bluuebunny/crossref_metadata_2025_split huggingface dataset:
Activation of cannabinoid receptor type-1 (CB1) and type-2 (CB2) inhibits the release of vascular endothelial growth factor (VEGF-A) from human neutrophils
Authors: Braile Mariantonia , Scala Sara , Modestino Luca , Cristinziano Leonardo , Marcella Simone , Ferraro Anne Lise , Galdiero Maria Rosaria , Loffredo Stefania 
Published: January 2023

Abstract

Background
Neutrophils (PMNs) are innate immune cells with primary roles in the acute phase of inflammation and resistance against invading pathogens. Activation of cannabinoid receptor type-1 (CB1) and −2 (CB2) has been suggested as a new strategy to modulate angiogenesis in vitro and in vivo. Angiogenesis, the formation of new blood vessels is complex processes that require the coordinated action of several factors, called vascular endothelial growth factors (VEGFs) and angiopoietins (Ang). These factors are produced by several cells, included PMNs.

Objective
We sought to investigate whether endocannabinoids and their receptors modulate PMNs production of VEGF-A, Ang1, CXCL8 and reactive oxygen species (ROS) production.

Methods
PMNs were isolated from buffy coats of healthy donors. Cells were stimulated with CB1 and CB2 agonists/antagonists alone and in association with LPS. Mediators concentrations in cell-free supernatants were measured using ELISA, whereas ROS production was evaluated by fluorescence assay.

Results
CB1 and CB2 agonists did not induce VEGF-A, Ang1 and CXCL8 release from PMNs but reduced spontaneous release of VEGF-A. LPS activated PMNs causing VEGF-A and CXCL8 release. Interestingly, preincubation of PMNs with agonists inhibited in concentration dependent manner VEGF-A release induced by LPS but did not affected CXCL8 production. The effect of CB agonists of LPS induced VEGF-A was reverted by the treatment with CB antagonists. Finally, CB agonists and antagonists promoted ROS production.

Conclusions
Activation of CB1 and CB2 agonists on PMNs might be a novel strategy to modulate immune cell-assisted angiogenesis.
Irgm1-deficiency leads to myeloid dysfunction in colon lamina propria and susceptibility to the intestinal pathogen Citrobacter rodentium
Authors: Hammer Gianna , Huang Hsin-I , Taylor Gregory 
Published: January 2023

Abstract
IRGM and its mouse orthologue Irgm1 are dynamin-like proteins that regulate vesicular remodeling, intracellular microbial killing and pathogen immunity. IRGM dysfunction is also linked to inflammatory bowel disease (IBD), and while it is thought that defective intracellular killing of microbes underscores IBD susceptibility, studies have yet to address how IRGM/Irgm1 regulates immunity to microbes relevant to intestinal inflammation. Here we find that loss of Irgm1 confers marked susceptibility to Citrobacter rodentium, a noninvasive intestinal pathogen that models inflammatory responses to intestinal bacteria similar to that of IBD. Irgm1-deficient mice fail to control C. rodentium outgrowth in the intestine, leading to systemic pathogen spread and host mortality. Surprisingly, susceptibility due to loss of Irgm1 function was not linked to defective intracellular killing of C. rodentium or exaggerated inflammation, but was instead linked to failure to remodel specific colon lamina propria (C-LP) myeloid cells that expand in response to C. rodentium infection and are essential for C. rodentium immunity. Defective immune remodeling was most striking in C-LP monocytes, which were successfully recruited to the infected C-LP, but subsequently underwent apoptosis. Apoptotic susceptibility was induced by C. rodentium infection and was specific to this setting of pathogen infection, and was not apparent in other settings of intestinal inflammation. These studies reveal a novel role for Irgm1 in host defense and suggest that deficiencies in survival and remodeling of C-LP myeloid cells that control inflammatory intestinal bacteria may underpin IBD pathogenesis linked to IRGM dysfunction.
TLR5 and TLR7 amplify different stage of myeloid cells
Authors: Jung Minho , Choi Eun Young 
Published: January 2023

