bluuebunny/crossref_metadata_2025_split huggingface dataset:
Eos regulates IL-7 signaling and central memory T cell differentiation
Authors: Pokhrel Srijana , Read Kaitlin A , Jones Devin M , Powell Michael D , Warren Robert T , Oestreich Kenneth J 
Published: January 2023

Abstract
CD4+ central memory T cells (TCM) are key players in recall immune responses and thus are critical to long-term immunity. Despite their importance, much remains to be defined regarding the mechanisms that promote their differentiation and survival. Ikaros zinc finger (IkZF) transcription factors are known regulators of T and B cell differentiation programs. To date, the IkZF factor Eos has been mainly linked to regulatory T cell suppressive functions. Now, we have found that Eos is expressed at elevated levels in CD4+ TCM populations. Comparison between WT and Eos-deficient T cells in a murine model of influenza infection revealed a decrease in the percentage of CD4+ TCM cells in the absence of Eos, suggesting that Eos promotes TCM differentiation and/or survival. Using an established in vitro model of CD4+ TCM-like cell differentiation, we found that loss of Eos correlated with reduced expression of genes encoding key TCM transcription factors and cell surface markers. Further, Eos deficiency resulted in decreased STAT5 activation downstream of IL-7 signaling, a known positive regulator of TCM populations. In WT cells, IL-7 treatment increased Eos expression, suggesting a positive feed-forward relationship exists between Eos expression and IL-7/STAT5 signaling. Overall, our findings demonstrate that Eos is an important regulator of TCM populations. Understanding the mechanisms by which Eos regulates TCM function and survival may therefore be of interest in exploiting TCM protection against re-infection for therapeutic benefit.
Supported by a grant from NIAID (NIH- R01 AI134972) and funds through The Ohio State University College of Medicine
The strength of antigenic and IL-2 signals epigenetically programs functional memory CD8+ T cells
Authors: Guillen Erik , Chin Shu Shien , Chorro Laurent , Achar Sooraj , Ng Karina , Oberle Susanen , Alfei Francesca , Zehn Dietmar , Altan-Bonnet Gregoire , Delahaye Fabien , Lauvau Gregoire 
Published: January 2023

Abstract
Priming of naïve CD8+ T cells requires three signals: 1) cognate antigen, 2) co-stimulation, and 3) cytokines. These signals orchestrate CD8+ T cell differentiation. Particularly, strong T cell receptor (TCR) signals induce a more robust effector response, while weak TCR signals preferentially favor memory CD8+ T cells. However, memory cells induced upon varied TCR signaling strengths have been reported to be functionally similar. In this work, we show that, in contrast to this assumption, the strength of T cell receptor (TCR) signaling determines the requirement for interleukin-2 (IL-2) signals to form a pool of memory CD8+ T cells that are functionally different. We found that upon secondary antigen encounter, memory CD8+ T cells induced upon priming with lower TCR and IL-2 signals exhibited delayed cell cycle entry, impaired expansion and intracellular calcium accumulation compared to those primed with strong TCR and IL-2 signals. Furthermore, we also establish that the combination of both robust TCR and IL-2 signals, but not either one alone, are necessary to induce genome-wide changes in chromatin accessibility in regions targeting a wide range of biological processes consistent with their greater functional fitness. These notably included greater chromatin accessibility in promoters of genes encoding for stem cell, cell cycle and calcium-related proteins. Taken together, these findings demonstrate that modulating TCR strength and IL-2 signaling at the time of CD8+ T cell priming, lead to the differentiation of functionally distinct pools of memory CD8+ T cells.
Supported by T32GM7288
IL-23 signaling limits ferroptosis-driven immunopathology during systemic fungal infection
Authors: Millet Nicolas Olive Jean , Solis Norma V , Aguilar Diane , Lionakis Michail S , Wheeler Robert T , Jendzjowsky Nicholas , Swidergall Marc 
Published: January 2023

Abstract
Pattern recognition receptors initiate antifungal immune responses during infection. Recently, we identified that the ephrin type-A receptor 2 (EphA2) is a fungal β-glucan receptor which orchestrates innate immune responses and prevents oral Candida infection. However, the role of EphA2 during invasive candidiasis (IC) remains unknown. Here we found that EphA2 induces renal inflammation and injury; thus, promoting immunopathology during IC. In Epha2−/− mice, dendritic cells migrated more efficient in infected tissues, while secreting more IL-23. Furthermore, Epha2−/− mice had reduced lipid peroxidation in kidneys, a marker of ferroptotic cell death. Pharmacological inhibtion of ferroptosis increased macrophage fungal killing, reduced inflammation, and increased disease outcomes during IC. Previously, IL-23 signaling has been associated with myeloid cell survival during candidiasis. Treatment of macrophages with IL-23 reduced ferroptotic macrophage cell death and excessive inflammation, while increasing killing capacity. Furthermore, IL-23 treatment of infected mice increased survival and reduced ferroptosis during IC. Collectively, our study demonstrates that ferroptotic host cell death is linked to immunopathology and can be targeted by recombinant cytokine therapy during fungal infection.
This work was supported in part by NIH grant R00DE026856, an American Association of Immunologists Careers in Immunology Fellowship, and a TLI Seed grant to MS
Developing a transgenic mouse model to better understand the role of CD8 and granulysin against Mycobacterium tuberculosis infection.
Authors: Thakur Preeti , Sutiwisesak Rujapak , Lu Yu-Jung , Behar Samuel 
Published: January 2023

