bluuebunny/crossref_metadata_2025_split huggingface dataset:
Harnessing the microbiome to regulate systemic innate immunity
Authors: Mihori Saki , Nichols Frank , Provatas Anthony , Blesso Christopher N , Clark Robert B 
Published: August 2023

Abstract
The mechanisms used by the gastrointestinal microbiome to regulate systemic immunity remain unclear. We have identified a microbiome Bacteroidota-derived lipopeptide—L654—that is a TLR2 ligand, accesses the systemic circulation, and thus potentially links the microbiome and systemic innate immunity. We previously documented that Multiple Sclerosis patients have significantly lower serum L654 levels than healthy controls, leading us to postulate that: 1) L654 regulates systemic innate immunity by entering the systemic circulation and mediating TLR2 interactions that maintain “normal” levels of innate immune signaling feedback inhibitors; and 2) inadequate systemic L654 levels result in poorly regulated, enhanced innate immune responses. In prior proof-of-concept studies, we reported that increasing systemic L654 levels by administering exogenous L654 intravenously significantly diminishes systemic innate immune responses and attenuates murine autoimmunity. In the present study, our goal was to document the converse effect, i.e., that decreasing microbiome L654 production enhances systemic innate immune responses. Using wild-type C57BL/6 mice, we report that decreasing microbiome Bacteroidota using specific non-absorbable oral antibiotics reduces fecal and serum L654 levels and significantly enhances innate immune responses. Moreover, the intravenous administration of L654 to antibiotic-treated mice prevents this innate immune enhancement. Overall, our results suggest that L654 is: 1) used by the microbiome to regulate systemic innate immunity; and 2) a manipulatable target capable of enhancing or decreasing innate immunity in the context of infectious, malignant, or autoimmune diseases.
UConn Health Center Academic Advancement Fund
Mucus sialylation determines intestinal host-commensal homeostasis
Authors: Kim Girak , Yao Yikun , Chen Zuojia , Wu Chuan , Lenardo Michael J 
Published: August 2023

Abstract
Intestinal mucus forms the first line of defense against bacterial invasion while providing nutrition to support microbial symbiosis. How the host controls mucus barrier integrity and commensalism is unclear. We show that terminal sialylation of glycans on intestinal mucus by ST6GALNAC1 (ST6), the dominant sialyltransferase specifically expressed in goblet cells and induced by microbial pathogen-associated molecular patterns, is essential for mucus integrity and protecting against excessive bacterial proteolytic degradation. Glycoproteomic profiling and biochemical analysis of ST6 mutations identified in patients show that decreased sialylation causes defective mucus proteins and congenital inflammatory bowel disease (IBD). Mice harboring a patient ST6 mutation have compromised mucus barriers, dysbiosis, and susceptibility to intestinal inflammation. Based on our understanding of the ST6 regulatory network, we show that treatment with sialylated mucin or a Foxo3 inhibitor can ameliorate IBD.
Supported by the Intramural Research Program of the NIAID, NIH, United States and by National Multiple Sclerosis Society Career Transition Award, United States (TA 3059-A-2 to C.W.).
Bi-allelic mutation of CARD11 resulting in fatal immune dysregulation with dramatic B cell expansion
Authors: Markle Janet G , Bhattad Sagar , Gil Deb , Ruchinskas Allison , Snow Andrew L 
Published: August 2023

Abstract
Caspase recruitment domain family member 11 (CARD11) functions as a scaffold to bridge antigen receptors with downstream immune signaling pathways that enable antigen-dependent lymphocyte activation. Given its pleiotropic role in regulating adaptive immunity, different genetic variants in CARD11 can result in diverse phenotypic consequences in humans. Here we describe a patient harboring a novel bi-allelic CARD11 mutation that results in potentially fatal immune dysregulation with massive B cell expansion. He presented with splenomegaly, amenia, leukocytosis and thrombocytopenia within the first year of life. A sibling showed similar symptoms before passing away in infancy. Due to parental consanguinity, an autosomal recessive inborn error of immunity was suspected; whole exome sequencing revealed a homozygous missense mutation in the coiled-coil (CC) domain of CARD11 (p.Arg331Pro, R331P). Functional testing revealed the R331P mutation caused a mild gain-of-function effect relative to wild-type CARD11 in driving constitutive NF-kB activation, suggesting that R331P may disrupt interactions needed to maintain CARD11 autoinhibition in resting lymphocytes. Dramatic B cell lymphocytosis noted in the patient was not present in his heterozygous parents, indicating that 2 mutant alleles are required for this phenotype. In contrast, the patient’s T cells responded poorly to TCR stimulation with severely reduced proliferation. Collectively, these results indicate that the novel R331P CARD11 mutation manifests as the first known recessive form of B cell Expansion with NF-kB and T cell Anergy (BENTA disease), and illustrate how rare monogenic inborn errors of immunity provide a unique discovery platform for basic immunology.
Differential responses to R848 and IgG/IgM in EBV-transformed B cells from individuals with Down syndrome compared to disomic individuals
Authors: James Stephanie , Rosastri Eustacia 
Published: August 2023