Abstract
Balance between immune activation and immune suppression is important for immune homeostasis. Toll-like receptors (TLRs) play key roles in the innate immune system, initiating inflammatory responses against pathogen infections and internal danger signals. Different TLRs recognize different kinds of ligands, but not all the TLRs do have same effect: some TLRs are immune stimulatory, while the others are immune suppressive. For instance, TLR5 ligand has been known to be immune suppressive, while TLR7 ligand has been used for immune activation. To understand how these TLR ligands lead to opposite immune outcomes, we investigated the effects of the ligands on myeloid cell differentiation. Treatment of TLR5 ligand flagellin resulted in the increase of CD11b+ Ly6C+Ly6G+ myeloid-derived suppressor cells (MDSCs) on days 2 and 3 after culture initiation. However, TLR7 ligand gardiquimod led to enhancement of dendritic cell differentiation on day 1, and then increase of CD11c− MHCII− CD11b− Ly6C− Ly6G− progenitor population on day 2, which differentiate into CD11c− MHCIIow CD11b+ Ly6C+ cells with DC differentiation potential afterwards. Based on these results, we suggest that different immune outcomes of TLR ligands would depend on the stages of myeloid cells at which corresponding TLRs act, contributing to overall immune homeostasis.
Repeated malaria exposures associate with skewing of monocytes/macrophages toward a regulatory phenotype
Authors: Guha Rajan , Arora Gunjan , Li Shanping , Doumtabe Didier , Doumbo Ogobara k , Doumbo Safiatou , Kayentao Kassoum , Ongoiba Aissata , Traore Boubacar , Crompton Peter D 
Published: January 2023

Abstract
In malaria-naïve individuals, P. falciparum (Pf) infection results in numerous Pf-infected red blood cells (iRBCs) that trigger systemic inflammation and fever. Conversely, repeatedly infected individuals in endemic areas are often asymptomatic and have low levels of iRBCs, even children who have yet to acquire reliably protective antibodies. The molecular mechanisms underlying these clinical observations are unclear. PBMCs collected from Malian children before the malaria season responded to iRBCs by producing pyrogenic, pro-inflammatory mediators such as IL-1β, IL-6 and IL-8. However, following febrile malaria there was a marked shift in the response to iRBCs with the same children’s PBMCs producing lower levels of those cytokines. These data suggest that malaria-induced epigenetic reprogramming of innate immune cells may play a role in immunity to malaria. Accordingly, age-stratified analysis of monocytes collected before the malaria season showed an inverse relationship between age and pro-inflammatory cytokine production capacity. Monocytes of Malian adults expressed higher levels of CD163 and arginase1, associated with a regulatory phenotype. These observations were recapitulated with an in vitro system of monocyte-macrophage differentiation whereby re-exposure to iRBCs was associated with diminished expression of pro-inflammatory mediators and a corresponding decrease in epigenetic markers of active gene transcription (i.e. H3K4me3) at the TSS of the same pro-inflammatory mediators. Together these data support the hypothesis that epigenetic reprogramming of monocytes/macrophages toward a regulatory phenotype contributes to clinical immunity to malaria.
Investigations of Salmonella Motility by the Defensin HD6
Authors: Akahoshi Doug T , Natwick Dean , Collins Sean , Bevins Charles L 
Published: January 2023

Abstract
Antimicrobial peptides are key effector molecules throughout nature responsible in part for the defense against microbial pathogens and influencing the composition of colonizing microbes. Defensins are the major antimicrobial peptides produced by Paneth cells in the mammalian small intestine and are essential for maintaining homeostasis between host and gut microbes. While other defensins have potent microbicidal activity, human Paneth cell alpha defensin 6 (HD6) lacks such activity, yet provides protection from invasion by enteric bacterial pathogens. Using a transgenic mouse model expressing HD6 in Paneth cells, we reported the importance of HD6 in preventing the translocation of pathogenic Salmonella Typhimurium (STM) across the small intestinal epithelium (doi:10.1126/science.1218831); however, the mechanism underlying the inhibition of invasion is not fully understood. We hypothesize that HD6 binds to extracellular bacterial protein structures, such as flagella, and inhibits swimming motility through a process of direct binding and self-assembly. Using a semi-solid agar based motility assay, we demonstrate Salmonella radially migrate from the site of inoculation at a slower rate when in the presence of HD6. In ongoing studies using live epifluorescence microscopy to observe GFP-expressing STM in the presence of HD6, we observe a dramatic decrease in STM motility in a concentration dependent manner with some STM appearing to be completely immobilized, but without changes in bacterial viability. With these findings in mind, we plan to expand our investigation to include both structural variants of HD6 and mutants of STM to more clearly define the key elements of peptide and bacteria that mediate this activity.
Dynamics of migration patterns of polymorphonuclear myeloid-derived suppressor cells during tumor progression
Authors: Fu Shuyu , Patel Sima , Mastio Jerome , Dominguez George A , Torres Kevin Alicea , Nefedova Yulia , Zhou Jie , Gabrilovich Dmitry I 
Published: January 2023

