bluuebunny/crossref_metadata_2025 huggingface dataset:
STINGing CD4+ T cells to induce refractoriness to HIV infection
Authors: Tomalka Jeffrey Alan , Sadek Naseem , Welbourn Sarah , Gordon David , Sekaly Rafick-Pierre
Published: August 2023

HIV rapidly infects CD4+ T cells where it can establish latent infection. Identifying immune mechanisms which can render CD4+ T cells refractory to infection represents a critical step in limiting infection and viral spread. Using purified memory CD4+ T cells from healthy donors, we found that activation of the STING signaling pathway with 0.5 mM of the STING agonist diABZI was sufficient to induce a state of refractoriness to in vitro HIV infection in memory CD4+ T cells, a major source of the HIV reservoir. We observed that STING activation in CD4+ T cells induces Type I IFN production and activates p-IRF3. CRISPR KD of STING in memory CD4+ T cells led to a significant increase in HIV infection, demonstrating a cell intrinsic role for STING signaling in limiting HIV infection/replication. Using proteomics, we identified that diABZI significantly and potently induces restriction factors and proteins involved in antiviral immunity and Type 1 IFN signaling. Importantly, proteins that were significantly reduced mapped to pathways including RNA processing and protein translation which are aspects of the host machinery which HIV needs to co-opt to replicate and produce infectious virions. Our data provide mechanistic insight into the STING pathway as a means to render CD4+ T cells refractory to HIV infection and highlights that this pathway lands a “1–2” punch by increasing expression of antiviral proteins that directly target the virus while suppressing the cellular machinery needed for the viral life cycle. The potential to use a STING agonist as a post-exposure therapeutic which directly limits viral infection while promoting global antiviral immunity represents a novel strategy that may stop HIV infection in its tracks.
CD127 and Sca1 mark self-driven memory-phenotype CD4+ T cells that differ from foreign antigen-specific memory cells and possess inflammatogenic and anti-tumor activities
Authors: Kawabe Takeshi , Ciucci Thomas , Kim Kwang Soon , Yang Ziying , Kawajiri Akihisa , Tayama Shunichi , Ishii Naoto , Jankovic Dragana , Zhu Jinfang , Sprent Jonathan , Bosselut Remy , Sher Alan
Published: August 2023

Under steady-state conditions, conventional CD4+ T lymphocytes are divided into naïve (CD44lo CD62Lhi) and memory (CD44hi CD62Llo) cell compartments. While the latter population is presumed to comprise a mixture of explicit foreign antigen (Ag)-specific “authentic” memory and foreign Ag-independent memory-phenotype (MP) cells, phenotypic markers differentially expressed in these two cell types have yet to be identified. Moreover, it is unknown whether MP cells consist of distinct subsets defined by marker expression. In this study we demonstrate using combined single cell RNA sequencing and flow cytometric approaches that self-driven MP CD4+ T lymphocytes are divided into CD127hi Sca1lo, CD127hi Sca1hi, CD127lo Sca1hi, and CD127lo Sca1lo subpopulations that are Bcl2lo while foreign Ag-specific memory cells are CD127hi Sca1hi Bcl2hi. We further show that amongst the four MP subsets, CD127hi Sca1hi lymphocytes represent the most quiescent, cell division-experienced subpopulation derived from peripheral naïve precursors. Finally, we provide new evidence arguing that the CD127hi Sca1hi MP subpopulation tonically expresses T-bet and possesses the capacity to trigger systemic inflammation including colitis, interstitial pneumonia, and nephritis as well as to exert anti-tumor activity against malignant lymphoma. Together our findings define MP CD4+ T lymphocytes as a unique, self-driven population consisting of distinct subsets that differ from conventional foreign Ag-specific memory cells in marker expression and reveal the inflammatogenic nature of the mature CD127hi Sca1hi MP cells.
HnRNPL is essential for T cell function in the Periphery
Authors: White Tristan L , Jin Ye , Morel Penelope A
Published: August 2023

While RNA binding proteins (RBPs) have recently emerged as regulators of gene expression in immune cells, their effect on T cell fate needs further investigation. Heterogeneous Nuclear Ribonucleoprotein (hnRNPs) are RBPs that play a crucial role in many aspects of RNA biogenesis, alternative splicing, expression, and function. During T cell development hnRNPL is known to regulate the proliferation, and migration of thymic pre-T cells, as loss of hnRNPL in early T cell development results in a failure of T cells to reach the periphery. Little is known about how hnRNPL affects T cell activation, and function. Our lab has used CD4 Cre × hnRNPLFl/Fl (KO) mice to understand the role of hnRNPL in peripheral T cells. Our initial study of the steady state profile of the KO mice shows normal migration of T cells form the thymus, but there is a reduction in peripheral T cell numbers. We also show that hnRNPL plays an important role in T cell activation and survival, since T cells lacking hnRNPL do not survive long enough to differentiate into T helper cell subsets. In vivo, using immunization studies, we show that KO CD4 T cells fail to fully differentiate into Tfh and are unable to support the formation of germinal center B cells. To ensure the effect that hnRNPL has on T cells in the periphery is not caused by the loss of hnRNPL during thymic T cell development we are using the CD4-Cre/ERT2 × hnRNPL (I-KO) to investigate how the loss of hnRNPL in peripheral T cell affects the ability of T cells to differentiate and survive post activation. Further studies will also focus on the mechanisms hnRNPL use to regulate T cells activation and survival. These studies suggest that hnRNPL plays an important role in the activation and survival of peripheral T cells.