Abstract
Down syndrome (Trisomy 21; T21) results from an extra copy of chromosome 21 and causes a myriad of abnormalities, including immune dysfunction. Recent studies of PBMCs in those with T21 have shown reduced numbers of naive B cells, and increases in differentiated B cells, perhaps contributing to increased autoreactive and inflammatory conditions. The receptor for IFNα is located on chromosome 21, and has been shown to be expressed and functional on cells with T21. IFNα has been suggested to play a role in B cell responses, but little is known about the role of IFNα and B cell interaction in those with T21. Herein, we sought to further evaluate B cells from individuals with T21 following in vitro stimulation, with the hypothesis that cells with T21 would have greater B cell activation and cytokine production due to the extra IFNα receptor, compared to their disomic counterparts. EBV-transformed B cells from individuals with and without T21 were cultured and stimulated with R848/IL2, IgG/IgM cocktail or media as a control. In our unstimulated controls our results were in alignment with previous studies that suggest B cells from those with T21 are more terminally differentiated; we also observed significantly increased expression of the activation marker CD86 on these cells compared to disomic individuals. However, when stimulated with either R848 or IgG/IgM we did not observe a significant difference in the ability of B cells to express activation marker CD86. No significant differences were observed among cytokine production. Thus, the extra IFNα receptor may not help in B cell responses to infections and instead drive B cells into a more consistent inflammatory state.
Development and application of a high-content biology-based assay to quantify effects of modulators of NETosis in primary human neutrophils
Authors: Loomans C J M , Lin Xin-Xuan , Sciarrillo Rocco , Scandiuzzi Lisa , Stallen Jan , Stavenuiter Fabian , Vlaming Marijn 
Published: August 2023

Abstract
Neutrophils represent the first line of defense against pathogens by using multiple mechanisms, including phagocytosis, degranulation, and the release of neutrophil extracellular traps (NETs). NET formation can lead to a unique form of programmed cell death in neutrophils, called NETosis. In addition to its microbicidal function, compelling evidence has linked NETosis to the pathogenesis of diseases including autoimmune diseases, atherosclerosis and thrombosis.
To facilitate drug discovery programs for NETosis modulators, a high content biology-based method was developed and applied to blood derived primary human neutrophils. In summary, NETosis was triggered by PMA or BSA-Immune Complexes and the loss of membrane integrity together with spreading of NET formation beyond cell membranes as well as nuclear decondensation were captured in a live-cell setting on a CellVoyager (CV8000) platform. Furthermore, after addition of NETosis modulators (DPI and R406) to the triggered-neutrophils, a dose-dependent NETosis reduction was observed with optimal assay windows between 2.5 and 4 hours after trigger addition. Based on their distinct changes in nuclear morphology and loss of membrane integrity, this method was also able to distinguish various mechanisms of neutrophil cell death including NETosis and apoptosis. This procedure was successfully applied in medium-throughput screenings (96-well plate format), assessing dose-dependent inhibitory effects of selected large molecules, small molecules, and lipids on NETosis processes.