Abstract
Although recruitment of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) to the tumor site is well documented, little is known about their migration patterns within bone marrow in tumor progression. We found that the activation of neutrophils migration in cancer is a two-phase process. The first phase was characterized by the accumulation of a previously uncharacterized population of neutrophils that lack immunosuppressive activity but display a potent ability to spontaneously migrate, whereas the later phase is associated with accumulation of neutrophils with typical features of PMN-MDSC with low migratory activity in the bone marrow of mice with various subcutaneous (ectopic) tumors and in mice with advanced orthotopic lung cancer. Mechanistically, PM-LCs displayed increased metabolic flux through oxidative phosphorylation and glycolysis and have more ATP than that of control neutrophils. However, PMN-MDSC were indistinguishable from neutrophils from tumor-free mice in their metabolic activity. In line with studies of mice, CD15+ neutrophils exhibited greater spontaneous migration than that of PMN-MDSCs from the same patient with cancer. These results elucidate the dynamic changes that neutrophils undergo in cancer.
Focal tumor ablation by irreversible electroporation augments immunotherapy to promote tumor growth control and formation of tumor antigen-specific tissue resident memory CD8+ T cells
Authors: Burbach Brandon J , O’Flanagan Stephen D , Rollins Meagan R , Slaughter Joseph R , Young Katharine M , Song Meredith , Shao Qi , Azarin Samira M , Bischof John C , Shimizu Yoji 
Published: January 2023

Abstract
Memory CD8+ T cells populate non-lymphoid tissues (NLT) and can protect against malignancies, but little is known about the establishment of tumor-specific tissue resident memory CD8+ T cells (TRM) by therapeutically initiated immune responses. We used a transplantable mouse model of prostate carcinoma and MHCI tetramers to track and quantify tumor-specific CD8+ T cells. Tumor challenge led to systemic expansion of endogenous, naïve tumor antigen-specific CD8+ T cells and established a small population of TRM (CD44hi, CD62Llo, CD69+, CD103+) in several NLTs. Previously, we demonstrated that debulking primary tumors using irreversible electroporation (IRE) induced expansion of tumor-specific T cells. We hypothesized that IRE would cooperate with checkpoint blockade to increase antigen-specific TRM and promote tumor clearance in vivo. Combination treatment with IRE followed by α-CTLA4 promoted a robust 50-fold expansion of proliferating (Ki67+) tumor-specific T cells in blood, a 10-fold increase of TRM in NLT, and was associated with complete remission of residual tumor. Conversely, monotherapy allowed tumor outgrowth and failed to boost T cell numbers, resulting in small populations of PD-1hi tumor-specific CD8+ T cells. Similarly, anti-PD-1 alone or following IRE was also ineffective, suggesting that combined IRE/α-CTLA4 therapy targets newly primed T cells. IRE also enhanced antigen presentation by XCR1+ dendritic cells, promoting T cell priming in regional lymph nodes. These results indicate that combining α-CTLA4 checkpoint immunotherapy with focal tumor ablation, such as IRE, capable of releasing high quantities and qualities of endogenous tumor antigens may serve as a potent in situ tumor vaccination strategy.
ITK regulates antigen-specific CD8+ T cell-derived IL-10 production and modulates influenza-induced pulmonary immunopathology
Authors: Huang Weishan , Solouki Sabrina , August Avery 
Published: January 2023