AAI Fellowship
Single-cell Spatial Phenotyping of Immune and Metabolic Landscapes in Systemic Lupus Erythematosus
Authors: Jhaveri Niyati , Klymyshyn Dmytro , Ben Cheikh Bassem , Pratapa Aditya , Braubach Oliver
Published: August 2023

Autoimmune diseases like systemic lupus erythematosus (SLE) affect the health of multiple organ systems including the central nervous system, skin, joints, and kidneys. The heterogeneity of clinical profiles observed with SLE reflects a complex dysregulation of cellular processes, including metabolism, apoptotic clearance, and inflammation. Diagnosis and treatment of SLE is thus challenging, and few new treatments have been approved over the past 60 years. In this study, we aimed to characterize the spatial immune and metabolic profiles of SLE at single cell resolution. To do so, we profiled a cohort of human FFPE SLE tissues with an ultrahigh-plex panel of >50 oligo-conjugated antibodies representing immune, apoptotic, metabolic and stress markers. Whole-slide, spatial phenotyping was performed using the PhenoCycler®-Fusion system, followed by deep bioinformatic analyses to determine the cellular phenotypes, spatial neighborhoods, and metabolic determinants of SLE. By combining immune cell lineage information with expression of key proteins involved in glycolysis, amino acid metabolism, lipid metabolism, hypoxia, oxidative stress, and apoptosis, we elucidated the functional dysregulation in key immune subsystems that may contribute to development of autoimmunity and inflammation associated with SLE. Spatial phenotyping also revealed a high degree of inter- and intra-sample heterogeneity, confirming the complexity of the SLE clinical profile. Collectively, our data amount to new insights into metabolic reprogramming of immune cells in autoimmune diseases and may aid in the identification of novel therapeutic biomarkers within the tissue microenvironment of SLE patients.
The adaptation model of immunity: signal IV matters most in determining the functional outcomes of immune responses
Authors: Manjili Masoud H
Published: August 2023

Current research in immunology and immunotherapy is fully influenced by the self-nonself (SNS) model of immunity. This theoretical model suggests that alloreactivity results in graft rejection whereas tolerance towards self-antigens expressed by malignant cells facilitates cancer development. Similarly, breakage of immunological tolerance towards self-antigens results in autoimmune diseases. Accordingly, immune suppression is recommended for the management of autoimmune diseases, allergy, and organ transplantation while immune inducers are used for the treatment of cancers. Nevertheless, a cure for these human diseases remains elusive. This is in large part due to the application of the SNS model in the areas that are beyond its realm. The SNS model is designed to explain how an immune response is induced rather than to predict whether an immune response succeeds or fails against its target. Here, I propose the adaptation model of immunity to explain the decision-making process by activated T cells. According to this model, all somatic cells express adaptation receptors (AdRs) that relay survival signals upon engagement with their nominal adaptation ligands (AdLs) expressed on activated T cells. The AdR/AdL axis establishes signal IV to support immune-mediated tissue homeostasis. Any loss or alterations in the expression of tissue-specific AdRs results in immune intolerance or autoimmune diseases as well as graft rejection. Upregulation or downregulation of the AdRs by tumor cells could also dictate the outcomes of tumor-reactive immune responses. Therefore, novel therapeutics should focus on the discovery and targeting of the AdRs for the treatment of immune-related diseases and cancer.

This work was supported by the Office of the Assistant Secretary of Defense for Health Affairs through the Breast Cancer Research Program under Award No. W81XWH2210793. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the U.S. Department of Defense.. This work was also supported by multi-investigator pilot funding from the VCU Massey Cancer Centre, supported in part with funding from NIH/NCI Cancer Centre support grants P30 CA016059.
Type I Interferon-Dependent and -Independent TRIF Signaling are Required for Autoantibody Generation in an Induced Model of Systemic Lupus Erythematosus
Authors: Pilon Celeste , Subang Rebecca , Lonina Elena , Lesage Sylvie , Qureshi Salman T , Charette Emilie , Fonseca Gregory , Levine Jerrold S , Rauch Joyce
Published: August 2023

Systemic lupus erythematosus (SLE) is an autoimmune condition characterized by the production of autoantibodies to cellular and nuclear self-antigens, with damage to multiple organs. Immunization with the SLE autoantigen β2-glycoprotein I (β2GPI) and TLR4 ligand lipopolysaccharide (LPS) induces a murine model of SLE characterized by the generation of autoantibodies to multiple SLE antigens. LPS induces both inflammatory cytokines and type I interferons (IFNs) by signaling through the adaptor proteins MyD88 and TRIF, respectively. We investigated the mechanisms of LPS-dependent signaling on the induction of murine SLE following β2GPI/LPS immunization. Immunization of mice deficient in MyD88 or TRIF with β2GPI/LPS revealed that LPS-dependent TRIF signaling preferentially promoted autoantibody production in this model, compared to MyD88 signaling. Furthermore, SLE-specific autoantibody production was induced by TLR3 or TLR4 agonists, in combination with β2GPI, but not by agonists of TRIF-independent TLRs. RNA-sequencing of dendritic cells and macrophages isolated from wildtype (WT) and TRIF-deficient mice immunized with β2GPI/LPS revealed differential expression of both type I IFN-dependent and -independent genes. Moreover, type I IFN receptor (IFNaR)-deficient mice immunized with β2GPI/LPS showed diminished autoantibody production compared to WT mice, but to a lesser extent than TRIF-deficient mice. We conclude that LPS-dependent TRIF signaling is required for the generation of autoantibody production in our induced SLE model, both through type I IFN-dependent and - independent effects.