In conclusion
We developed a high content biology-based approach that can be employed to advance drug discovery for therapeutic areas associated with NETosis.
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GIMAP5 regulates a lymphocyte longevity assurance pathway
Authors: Gabrielski Justin Q , Park Ann Y , Lenardo Michael J 
Published: August 2023

Abstract
Elucidating the molecular mechanisms of immunodeficiency diseases is a powerful approach to discovering new immunoregulatory pathways in humans. One such previously unknown pathway is that of GIMAP5, an evolutionarily conserved GTPase expressed in T and NK lymphocytes, also known as the gene mutated in the Biobreeding diabetic rat and Sphinx lymphopenic mice. To better understand this disease association, we identified causative GIMAP5 mutations in 17 individuals of 9 kindreds with lymphopenia, liver disease, splenomegaly, thrombocytopenia, lymphadenopathy, lung infections, and lymphoma. As the mechanism of action, we now report that GIMAP5 controls the pathological accumulation of long chain ceramides. The ceramide synthase inhibitor, fumonisin B1, rescues GIMAP5-deficient T cells from ceramide overaccumulation and cell deterioration. Concordant with these findings, induction of ceramide accumulation and cell death by exogenous C6-ceramides in human T cell blasts was reduced by fumonisin B1 and GIMAP5 overexpression. Our work reveals the underlying long-chain ceramide mechanics of GIMAP5, opening new opportunities for further metabolomic studies of immunodeficiency diseases and targets for the development of therapeutics. This research was supported by NIAID by the Intramural Research Program of the NIH.
ITK inhibition improves the response to immune checkpoint blockade in solid tumors
Authors: Li Ling , Zhao Manzhi , Kiernan Caoimhe H , Castro Eiro Melisa D , Dammeijer Floris , van Meurs Marjan , Brouwers-Haspels Inge , Wilmsen Merel , Grashof Dwin G B , van de Werken Harmen J G , Hendriks Rudi W , Aerts Joachim G , Mueller Yvonne M , Katsikis Peter D 
Published: August 2023

Abstract
Cytotoxic CD8+ T cell (CTL) exhaustion is driven by persistent T cell receptor (TCR) stimulation in chronic infections and tumors. The exhausted state of T cells is characterized by impaired effector function and proliferative capacity, upregulation of multiple inhibitory receptors and altered transcriptional profiles. The phenotypic and epigenetic changes of these cells hinder effective antitumor immunity and immunotherapy. Therefore, reinvigorating exhausted T cells may provide significant clinical benefits to patients with cancer. Our approach is to reduce chronic TCR stimulation by inhibiting IL2-inducible T-cell kinase (ITK), a pivotal component of TCR signaling. In our study, we found that intermittent ITK inhibitor treatment significantly improved ICB response in three immune checkpoint blockade (ICB) -resistant solid tumors (melanoma, mesothelioma or pancreatic cancer) in vivo. In vitro, we demonstrated that specific ITK inhibitor and ibrutinib, an inhibitor of Bruton’s tyrosine kinase (BTK) and ITK, directly reversed the exhaustion-related phenotypes in exhausted CTLs. The changes included enhanced cytokine production, decreased inhibitory receptor expression and changes in transcription factors. Subsequently, RNA-seq data revealed that exhaustion-related transcriptional profile of in vitro CTLs is reduced by ibrutinib treatment. Ibrutinib mediated reversal of CTL exhaustion was BTK independent. To conclude, our study demonstrates that ITK inhibition can be used to directly ameliorate CTLs exhaustion and overcomes immune checkpoint blockade resistance in solid tumors.
This work was supported by the department of Immunology, KWF Grant 128371 (PDK), in part from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 779295 (PDK), and by the China Scholarship Council for funding PhD fellowships (No. 201506160120 for MZ and 201906210055 for LL).
A new PD-L1 antibody H1A causes PD-L1 degradation and sensitizes cancer cells for cytotoxic therapy
Authors: Tu Xinyi , Mutter Robert , Lou Zhenkun , Dong Haidong 
Published: August 2023