Abstract
Influenza (flu) infection is a leading cause of respiratory disease and death worldwide, causing 3-5 million cases of severe illness and more than 250,000 deaths during an average flu season. Severe life-threatening influenza infections in humans are accompanied by an aggressive pro-inflammatory response and an insufficient anti-inflammatory immunity. The production of the immunosuppressive cytokine IL-10 by virus-specific T cells is critical in limiting the immunopathology during flu infection, however, our understanding of how this immunomodulatory event is regulated at the molecular level is very limited. Using mouse models that report the production of IL-10 through green fluorescent protein, OTI CD8+ T cells and influenza A viruses that express the OTI antigen ovalbumin (OVA), we find that the T cell receptor (TCR) responsive tyrosine kinase, Interleukin-2 Inducible T cell kinase (ITK), is a critical regulator of the ability of CD8+ T cells to develop into IL-10-producing cells in an antigen specific manner. The absence of ITK resulted in increased morbidity and mortality in murine model post influenza A infection, accompanied by a severe impairment of IL-10 production by CD8+ T cells in the airway. ITK kinase activity is required for its role in promoting IL-10-producing CD8+ T cell differentiation, by regulating the levels of expression of transcription factors including IRF4, Blimp1, AHR and cMAF. Our data suggest that ITK regulates CD8+ T cell production of IL-10 during influenza A infection, and modulates the development of pulmonary immunopathology. Targeting ITK signaling maybe a promising strategy in modulating airway inflammation during viral infections.
Herpesvirus entry mediator promotes Th1 responses in the cornea during ocular HSV-1 infection
Authors: Park Seo Jin , Riccio Rachel , Kopp Sarah , Longnecker Richard 
Published: January 2023

Abstract
Herpes simplex virus-1 (HSV-1) is a major cause of infectious blindness in the developed world. Ocular herpetic disease is characterized by persistent inflammation in the corneal tissue even after the virus has successfully been cleared, which causes irreversible damage to the cornea. It is unknown what molecular mechanisms drive chronic inflammation following ocular HSV-1 infection. Herpesvirus entry mediator (HVEM), a TNFR superfamily member, mediates both stimulatory and inhibitory signals in T cells by interacting with its functional ligands, LIGHT and B and T lymphocyte attenuator (BTLA). Mice lacking HVEM exhibit a significant decrease in Th1 cells, PMNs, and monocytes in the cornea 14 days post-intracorneal HSV-1 infection as well as a decrease in Th1 and Th17 cells in the draining lymph nodes. We hypothesize that HVEM contributes to exacerbation of ocular disease by stimulatory co-signaling through interaction with its ligands. We observe that LIGHT deficient (LIGHT−/−) and BTLA deficient (BTLA−/−) mice alone do not show significant difference in infiltrating immune cells or corneal pathology compared to wild type (WT) mice. Interestingly, mice lacking both BTLA and LIGHT (BTLA−/−LIGHT−/−) exhibit significant reduction in Th1 cells and improvement in corneal pathology. We show that stimulatory HVEM signaling mediated by both LIGHT and BTLA promotes chronic inflammation during ocular HSV-1 infection.
Measurement of MHC-peptide interaction through cell-surface expression and stability assay improves screening of potential CD4+ T-cell epitopes
Authors: Miyadera Hiroko , Nagai Hideaki , Yoshiyama Takashi , Tokunaga Katsushi , Hoshino Yoshihiko 
Published: January 2023

Abstract

Introduction
Accurate prediction of T-cell epitopes have remained a bottleneck for developing peptide vaccines for infections and cancer immunotherapy, especially for major histocompatibility complex class II (MHC II). We previously found that cell-surface expression level of the covalently linked peptide-MHC II complex is a good indicator of MHC-peptide interaction (Miyadera et al. 2015 J Clin Invest). We named this assay as “delta MHC (dMHC) assay” and validated its utility in the prediction of CD4+ T-cell epitopes.

Method
dMHC assay was performed using engineered fibroblast cells that stably express DRA- or DQA1. The cell line was transduced with pMXs-IG (Kitamura et al. 2003 Exp Hematol) that carries DRB1/3/4/5- or DQB1-peptide fusion construct. Cell surface HLA expression was measured by flow cytometry and normalized to the internal control GFP (Miyadera et al. 2015 J Clin Invest).

