolution, we unraveled the complexities of the tumor microenvironment, elucidated cellular heterogeneity and identified cell-type specific gene signatures. We propose this One Solution Workflow can be adopted to understand the transcriptional and proteomic phenotypes of tumor infiltrating leukocytes for development of novel immune therapeutic strategies for cancer.
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Inhibition of TH2 responses by a secreted component of a common farm bacterium
Authors: Hollinger Maile Kananiokala , Camacho Daniel F , Hrusch Cara L , Barron Gabriel M , Velez Tania Estefania , Zamora-Pineda Jesus Bernabe , Knight Katherine , Sperling Anne I 
Published: February 2023

Abstract
The rise in incidence of asthma and allergic diseases over the past half century has led to the development of the hygiene hypothesis, which posits that lack of exposure to environmental microbes during childhood leads to inappropriate immune responses to innocuous antigens. Evidence for the hygiene hypothesis is particularly compelling in studies of rural farm children, where infants that live on farms had reduced prevalence of asthma at school age, as compared to their non-farming counterparts. In particular, the presence of exopolysaccharide (EPS) in farm dust independently predicted lower rates of asthma diagnosis. The purpose of this study is to investigate the protective effects of EPS from a common farm microorganism, the hay bacillus B. subtilis, in limiting TH2-type inflammation in a mouse model of asthma. Treatment of house dust mite (HDM)-sensitized mice with EPS was sufficient to block eosinophilia and TH2 accumulation in the lungs, without affecting accumulation of other TH subsets. These effects were dependent on TLR4, the putative receptor for EPS, as EPS no longer protected against eosinophilia and TH2 infiltration in Tlr4−\− mice. Additionally, lung-derived dendritic cells exposed to EPS and HDM in vivo were phenotypically distinct from dendritic cells recovered from mice treated with HDM alone. These data support a protective role for EPS in the inhibition of allergic asthma, potentially by affecting TLR4-dependent dendritic cell priming of TH2 responses.
Single-cell and spatial transcriptomics reveal NLRP3 inflammasome-mediated immunestromal interactions during vasculitis and cardiovascular inflammation.
Authors: Porritt Rebecca A , Zemmour David , Abe Masanori , Chen Shuang , Crother Timothy R , Shimada Kenichi , Arditi Moshe , Rivas Magali Noval 
Published: February 2023

Abstract
NLRP3 activation and IL-1β production are implicated in Kawasaki Disease (KD) pathogenesis, however a detailed description of molecular networks and cellular subsets involved is lacking. Here, in a murine model of KD vasculitis, we used single-cell RNA sequencing and spatial transcriptomics to characterize the cellular landscape of vascular tissues and observed infiltrations of innate and adaptive immune cells, associated with increased expression of Nlrp3 and Il1b. Monocytes, macrophages and dendritic cells were the main sources of IL-1β, whereas fibroblasts and vascular smooth muscle cells (VSMCs) expressed high levels of IL-1 receptor. Genetic inhibition of IL-1β signaling on VSMCs efficiently attenuated the development of cardiovascular lesions during murine KD. In addition, pharmacological inhibition of NLRP3 prevented the development of cardiovascular inflammation. Our results unravel the cellular diversity involved in IL-1β production and signaling in KD cardiovascular lesions and demonstrate that therapeutic strategies targeting NLRP3 might be beneficial for human KD.
Pericardial macrophages and their role in myocardial infarction
Authors: Hughes David M , Won Taejoon , Jurcova Ivana , Talor Monica V , Szarszoi Ondrej , Curnova Lenka , Striz Ilza , Hooper Jody E , Melenovsky Vojtech , Cihakova Daniela 
Published: February 2023