Abstract
PD-L1/PD-1 receptor pathway represents an important way of cancer cells to escape immune surveillance, yet only small portions of patients can benefit from PD-1/PD-L1 inhibitors. This is at least partially due to incomplete understanding of PD-1/PD-L1 biology in cancer. Although the interaction of extracellular domains between PD-L1 and PD-1 on cancer cells and immune cells is well studied, the function of PD-L1 inside cells, however, has not been well established. In our previous data, we found that intracellular PD-L1 acts as an RNA binding protein and interacts with NBS1 and BRCA1 mRNAs. Furthermore, intracellular PD-L1 protects targeted RNAs from degradation by RNA exosome, therefore increasing cellular resistance to DNA damage. Since Clinical approved PD-1/PD-L1 inhibitors have little effect on intracellular PD-L1, we sought to find new drugs to target intracellular PD-L1. Here we developed a new PD-L1 antibody, H1A, which has a different effect on PD-L1. Instead of blocking of PD-1/PD-L1 interaction, we found that this antibody disrupts the interaction of PD-L1 and CMTM6, which leads to degradation of PD-L1 by the lysosome. Therefore, H1A can unleash host antitumor activity and enhance the effects of DNA damaging therapy in cancer cells at the same time. Indeed, applying H1A in vivo significantly sensitized tumors to radiation and chemotherapy in a PDX models. Moreover, in a humanized immune competent TNBC PDX model, H1A monotherapy exhibited better therapeutic efficacy compared with clinically available PD-L1 inhibitor, suggesting that H1A activates anti-tumor immunity through PD-L1 degradation. Therefore, H1A may be a potential new drug to target both intracellular and extracellular PD-L1 to improve cancer therapy.
Neutrophils Extracellular Traps (NETs) Inhibition Improves PD-1 Blockade Immunotherapy in Triple-Negative Breast Cancer
Authors: Zhang Hongji , Fa Pengyan , Tsung Allan 
Published: August 2023

Abstract

Introduction
Despite significant advances using immune checkpoint blockade therapy for patients with advanced cancer, 40–85% of patients fail to respond to therapy. Increasing evidence suggests that the tumor immune microenvironment plays a critical role in rendering the immune checkpoint blockade ineffective. As such, targeting the tumor immune microenvironment is an attractive strategy for the treatment of solid tumors and an important approach to improve the efficacy of checkpoint inhibitors. Our study explores the combination therapy of DNase I and PD-1 antibody to prevent tumor growth. We demonstrate that this combination treatment for triple-negative breast (TNBC) cancer shows superior efficacy in comparison to a single-agent treatment, both in vivo and in vitro.

Methods
For 4T1 tumor models, 0.1-ml sample containing 104 tumors was injected into the mammary fat pad of female BALB/c mice. Tumor diameters were measured using calipers. The tumor volume was calculated. Anti-PD-1 or IgG2a isotype Ab (10mg/kg, every 3 days) and DNase I (5 mg/kg, daily) were administered intraperitoneally starting from day 5 after tumor cell inoculation.

Results
DNase I plays a pivotal role in the degradation of NETs, significantly dampening the resistance to anti-PD-1 blockade in a mouse TNBC model by attenuating tumor growth. Remarkably, DNase I decreases tumor-associated neutrophils and the formation of NETs in vivo. Mechanistically, the inhibition of NETs with DNase I results in the reversal of anti-PD-1 blockade resistance through increasing CD8+ T cell infiltration and cytotoxicity. These findings signify a novel approach to targeting the tumor microenvironment using DNase I alone or in combination with immune checkpoint inhibitors.

R01CA214865-06 (Renewal) Surgery triggered immune response and liver metastases. NIH/NCI 1R01 CA214865-01. Surgery triggered immune response and liver metastases.
Pharmacologic tumor PDL1 depletion as a novel approach to overcome treatment resistance
Authors: Murray Clare , Bai Haiyan , Ontiveros Carlos Oscar , Blinkiewicz Paige , Galvan Eva , Deng Yilun , Souto-Padron Alvaro , Curiel Tyler J 
Published: August 2023