Supported by grants from CIHR (????)
Th17 cells of type 2 diabetic patients present an aberrant fatty acid metabolic pathway
Authors: Ujagar Naveena , Sriram Trupti , Chan Dennis , Proctor Elizabeth , Nicholas Dequina
Published: August 2023

Approximately 90% of the 537 million diabetic adults worldwide are living with type 2 diabetes (T2D). T2D is a metabolic disorder characterized by elevated blood glucose, which develops when a person is resistant to or cannot make enough insulin. Chronic systemic inflammation is a key contributing factor in the onset of insulin resistance and T2D. This inflammation in T2D patients stems from T-helper 17 (Th17) cells, likely due to defects in fatty acid metabolic pathways driving Th17 inflammation in T2D. While the importance of lipid metabolism has been shown in in vitro differentiated Th17 cells, the metabolism of endogenous Th17 cells in health or T2D has not been characterized due to their low abundance in vivo. We overcame this obstacle and established the relationship between defects in fatty acid metabolism and endogenous Th17 effector function with a novel Th17 cell enrichment assay. We captured CD4+ T cells with a hydrogel particle and magnetically purified them based on secretion of IL-17A/17F/21, allowing for us to perform scRNAseq on endogenous, activated Th17 cells. We then utilized an algorithm named Compass-powered by scRNAseq data and flux balance analysis, to characterize the cellular metabolic state of our captured Th17 cells. By overlaying scRNAseq data from endogenous Th17 cells of diabetic and control patients with gene ontology resources and applying in-silico modeling of metabolic pathways, we identified key lipid pathways driving Th17 inflammation in T2D and conducted the first study ever to be done on endogenous Th17 cells of healthy and T2D patients. This novel approach, and the data that ensued from it, with functional validation will illuminate inflammatory markers to target for creation of an immunotherapy for T2D.

Supported by grants from NIH (1DP2AI171121-01)
Ackr1-deficient mice are protected from lethal SARS-CoV-2 challenge
Authors: Majumdar Shamik , Pontejo Sergio M , Weaver Joseph , Murphy Philip M
Published: August 2023

To date, there have been more than 613 million confirmed cases of COVID-19, with over 6 million deaths worldwide. In severe cases, the causative agent of the COVID-19, SARS-CoV-2, induces acute respiratory distress syndrome. To study the roles of chemokine pathways during COVID-19 pathogenesis, we performed an infection screen in mice deficient in chemokine ligands and receptors using the mouse-adapted strain of the virus. Weight loss and death were recorded for 2 weeks after infection of wild type mice, and mice genetically deficient for Ccr2, Ccr5, Ccr6, Cxcr3, Cxcr6, Cxcl10 or the atypical chemokine receptor Ackr1. Acute weight loss was observed in all infected mouse strains; however, Ackr1−/− mice uniquely displayed markedly increased survival compared to wild type mice. ACKR1 (also known as the Duffy antigen receptor for chemokines) is a non-signaling receptor expressed on endothelial cells, erythrocytes and a subset of neurons that binds to a wide range of chemokines thereby controlling their availability as a scavenger and shaping chemotactic gradients. Comprehensive expression analysis revealed Ackr1 as the most highly induced chemokine receptor in infected lung post-infection. Of note, COVID-19 mortality is disproportionately low in West Africa, overlapping with the geographic distribution of genetic deficiency of erythrocyte ACKR1 due to a single nucleotide polymorphism in the ACKR1 promoter that interrupts binding of GATA-1. Further studies are required to understand the modulation of SARS-CoV-2 pathogenesis by Ackr1 and its potential to explain the trajectory and limited mortality of Covid-19 in Africa and to serve as a potential therapeutic target.
Host Gut-Resistome and Gut-Immune Axes are the key to Non-cholera Vibrio infection severity in aging and heat stress
Authors: Roy Subhajit , Saha Punnag , Bose Dipro , Chatterjee Saurabh
Published: August 2023

Global incidences of periodic heat waves and exposures are on the rise and that also have impacted non-cholera Vibrio abundance in coastal waters. We hypothesized that aging and a parallel heat stress (HS) may potentiate the risk of non-cholera Vibrio infections in the host and exacerbate the pathology. To study the underlying pathological changes in possible co-exposure of HS and subsequent non-cholera Vibrio challenge, an aging mouse model was used. Mice were exposed to HS at 40°C for three hours/day for 15 days. This was followed by a Vibrio vulnificus challenge 24 hours post HS exposure. Results showed that HS exposure decreased the diversity of the host gut microbiome and altered the relative abundance of many important taxa as assessed by Whole Meta-Genome Sequencing (WMS). Gut leaching was evident via the alteration of tight junction proteins and concomitant endotoxemia was evident. Decrease in abundance of Akkermansia muciniphila, a species well known for protection of gut barrier integrity was observed. Serum CRP levels, a clinical indicator of septicemia was elevated in the HS mice following Vibrio vulnificus infection, over and above the Vibrio-only or HS-only group. Significant amplification of gut leaching, gastrointestinal and systemic inflammation (IL-1β, IL-6, TNF-α), T-cell senescence and altering IgA levels were also noted in the HS+ Vibrio group. WMS also showed an altered host resistome signature and abundance of genes harboring the antimicrobial resistance to tetracycline group of antibiotics, a common antibiotic used for treatment of non-cholera Vibrio infections. In summary, HS augmented susceptibility to non-cholera vibriosis via host variability in resistome patterns and altered gut-immune mediators.