Abstract
Human cytotoxic lymphocytes kill infected cells through release of lytic proteins contained in cytotoxic granules. The cytotoxic granules of human NK and CD8 T cells contain perforin, granzyme and granulysin (GNLY), and GNLY has direct antimicrobial activity against Mycobacterium tuberculosis (Mtb). Murine studies suggest that CD8 T cells have only a modest role in immunity to tuberculosis (TB). We hypothesize that mouse CD8 T cells are ineffective at containing Mycobacterium tuberculosis (Mtb) as they don’t express GNLY. To test this hypothesis, GNLY-transgenic mice that express human granulysin protein under the control of human regulatory elements were infected with Mtb. GNLY-Tg and non-tg control mice control Mtb similarly. There was no difference in the ability of GNY-Tg and non-tg CD8 T cells to transfer protection to T cell deficient mice. Anti-CD3 stimulation led to expression of granulysin in human CD8 T cells, but not in murine CD8 T cells. However, after Mtb infection, GNLY was expressed in NK cells but not CD8 T cells. We conclude that GNLY-expressing NK cells do not alter resistance to Mtb. Importantly, GNLY-Tg mice are inadequate model to study CD8 function during TB. We are developing a new strategy to express granulysin in CD8 T cells. We expressed GNLY (isoform 2) in primary murine CD8 T cells by retroviral gene transfer and detected GNLY expression in 60% of CD8 T cells.

Future directions
We are assessing the cytolytic activity of GNLY-expressing CD8 T cells and their ability to kill intracellular pathogens in vitro and in vivo. We expect to develop a new GNLY-Tg model that can be used to study the function of CD8 T cells and granulysin in many different diseases. This work is supported by NIH AI159374.
This work is supported by NIH grant R21 AI159374.
Sphingolipids: A double-edged sword in the defense against Mycobacterium tuberculosis.
Authors: Guzman Gaelen D , Leier Hans , Niekamp Patrick , Holthuis Joost , Tafesse Fikadu G 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb) is a pathogen with a massive global health impact, and yet fundamental questions remain regarding the underlying mechanisms of its entry and subversion of innate immune mechanisms – particularly the manners in which host lipids influence the course of an infection. This pathogen gains entry to host alveolar macrophages via phagocytosis and, within the phagosome, Mtb perturbs host lipid metabolism and distribution. Here, we’ve shown that Mtb relies on an intact sphingolipid biosynthesis pathway to gain entry to host cells but that sphingolipids restrict the intracellular growth of this pathogen. We used genetic ablation and small molecule inhibition to perturb sphingolipid biosynthesis, resulting in reduced Mtb uptake across multiple phagocytic cell types without affecting receptor mediated endocytosis. Mechanistically, we visualized the dynamics of several signaling events during phagocytosis to demonstrate that sphingolipid deficiency results in poor initiation of phagocytic signaling. Intriguingly, sphingolipid inhibition at later stages of infection may result in enhanced intracellular Mtb growth and accelerated lysosomal damage and subsequent phagolysosomal escape. This study has revealed that the sphingolipid family may constitute a double-edged sword which plays multiple roles during Mtb infection. In understanding the interplay between Mycobacterium tuberculosis and host factors such as sphingolipids, we hope to identify novel methods of treatment for this ancient and global pathogen.
Supported by grants from NIH (R21 3R21AI124225-01A1 and R01 1R01AI14154 9-01A1)
A GABA-producing probiotic for the protection of CNS demyelinating inflammation
Authors: Ochoa-Reparaz Javier , Hoffman Kristina , Long Tyrel , Doyle William J , Kohl Hannah M , Staben Kendall , Sargent Alivia , Linton Rachel , Ristig Molly , Harris Rylee , Shi Xutong , Gibson K Michael , Roullet Jean-Baptiste , Castillo Andrea R 
Published: January 2023