Abstract
Each year an estimated 790,000 people suffer a heart attack in the United States. With such a high incidence, it is important to understand the underlying cellular mechanisms at play during myocardial infarction. In this project we examined the role of pericardial macrophages in modulating the pathogenesis of myocardial infarction. We found that pericardial macrophages expressing the transcription factor GATA6 make up a majority of the cells in the healthy human’s pericardial fluid and have a regulatory function. However, patients with chronic ischemic cardiac disease had their GATA6+ macrophages replaced by inflammatory monocytes in the pericardial fluid. GATA6+ macrophages can be also isolated from murine pericardial fluid. We modeled myocardial infarction through permanent ligation of the left anterior descending artery in adult mice utilizing GATA6+ macrophage deficient LysCreGATA6fl/fl mice. LysCreGATA6fl/fl mice had worse fibrosis and more severe EKG abnormalities compared to control mice following myocardial infarction. Thrombi were also apparent in the hearts of LysCreGATA6fl/fl mice that were not present in control mice. In addition, mice lacking GATA6+ pericardial macrophages had a higher influx of inflammatory Ly6Chi monocytes to the myocardium during the early inflammatory phase of the disease, while pericardial macrophages did not traffic to the myocardium directly at any point. Therefore, we propose that pericardial macrophages inhibit late-stage cardiac remodeling following myocardial infarction by preventing the trafficking of inflammatory monocytes to the site of injury.
Tbet+ B cells facilitate iNKT cell expansion in murine liver and omentum during obesity
Authors: Enslow Benjamin , Vanz Carlo , Schute Travis , Parthasarathy Raksha , Dudley Elizabeth A. , Hägglöf Thomas , Leadbetter Elizabeth A. 
Published: February 2023

Abstract
Obesity is a burgeoning public health crisis, responsible for increasing chronic systemic inflammation which leads to metabolic disease in humans and mice. Understanding the contribution of immune inflammation to metabolic disease is critical for designing effective therapeutic interventions. Invariant Natural Killer T (iNKT) cells are enriched in the adipose tissue, liver, and omenta of both humans and mice, but their frequency is reduced in adipose during obesity. iNKT cells negatively regulate or positively activate other immune cells, including B cells, depending on the immunologic context. Adipose B regulatory cells protect against inflammation, but they are not the only subset of B cells in adipose tissue. Our preliminary studies find increased frequencies of inflammatory Tbet+ B cells in adipose tissue of obese humans and mice which correlates with weight gain in mice and increasing BMI in humans. We now extend those findings to consider iNKT cell interaction with B cells during obesity in the adipose, liver, and omental tissues. Flow cytometry confirmed iNKT cell-dependent expansion of Tbet+ CD11c+ B cells in adipose tissue of obese mice. Interestingly, similar expansion was not observed in liver or omental tissue from the same obese mice. Instead, the livers and omenta of obese mice demonstrated significant increases in iNKT cell frequency compared with lean mice. In contrast to the adipose tissue, the increase in liver and omental iNKT cells during obesity was missing in mice genetically deficient in Tbet+ B cells. Thus, we find that iNKT cell-dependent expansion of Tbet+ CD11c+ B cells within adipose tissue of obese mice is counter-balanced by a reciprocal Tbet+ B cell-dependent expansion of iNKT cells in liver and omentum.
Dectin-1 limits CNS autoimmunity through a non-canonical pathway
Authors: Deerhake M Elizabeth , Danzaki Keiko , Inoue Makoto , Cardakli Emre , Nonaka Toshiaki , Aggarwal Nupur , Barclay Will , Ji Ru-Rong , Shinohara Mari L 
Published: February 2023

Abstract
Innate immunity mediates both damage and repair of the central nervous system (CNS) in neurologic disorders. Although pathologic roles for innate immunity are well-described in multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), protective aspects of the immune response are less understood. In this study, we sought to identify the function of Dectin-1, a C-type lectin receptor (CLR), in CNS autoimmunity. We found that Dectin-1 limited neuroinflammation in EAE while its canonical signaling mediator, Card9, promoted disease. Myeloid cells from the bone-marrow mediated the regulatory function of Dectin-1 in EAE and upregulated expression of Oncostatin M (Osm), a neuroprotective cytokine, through a non-canonical Card9-independent Dectin-1 signaling pathway. We found that the Osm receptor (OsmR) functioned specifically in astrocytes to reduce EAE severity. RNA-seq profiling of the Card9-independent Dectin-1 transcriptional program identified multiple targets with neuroprotective functions, including Osm, which were regulated by NFAT signaling. Our data describe a new mechanism of neuroprotective myeloid-astrocyte crosstalk regulated by a non-canonical Dectin-1 pathway.