This study was supported by NIH/NIEHS Grant 1P01ES028942-01#Project 4 Toxicology awarded to Dr. S. Chatterjee.
Immunomodulatory functions of the gut protist Pentatrichomonas hominis in health and disease
Authors: Soto Ariadna Soledad , Engel Clarisse , Sanchez Luzmariel Medina , Mehta Isha , Meisel Marlies , Das Jishnu , Hinterleitner Reinhard
Published: August 2023

Protists have been identified as part of the mammalian gut microbiome and are known to impact gut immunity and complex intestinal diseases such as inflammatory bowel disease (IBD). In humans, studies mainly describe host interactions with disease-causing species, while little is known about how commensal protists impact the immune system. Protists from the Parabasalid family, including Pentatrichomonas hominis (P. hominis), have been identified in about 30% of healthy individuals; however, the role of these non-pathogenic commensal Parabasalids remains unstudied. Our data shows that P. hominis promotes Treg differentiation in vitro. Interestingly, this ability was found to be contact-independent, as culture supernatant was sufficient to induce Tregs. Moreover, by doing de-novo genome and transcriptome assembly and high-resolution mass spectrometry we have identified immunomodulatory candidate pathways and metabolites that might be mediating this effect. Re-establishment of tolerance is a major therapeutic strategy for treating intestinal diseases. Hence, we were interested in studying P. hominis impact on the gut immune landscape. To that aim, we successfully and stably colonized mice with P. hominis in gnotobiotic housing conditions and under antibiotic regimen and we are testing P. hominis role under steady state and IBD mouse models. Altogether, our research will address a present gap in knowledge, potentiating the study of protists in the microbiome and their association with intestinal disorders.
Fumarylacetoacetate hydrolase (FAH): a candidate host gene associated with susceptibility to tuberculosis
Authors: Reijneveld Josephine F , Nieto-Caballero Victor E , Innocenzi Gabriel , Ruvalcaba Angel , Budzik Jonathan , Gutierrez-Arcelus Maria , Suliman Sara
Published: August 2023

Tuberculosis (TB) disease remains the leading cause of mortality from an infectious disease globally, with an estimated 1.6 million deaths in 2021. TB is caused by infection with Mycobacterium tuberculosis (Mtb), resulting in variable outcomes from asymptomatic latent infection to clinical TB disease. However, most individuals exposed to Mtb never develop clinical disease. We hypothesize that host factors, including metabolic pathways, determine the outcome of Mtb infection. Based on prior data from human cohorts we observed an association between impaired tyrosine metabolism and progression to TB disease. We then reanalyzed a public dataset of Mtb-infected monocyte derived dendritic cells (DCs) and found that stimulation with either live or heat-inactivated Mtb resulted in downregulation of FAH (fumarylacetoacetate hydrolase), which encodes the last enzyme in the tyrosine catabolism pathway. Next, we used gene editing of FAH in conditionally immortalized myeloid progenitors (CIM) from mice to test whether downregulation of the FAH gene is associated with higher susceptibility to Mtb. We generated three independent fah-edited CIM lines, with two lines showing editing efficiency above 90%. We infected wildtype and fah-edited CIMs with an H37Rv-lux Mtb strain and detected significantly higher Mtb growth in fah-edited CIMs compared to wildtype CIMs (p=0.004). We also successfully optimized CRISPR in primary human monocyte derived DCs and macrophages to define the role of FAH during infection in physiologically relevant myeloid cell types. Our data suggest that FAH deletion compromises immune control of Mtb. We are currently exploring the immunoregulatory roles of FAH during Mtb infection, including inflammasome activation.
MANGO: Inferring gene regulatory networks from single cell multiomics
Authors: Williams Mark Elliott , Scharer Christopher D
Published: August 2023

Transcription factors (TFs) and their target gene regulatory networks (GRNs) control immune cell fate decisions during differentiation and responses to environmental cues and signals. To better understand how GRN control immune outcomes and are dysregulated in disease settings, we developed MANGO (Multiomics Aided Neural Graph Ontology), a computational method that integrates single cell gene expression and chromatin accessibility data to infer genome-wide, cell-type specific GRN. MANGO harnesses graph neural networks to generate representations of gene and regulatory link behavior and uses existing interpretable methods, like UMAPs, to visualize the GRN and gather insights about functional neighborhoods of regulators or programs of regulation. By scoring each regulatory interaction and aggregating those with common sources, TF activity can be quantified and compared between networks. We applied MANGO to a human PBMC dataset to define GRN and identify key regulatory, cell-type specific TFs, despite sometimes having low changes in expression. Additionally, we show that MANGO can distinguish cells by regulation network at single cell resolution by feeding each cell’s expression and accessibility through its respective cell type’s regulation network and projecting the resulting embedding in lower dimensions with a UMAP. Finally, MANGO was used to uncover memory B and T cell GRNs that prime rapid secondary immune responses. Given the increase in single cell multiome datasets, we anticipate MANGO will provide unique insights into GRN in normal and diseased immune responses, and ultimately give rise to patient specific understanding of the immune system and precision medicine.