Results and Discussion
We conducted systematic binding analysis for HLA class II (27 alleles: HLA-DRB1,3,4,5, and HLA-DQ) and nine peptides derived from Mycobacterium tuberculosis antigens CFP-10 and ESAT-6 that contain potential CD4+ T-cell epitopes (Nagai et al. 2014 J Immunol Res). The cell surface HLA expression level in the presence of the test peptide were normalized to the expression level of the negative control peptide (g9). The normalized expression level (dMHC-g9) showed overall good correlation with the predicted binding affinity (IC50). Moreover, dMHC-g9 outperforms IC50 in predicting potential T-cell epitopes. These data indicate that dMHC assay may be suitable as a platform for large scale analysis of MHC-peptide interaction and data sharing, which should facilitate improved prediction of CD4+ T-cell epitopes.
Histotripsy initiates local and systemic immunological response and reduces tumor burden in breast cancer
Authors: Hendricks Alissa Danielle , Howell Justin , Schmieley Rebecca , Kozlov Shawn , Simon Alex , Coutermarsh-Ott Sheryl L , Vlaisavljevich Eli , Allen Irving C 
Published: January 2023

Abstract
Histotripsy is an emerging, non-invasive, and non-thermal tumor ablation method that has been shown to ablate tissues with millimeter precision guided by real-time ultrasound imaging feedback. Histotripsy ablates tissues by focusing high pressure ultrasound pulses to create cavitation, liquefying tissue. By utilizing this mechanism, preliminary data has suggested that Histotripsy could be used to both ablate a primary tumor and to stimulate the immune system to treat metastatic disease. However, there has not been work done to date on investigating the interactions of the immune system with Histotripsy. For the current work, we hypothesized Histotripsy treatment of a 4T1 tumor in a murine model would stimulate local and systemic immune system responses with an improvement on disease burden. Here, we show that Histotripsy was able to reduce the size of the treated tumor, number of metastases, and change the cytokine profiles. Treated tumors had an average of 15% reduction in volume in 24hrs following treatment and showed increases in necrosis and innate immune cell infiltration. Analysis of early response revealed increased pro-inflammatory cytokines, correlating with increased anti-tumor microenvironments. At a long term follow up, Histotripsy decreased disease burden through primary tumor size and metastasis number. These results demonstrate Histotripsy can stimulate an immunological effect with the increased anti-tumor microenvironment in the treated area one day after treatment, and overall disease morbidity in the long term. Histotripsy has promise as an immunomodulating tumor ablation method for breast cancer, and, once established, has the potential to be utilized in different tumor types as well.
Weighted Gene Co-expression Network Analysis of CXCL13, CXCR5, and associated genes in multiple myeloma
Authors: Adebayo Olayinka Omoyitola , Young Corey D , Carey Kaylin , Dill Courtney D. , Nunez Sha’Kayla K , Griffen Tiara L , Jain Sanjay , Lillard James W 
Published: January 2023

Abstract
Multiple myeloma (MM) is characterized by the neoplastic proliferation of plasma cells. In 2018, there were approximately 30,770 new cases diagnosed and about 12,770 dead. Characterizing its molecular phenotypes is important for effective treatment of MM and predicting relapse. CXCL13-CXCR5 interactions are involved in malignancy cell homing, adhesion, signal transduction, and calcium flux, all of which promote MM progression. Here, we analyzed RNA-sequence data from matched normal, primary and relapsed MM cases. Patient control mRNA samples matched primary tumor and relapse samples were acquired from the database of Genotypes and Phenotypes to evaluate the mRNA expression patterns of CXCL13, CXCR5 and associated genes in MM. Bioinformatics tools were to integrate published genomic data from MM patients (n=480) and identify genes associated with CXCL13-CXCR5 signaling. DESeq analysis was used to determine differentially expressed genes between normal tissues, primary tumor, and relapse groups. Weighted gene co-expression network analysis identified clusters of genes significantly associated with the molecular phenotypes of MM. Notably, gene expression is driven by NFAT and JUN, known to be activated by the CXCL13-CXCR5 axis, and plasma cell signaling pathways significantly correlated with select MM molecular phenotypes and patient survival. Ingenuity pathway analysis was performed to analyze upstream regulators, gene interaction and canonical pathways. Taken together, our data show CXCR5-CXCL13 signaling networks are significantly expressed and associated with MM pathogenesis and plasma clonality. This study provides a better understanding of the heterogeneous nature of MM and the role of CXCL13-CXCR5 axis in MM.