Abstract
Tumor surface PDL1 canonically engages immune cell PD1 to inhibit anti-tumor immunity. However, PDL1 also mediates tumor-intrinsic pathologic signals, including promoting the DNA damage response (DDR) and suppressing immunogenic STING. Genetic tumor PDL1 depletion (CRISPR/Cas9, shRNA) induces novel treatment vulnerabilities that are not replicated by αPDL1 antibodies, but genetic tumor PDL1 depletion is not clinically feasible. We hypothesized that pharmacologic tumor PDL1 depletion could replicate genetic PDL1 depletion effects as a highly translational strategy to create novel treatment vulnerabilities. We conducted a high-throughput drug screen that identified 15 tumor PDL1 depleting drugs (PDDs) that potently reduce tumor PDL1 and are FDA-approved. Cefepime (β-lactam antibiotic) is a PDD that induced PDL1-dependent Chk2 DDR protein depletion (immunoblots), Chk1 inhibitor sensitivity in vitro (MTT, p&lt;0.0001) and in vivo in wild-type and NSG mice (p=0.03), and immunogenic STING signals (immunoblots, RT-qPCR). The PDD telmisartan (angiotensin II receptor blocker) induced PARP inhibitor sensitivity in vitro (p&lt;0.0001), impaired homologous recombination DDR (DR-GFP reporter, immunoblots), and induced STING signals and immunogenic cell death (flow cytometry). The PDD chlorambucil (alkylating agent) selectively depleted tumor PDL1 in vivo and sensitized αPDL1-resistant tumors to αPDL1 in a tumor PDL1 and NK cell-dependent manner. Thus, PDDs deplete tumor PDL1 to inhibit its pathologic cell-intrinsic signals, induce novel treatment vulnerabilities, improve tumor immunogenicity, and are rapidly translatable.
Supported by grants from NIH (Graduate Research in Immunology Program training grant T32 AI138944, TL1 TR002647, T32GM113896, CA204965, CA054515) and the Clayton Foundation (no grant number).
DRIVE: Diversity, Representation, and Inclusion for Value in Education – interprofessional faculty development for a bias-free curriculum at UMass Chan Medical School
Authors: Gerstein Rachel , Carter Yasmin , Haley Heather-Lyn , MacGinnis Christine , Peterson Kenneth , Hogan Susan , Whitehouse Amanda , Taweh Omar , Igiesuoboro Oghomwen , Herbert Carly , Fischer Melissa 
Published: August 2023

Abstract

Objective
The DRIVE Initiative engages interprofessional faculty and learners at UMass Chan Medical School to build tools and support for educators to identify and mitigate bias in their educational materials and teaching.

Methods
Key DRIVE resources include a core skills workshop, taught by representatives from the Graduate, Nursing and Medical schools. To encourage active application of these skills, there is a printable curriculum appraisal tool and an independent learning module. The DRIVE team also offers individual consultation for faculty as well as discussion across our academic community through weekly DRIVE-in cafés.

Results
In 2021, 16 core skills workshops were held with 84 faculty attending. Monday DRIVE-in cafés were started; the associated interactive library guide which links to curated resources and the anonymous reporting tool was accessed 922 times.

Discussion
While the workshop is required for core faculty in the School of Medicine and highly encouraged for all others, we have noted gaps in uptake particularly in basic sciences (Graduate School) and are increasing efforts in that area. Challenges have included demonstrating relevance to faculty across disciplines e.g., loose connection to basic science teaching. To increase adoption, materials and programs have been continuously updated and improved based on feedback, and workshops tailored to specific audiences. Recruiting compensated faculty from multiple schools has been vital to our success.

Conclusion
Biased curricula harm our learning environments and those who teach and learn within them, and lead to the perpetuation of disparities: the interprofessional approach of DRIVE has demonstrated success in mitigating this.
Assessment of factors that impact the quality and penetrance of immunology into the curriculum at the secondary education level in Alabama
Authors: Bruns Heather A , Evans Paulette , LaChenaye Jenna , Justement Louis B 
Published: August 2023

Abstract
Immunology as a discipline has significant relevance to health and disease and recent advances have harnessed the immune system to better fight against infectious diseases, as well as to address a wide range of illnesses that include allergy/asthma, autoimmunity, and the broad spectrum of inflammatory disorders. Therefore, immunology is a discipline that warrants increased attention not only at the graduate and professional school level, but at the undergraduate level. In spite of this, an analysis of approved undergraduate majors has shown that there are only a handful of programs at the undergraduate level that have a primary focus on understanding the immune system. Subsequent studies to understand the factors that influence the decision of individuals to pursue an undergraduate degree in immunology revealed that the primary means of exposure is through formal educational experiences and that the timing of the experience is critical. Studies demonstrated that exposure to immunology at a later stage, as an undergraduate versus in high school correlated with the decision to pursue immunology. It was hypothesized that if and when immunology is introduced at the high school level, either the students themselves are not prepared to master the material, or the educational experience is suboptimal. To further assess these possibilities, secondary educators were surveyed to determine the extent to which immunological concepts are integrated into the biology curriculum and the comfort level of the educators with immunological concepts. The study supports the need for programs to expand educator familiarity with immunology and to foster the development of learning activities to reinforce student learning and interest in immunology.
Department support only