Supported by grants from NIH/NIAID (R01 AI148471) to CDS
MHC Ib-restricted T Cells Generate the Dominant Memory T Cell Response in Early Stages of Mycobacterium tuberculosis Infection
Authors: Cui Yongyong , Shang Shaobin , He Ying , Wang Chyung-Ru
Published: August 2023

Unconventional T cells, which recognize monomorphic MHC Ib molecules, such as Qa1, Qa2, M3, CD1, and MR1 have been implicated in the primary response to Mycobacterium tuberculosis (Mtb) infection. However, whether these T cells play a role and which MHC Ib molecules participate in the memory response against Mtb infection remains unknown. To determine the optimal immunization strategy, we vaccinated mice that lack MHC Ia (Kb−/−Db−/−) with an attenuated Mtb H37Rv strain via intravenous (IV) or subcutaneous (SC) injection. We found both routes of immunization induced a comparable MHC Ib-restricted CD8+ T cell response after virulent Mtb H37Rv re-challenge. Since bacteria were cleared through the SC but not the IV route, we selected SC as our primary route of immunization. MHC Ib-restricted CD8+ T cells in vaccinated mice produced multiple proinflammatory cytokines, expanded faster after Mtb challenge than those in unvaccinated mice, and recognized several Mtb protein antigens. Importantly, the memory response of MHC Ib-restricted CD8+ T cells was more dominant than that of MHC Ia-restricted CD8+ T cells at early stages of infection. We also observed an increased percentage and number of Qa1, Qa2 and M3-restricted CD8+ T cells in vaccinated mice in the early stages of infection, suggesting that these MHC Ib molecules can elicit MHC Ib-restricted memory CD8+ T cell response. While CD1d-restricted NKT cells did not differ significantly between vaccinated and non-vaccinated mice, the activation kinetics of MR1-restricted T cells was accelerated in vaccinated mice. Overall, we show that MHC Ib-restricted T cells memory response can play an important role at the early stages of Mtb infection thus should be included in subunit vaccine formulation.

Supported by grants from NIH (R01AI141083)
Subsecond release of catecholamines from CD4+ T lymphocytes
Authors: Seibold Jordan M , Ross Ashley
Published: August 2023

The neuroimmune system is complex and is regulated by small chemical messengers released from resident immune and neuronal cells. Catecholamines like dopamine and norepinephrine are important chemical messengers responsible for initiating and propagating messages between neurons and immune cells; however, the mechanism and dynamics of this signaling is relatively unknown. The presence of the intracellular machinery to synthesize and interact with catecholamines in T lymphocytes has been known for over a decade; however, direct measurement of catecholamine secretion in real time from immune cells has not been possible. Here, we have discovered that CD4+ T cells are capable of releasing catecholamines on a subsecond time scale. We have used a revolutionary technique, often used in the neuroscience field, called fast-scan cyclic voltammetry (FSCV) to measure subsecond fluctuations of catecholamines from isolated CD4+ T cells. This work provides direct evidence that T cells are capable of releasing catecholamines; potentially as a feedback communicator to resident sympathetic neurons in host immune organs. These results also prove that immune cells can signal on the same time scale as neurons . This work will hopefully open the door to enable more sophisticated studies at the neuron-immune synapse in the future.

National Institute of Health R01AI151552
Pre-activation of follicular B cells in old mice generates a hyper-response during germinal center reaction
Authors: Ticas Rodas Carlos J , Maul Robert W , Gearhart Patricia J
Published: August 2023

Immune cells acquire a variety of defects that accumulate overtime in a process known as Immunosenescence. To study any innate defects that B lymphocytes may acquire during aging, we have developed an adoptive transfer system using follicular B cells from young (FOY, 8–12 weeks) and old (FOO, >22 mo.) mice into a μMT B cell-deficient mouse. This model reveals that FOO cells, compared to their young counterparts, produce a higher amount of activated germinal center (GC) B cells in the presence of the common antigen NP-CGG after 14 days of immunization. However, these cells do not produce antigen-specific antibodies while showing a significant decrease in NP+ λ+ antigen-specificity. To study this change in phenotype further, we performed single cell RNA sequencing using both young and old populations of naïve, and day 14 GC B cells. Comparison of the naïve populations by PCA showed only minor differences between populations. Interestingly, Ccr6 is being upregulated in FOO cells but has a decreased expression in the old population of GCs. This change in expression suggests that FOO cells are pre-activated before transfer to μMT mice and antigen encounter. Analysis of the immunoglobulin repertoire in these cells showed that FOO cells also have decreased use of the canonical NP-specific IgH V-gene 1–72 and fail to include the CDR2 W33L mutation that is known to increase antibody affinity 10-fold. Taken together, these findings show that old mice produce pre-activated and hyper-responsive cells that cause a lack of proper selection and affinity maturation during the GC reaction.

This research was supported in part by the Intramural Research Program of the NIH, National Institute on Aging.
Spatial Profiling of Glioblastoma Microenvironments and Response to Immunotherapy
Authors: Garcia-Rivera Lizmarie , Chen Dan , Kaech Susan
Published: August 2023

Glioblastoma (GBM) is the most common adult brain cancer. Recent therapeutic advances, including immunotherapies that enhance anti-tumor immune responses, have failed to significantly improve survival. To address these failures, deeper studies on the cellular interactions that orchestrate whether GBM resists or regresses during immunotherapy are needed. Our group recently discovered that anti-CTLA-4 therapy initiated a novel microglia-CD4 T cell partnership. Following anti-CTLA-4, tumor-bearing mice showed a significant increase in survival and enhanced tumor regression. Further investigation showed that CD4 T cells, not cytotoxic CD8 T cells, were necessary for this response and that microglia were responsible for both sustaining tumoricidal CD4 T cell activity via MHC-II antigen presentation and phagocytosing debilitated tumor cells. With the rise of spatial profiling platforms, such as NanoString’s CosMx, we can now capture snapshots of the cellular dynamics that lead to a successfully mounted anti-tumor response in a continually evolving tumor microenvironment. Using both murine models and human GBM specimens, we are exploring the cellular networks that support beneficial CD4 T cell-microglia interactions and tumor regression. Conversely, we are also examining the role of regulatory T cells in the periphery and tumor microenvironment, and how these cells may interact with other immune suppressive niches, including GBM-associated microglia and infiltrating macrophages, to support GBM progression. Through this work, we are mapping the GBM cellular niches that support or suppress anti-tumor responses, with the goal of exposing vulnerabilities that can be exploited with better tailored immunotherapies.

Supported by grants from NIH (R01 CA240909, R01 CA230275)
Determining mechanisms of T cell leukemogenesis using E2A−/− mice
Authors: Parriott Geoffrey R , Bartom Elizabeth T , Kee Barbara L
Published: August 2023

T cell acute lymphoblastic leukemia (T-ALL) is a hematological malignancy caused by the transformation of T-lymphocytes during their development. 60% of pediatric T-ALLs have ectopic expression of Tal1, a gene that promotes leukemogenesis in part by altering the function of E2A. Consistent with this, mice with germline deletion of E2A develop T-ALL that mirrors human Tal1+ T-ALL. In this study, we used mice that delete E2A at different stages in T cell development to investigate the mechanisms causing T-ALL. Mice carrying a VavCre transgene and floxed alleles of E2A (VcKO) delete E2A early in T cell development, have reduced numbers of thymocytes prior to transformation, and develop T-ALL with an average latency of 18 weeks. Mice with an LckCre transgene (LcKO) delete E2A later than VcKO mice, have replete thymi, and rarely develop T-ALL. From this, we hypothesized that 1) E2A regulates a gene program in early T cell development that is antagonistic to transformation, or 2) Having a replete, competitive thymus suppresses transformation. Pre-leukemic VcKO thymocytes showed significantly more differentially expressed genes than LcKO thymocytes, including Notch1 and Myc, and are enriched for cancer associated pathways. To determine if the lack of competition in the VcKO thymus contributes to leukemogenesis, we reconstituted VcKO thymi by transplanting wild type bone marrow into sublethally irradiated VcKO mice. BM transplant significantly increased the number and frequency of DP thymocytes and increased leukemia latency. Together, these data suggest that intrinsic loss of E2A promotes leukemogenesis by changing the gene program of early developing thymocytes and by creating a non-competitive environment that supports transformation.
Robust eIF4B levels undermine invasive growth and immune evasion mechanisms in murine triple negative breast cancer models
Authors: Barbi Joseph J , Smith Randall , Diba Leila Zabihi , Srinivasan Aravind , Ossevoort Thomas , Zollo Robert , He Qian , Venkatesh Meera , Arora Aayush P , Washington Deschana , Yao Song , Walker Sarah E
Published: August 2023

Dysregulated protein synthesis is seen in many aggressive cancers, including metastatic breast cancer. However, the specific contributions of certain translation initiation factors to in vivo disease remain undefined. This is particularly true of eIF4B, an RNA-binding protein and cofactor of the RNA helicase eIF4A and associated eIF4F cap-binding complex. While eIF4A, eIF4G, and eIF4E are known to contribute to the progression of many cancer types, the role played by eIF4B in breast cancer remains relatively unclear. We therefore explored how naturally divergent and experimentally modulated eIF4B levels impact tumor growth and progression in murine triple negative breast cancer (TNBC) models. Surprisingly, we found that higher eIF4B levels in mouse and human breast cancers were associated with less aggressive phenotypes. shRNA-mediated eIF4B knockdown in TNBC lines failed to markedly alter proliferation and global translation in cells grown in vitro and only modestly hindered primary mammary tumor growth in mice. However, eIF4B knockdown significantly enhanced invasive growth in vitro and exacerbated both tumor burden and mortality relative to nontargeting shRNA controls in a model of metastatic disease. Deposited patient data reinforced a link between high eIF4B levels and better outcomes. Interestingly, in vitro and in vivo studies associated low eIF4B expression with more formidable immune evasion. These findings suggest that eIF4B levels impact metastatic disease in part through immunomodulatory mechanisms beyond the factor’s traditional role supporting eIF4/F directed translation. Future studies should explore the impact of these findings for personalizing treatment decisions and developing future therapies.
Targeting the NKG2A axis with a TCR mimic antibody containing an active Fc domain promotes anti-tumor immunity
Authors: Ghaffari Soroush , Weidanz Jon
Published: August 2023

The NKG2A axis is an emerging immune checkpoint. Blocking antibodies to the NKG2A receptor in combination with cancer vaccines and PD-(L)1 antibodies have been found to enhance anti-tumor immunity in murine models and cancer patients. The ligand for the NKG2A receptor is the non-classical major histocompatibility (MHC) class Ib molecule, Qa-1b in mice (HLA-E in humans), loaded with leader peptides from classical MHC class Ia molecules to form Qa-1b/Qdm peptide complexes. Here, we explore the therapeutic value of targeting the NKG2A ligand, not done previously, using our recently described single-domain T-cell receptor mimic (TCRm) antibody, EXX-1. Two syngeneic murine tumor models (A20 and CT26) were used to evaluate the therapeutic potential of EXX-1 and its mode(s) of action. The initial study done showed EXX-1 Fc active (mIgG2a) displayed optimal tumor growth inhibition and prolonged survival time in mice compared to a modified form of EXX-1 containing an inactive Fc domain. These findings suggested that in addition to blocking the NKG2A pathway, EXX-1 Fc active antibody likely induces tumor cell death via antibody dependent cellular cytotoxicity (ADCC). Moreover, mice treated with EXX-1 antibody as single agent therapy showed significantly better survival outcomes compared to the cohort of mice treated with blocking antibody to the NKG2A receptor. This implies that there might be potential benefits of targeting the NKG2a ligand over the NKG2A receptor though this will require further investigation. Additionally, combination therapy using EXX-1 Fc active and PD-(L)1 blocking antibodies revealed markedly improved rates of tumor-free mice compared to mice receiving treatment with EXX-1 Fc active or anti-PD-(L)1 antibody alone.
Relapsing subcutaneous nodules of the penis in a pediatric patient
Authors: Sechi Andrea , D'Amore Emanuele Stefano Giovanni , Naldi Luigi , Bellettato Massimo , May Lee Marco , Cutrone Mario
Published: August 2023

A 14-year-old boy presented with a history of non-tender, subcutaneous coalescing nodules located on the ventral-lateral aspects of the penis shaft for one year. Laboratory investigations for blood count and autoimmunity were within normal limits. Complete excision was performed, and on histology, the dermis showed necrobiotic material composed of altered collagen bundles, surrounded by a palisade of histiocytes and scattered lymphocytes, thus allowing a diagnosis of subcutaneous granuloma annulare. Only 18 published cases reported penile granuloma annulare. Medical management was advocated in 7/18 cases, either as a first-line or adjuvant therapy where surgery was not radical. Three patients received high-potency local steroids: two cases underwent adjuvant sessions of intralesional triamcinolone, and one patient received pentoxifylline orally. Surgery should be considered a second-line option since 5/8 of treated cases eventually recurred. The pentoxifylline-treated case witnessed a relapse after drug discontinuation, while topical steroids lead to complete recovery without relapses.
Association of IL4RA and IL-8 polymorphism in predicting susceptibility towards tuberculosis
Authors: K Jyotish , Joshi Jawahar Lal , Kansal Heena , Chopra Vishal , Garg Kranti , Girdhar Nidhi , Sharma Karan , Deshwal Rishabh , Sharma Siddharth
Published: August 2023

Mycobacterium tuberculosis is an infectious bacterial disease frequently affecting the lungs. With two fatalities from tuberculosis (TB) occurring every three minutes, India has the highest disease burden. The aetiology of tuberculosis has been linked to IL-8 and IL-4RA. Thus, the impact of the IL4RAQ576R and IL8 gene polymorphism on TB susceptibility was assessed. 301 healthy and 301 TB patients participated in a cross-sectional study. PCR RFLP was performed to identify the genotype of the IL4RAQ576R and IL-8 +781C/T gene polymorphism. The odds ratio and 95% confidence intervals were calculated using logistic regression to evaluate the risk of TB with IL4RAQ576R and IL-8 +781C/T polymorphism. A significant association was found between IL-4RA (p=0.04) and IL-8 +781 C/T (p= 0.03) in tuberculosis. Further, when clinical symptoms were compared with both polymorphisms, two of them, i.e., cough in IL-4RA576R (p=0.04) and breathlessness (p=0.01) in IL-8 +781C/T, showed a significant association. Moreover, different combinations of the SNPS were made, and the 3 risk allele shows a significant protective role (p=0.02). There is considerable evidence which shows that M. tuberculosis causes TB, an infectious disease that is genetically predisposed. The results of our study also showed that IL-4 RA Q576R and IL-8 +781 C/T played a significant protective function against tuberculosis, confirming the claim mentioned earlier. However, only the cough in IL-4RA576R and the dyspnea in IL-8 +781C/T exhibited a significant co-relation in TB patients when symptoms were examined. Additionally, the combined effects of the two SNPs were investigated, and it was discovered that the 3-risk allele has a strong association with tuberculosis. Therefore, the polymorphisms mentioned earlier, which may also be influenced by ethnicity, may significantly impact the chance of developing tuberculosis.
Analyse of Febrile Neutropenia Attacks in Children with Acute Lymphoblastic Leukemia
Authors: Karadoğan Meriban , Mutlu Fatma Türkan
Published: August 2023

Febrile neutropenia (FN) is the most important cause of morbidity and mortality during treatment in childhood with acute lymphoblastic leukemia (ALL). The aim of this retrospective study was to assess clinical features, outcomes, treatment modalities, documented infection rates, and frequency of isolation of specific organisms from ALL patients treated by our Department of Pediatric Hematology and Oncology using Berlin-Frankfurt-Munich (BFM) protocols. In this study 132 children diagnosed with ALL and treated according to the ALL-BFM 2009 protocol at Kayseri City Hospital, Department of Pediatric Hematology and Oncology between July 2015 and May 2022 were enrolled and evaluated for episodes of FN during intensive chemotherapy. Treatment characteristics, the presence of relapse, duration of neutropenia, culture results, the choice and duration of antibiotics, and disease prognosis were retrospectively assessed using patient records. In 132 acute leukemia cases, 278 episodes of FN were observed aged 1.1 and 17.8 years (mean 7.1±4.9 years) in a male to female ratio of 1.5:1. Infection focus could be documented in 66 episodes (23.7%); pulmonary infections in 23 patients (34.8%), gastrointestinal tract infections in 20 patients (30.3%), in 10 patients urinary tract infections (15.1%), derma¬otological and soft tissue infections in 2 patients (3%). The causative infectious agent could be demonstrated in 92 (33%) episodes. The most common site of isolation was blood (86/92, 30.9%). While the most frequently identified bacterial infectious agents were Gram-positive bacteria (56 isolates, 20.1%), Gram-negative bacteria were identified in 28 isolates (n=10%). Fungal growth was detected in 6 (2.1%) patients and polymicrobial growth was detected in 3 (1.1%) patients. Of the 132 patients, 4 (3%) died due to infection 2 died during induction, and 2 died during reinduction phases. Centers should evaluate their results to identify changing epidemiological patterns and to treat FN early and efficiently.
Arboreal Categories: An Axiomatic Theory of Resources
Authors: Abramsky Samson , Reggio Luca
Published: August 2023

Game comonads provide a categorical syntax-free approach to finite model
theory, and their Eilenberg-Moore coalgebras typically encode important
combinatorial parameters of structures. In this paper, we develop a framework
whereby the essential properties of these categories of coalgebras are captured
in a purely axiomatic fashion. To this end, we introduce arboreal categories,
which have an intrinsic process structure, allowing dynamic notions such as
bisimulation and back-and-forth games, and resource notions such as number of
rounds of a game, to be defined. These are related to extensional or "static"
structures via arboreal covers, which are resource-indexed comonadic
adjunctions. These ideas are developed in a general, axiomatic setting, and
applied to relational structures, where the comonadic constructions for
pebbling, Ehrenfeucht-Fra\"iss\'e and modal bisimulation games recently
introduced by Abramsky et al. are recovered, showing that many of the
fundamental notions of finite model theory and descriptive complexity arise
from instances of arboreal covers.
Klasifikasi Kematangan Buah Apel Berdasarkan Warna Dan Tekstur Menggunakan Algoritma K-Nearest Neighbor
Authors: Indra Pratama Fandy , Wijaya Akhmad Pandu , Pratiwi Hasti , Budianita Avira
Published: August 2023

Apel (Malus Domestica) adalah salah satu jenis buah yang unggul dan sangat digemari dan dikonsumsi masyarakat. Buah apel banyak digemari masyarakat karna rasanya yang bervariasi. Buah apel manalagi yang masih belum matang memiliki warna hijau dan untuk apel manalagi yang sudah matang memiliki warna hijau kekuningan. Hal tersebut tentu akan sedikit menyulitkan untuk membedakan antara buah apel manalagi yang belum matang maupun yang sudah matang apabila tidak terlalu memiliki ilmu mengenai dunia pertanian terutama pada buah apel. Sebab orang awam dirasa akan sulit membedakan. Sehingga akan diperoleh penilaian yang berbeda oleh setiap individu yang menyebabkan tingkat akurasi yang berbeda. Oleh karena itu dibutuhkan suatu teknologi yang dapat menentukan nilai tingkat akurasi dari pengklasifikasian kematangan buah apel manalagi sehingga memperoleh nilai yang konsisten. Penelitian ini bertujuan untuk menghasilkan prototype untuk mengklasifikasikan buah apel menggunakan Metode K-Nearest Neighbor berdasarkan tingkat kematangan yang direpresentasikan dari sisi warna dan teksturnya dan membantu memudahkan masyarakat dalam menentukan jenis buah yang matang, sedang, dan mentah. Diharapkan adanya teknologi ini dapat mempermudah manusia dalam mengklasifikasikan tingkat kematangan buah apel secara akurat. Metode pengujian ini menggunakan 12 buah apel yang terdiri dari matang, sedang, dan mentah yang menghasilkan tingkat kematangan buah apel sebesar 91,6667% dari total akurasi
Analisis Dan Visualisasi Data Body Performance Menggunakan Tiga Tools Visualisasi
Authors: Husna Lailatul , Prasetyo Utomo Pradita Eko
Published: August 2023

Artikel ini membahas tentang Analisa dan visualitasasi data dari data body performance .dimana nanntinya hasil Analisa penulis akan dicantumkan dibagian hasil dan pembahasan.dari artikel ini kita dapat melihat keuntungan memvisualisasikan data yang ada.dengan adanya beragam tools visualisasi data akan mempermudah kita dalam menganalisis dan memvisualisasikan data yang diinginkan.berikut tiga tools yang digunakan dalam artikel ini yaitu : Microsoft Power BI, Google Colab, dan Tableau Public.Adanya visualisasi data akan mempermudah mengambil keputusan.Dalam melakukan visualisasi data harus memperhatikan beberapa hal seperti data apa yang akan digunakan , apa yang ingin divisualisasikan , dan dugaan apa yang ingin dibuktikan.

Kata kunci—analisis dan visualiasi data, Tableau Public, Microsoft Power Bi, Google Colab.
The Role of Narcissism and Attachment in Adult Romantic Relationships: A Study of Greek-speaking Adult Participants
Authors: Ellina Eleni , Parpottas Panagiotis
Published: August 2023

The present study examined the relationship between narcissism and attachment dimensions in adult romantic relationships. One hundred and fifty one adults from the general population completed self-report measures of attachment dimensions, grandiose and vulnerable narcissism. The results indicated that both types of narcissism were significantly correlated to insecure attachment dimensions (anxiety and avoidance). Additionally, the results of the hierarchical multiple regression initially showed a significant prediction of grandiose narcissism in the insecure attachment dimensions, which was then counterweighted by the stronger contribution of vulnerable narcissism. The most significant variable in the overall examination of narcissism was vulnerable narcissism, significantly predicting both insecure attachment dimensions. Results are discussed in light of attachment theory and implications for clinical practice and future research are also considered.