Abstract
The gut-microbiota-brain axis has emerged as a critical pathway in the regulation of neuroinflammation. The gut microbiome regulates the severity of many experimental models of autoimmune central nervous system (CNS) demyelinating inflammatory diseases. Our most recent findings demonstrate that the microbiota of mice from different sources affects the severity of CNS inflammatory demyelination. Neuroinflammation triggered in experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis, modified the gut microbiota composition. The disease progression resulted in a significant reduction in the relative abundances of members of lactic acid bacteria when compared to healthy control mice. Among the altered taxa, bacteria producing gamma-aminobutyric acid (GABA) were significantly reduced. We hypothesized that modifying the microbiota with a probiotic while increasing intestinal GABA levels would reduce EAE’s severity. We genetically engineered a Lactococcus lactis probiotic that produces excessed levels of GABA. Real-time quantitative PCR analysis demonstrated an elevated expression of glutamic acid decarboxylase (GAD). GABA-specific ELISA showed a significant increase in neurotransmitter production when exposed to increasing concentrations of glutamic acid and time. In vivo, five times/week oral gavages with 5 × 108 CFU/mouse of GAD L. lactis but not with empty-plasmid carrier L. lactis protects against EAE in mice compared with sham-treated mice and prevents weight loss of animals while modulating the microbiome’s composition. Our results show that the increase of GABA at the intestinal level with the oral treatment with a probiotic strain protects against neuroinflammation in the CNS.
Supported by grant from NIH (R15 NS107743)
Aging impairs regulatory T cell (Treg) cells to affect late-onset (aged) multiple sclerosis (MS) – with the model of experimental autoimmune encephalomyelitis (EAE)
Authors: Wang Weikan , Thomas Rachel , Oh Jiyoung , Su Dong-Ming 
Published: January 2023

Abstract
A master immunoregulator FoxP3-expressing CD4+ Treg cellsplay an ameliorative role in the severity of MS/EAE disease. However, it is contradictory that the aged T cell immune system possesses relatively enhanced thymic CD4+ Treg generation and accumulated peripheral CD4+ Treg cells, but coupled with severe MS/EAE symptoms and pathology. Using single-cell (sc)-RNA-Seq assay, we found that CNS-infiltrating CD4+ Treg cells in aged EAE mouse model had increased pathogenic properties, showing co-expression of infg and il17a with foxp3, and reduced suppressive effect, exhibiting increased clonal expansion of pathogenic CD4+ T cells. These indicate changed Treg quality in aged EAE mice. Transient inhibition of aged peripheral FoxP3+ Treg cells mitigated the disease severity in the aged mice. The ameliorative effect was more significant when partially inhibiting FoxP3+ Treg cells with a drug P300i than completely depleting FoxP3+ Treg cells in FoxP3DTR transgenic mice. The mitigation is probably attributed to the correction of Treg cell distribution outside and inside the CNS. By inhibiting accumulated FoxP3+ Treg cells adhering to the brain’s choroid plexus (outside the CNS), the IFN-g-producing cells can be restored, thereby, the impediment of immune cell trafficking into the inflamed CNS is released in aged EAE mice. As a result, the proportion of myelin-specific CD4+ Treg cells inside the CNS was increased for repairing neuroinflammatory damage. Together, the underlying mechanism of severe MS symptoms in elderly patients is associated with the accumulation of Treg cells outside the CNS, which prevents the reparative antigen-specific Treg cells from entering the CNS during the disease.
Supported by NIH/NIAID R01AI121147
Estradiol Effects on Polymorphonuclear Cell Production and Actions Contribute to Estrogen-Mediated Lymphangioleiomyomatosis (LAM) Progression
Authors: Minor Briaunna M N , Koudouvoh Jules , Gibbons Erin , Seger Christina , Hammes Stephen 
Published: January 2023

Abstract
The estrogen sensitivity of Lymphangioleiomyomatosis (LAM)—a rare lung disease involving the slow growth of smooth muscle-like adenomas that result in cystic change in the lung parenchyma—is traditionally examined to assess the direct effects of estradiol on the tumor cells. However, we have observed that the estrogen sensitivity of in vitro models is markedly less pronounced than that of in vivo models, suggesting that estradiol may act elsewhere to promote tumor progression. We have previously reported that polymorphonuclear cells (PMNs) are preferentially increased in the immune microenvironment and regulate growth of LAM-like TSC2-null uterine tumors. We hypothesized that, in addition to direct effects of estrogen on tumor cells, estrogen might also stimulate tumor growth by promoting PMN production in the bone marrow and actions in the tumor microenvironment. Here, we show that estrogen availability influences the levels of PMNs in naïve and tumor-burdened mice. Using bone marrow cultures and CFC assay, we demonstrate that estradiol is a potent inducer of PMN production in chronic inflammatory conditions. Employing both pharmacologic agents and estrogen receptor (ER)-null bone marrow, we showed that ER□ is necessary for promoting a PMN fate for myeloid progenitors. While we see that the migratory, invasive, and proliferative capacities of TSC2-null cell lines are not augmented when stimulated with E2 directly, evidence shows estrogen promotes the pro-tumorigenic function of PMNs co-cultured with TSC2-null cell lines. These data provide insight into the robust effect of in vivoestrogen stimulation as estradiol may be a dual effector in LAM tumor progression and great target for anti-LAM therapeutic strategy.
Supported by grants from NIH (R01CA193583, F31 CA254132-01).
Assembly and localization of extracellular matrix protein fibronectin modulates Natural killer cell infiltration in tumor spheroids.
Authors: Rajasekaran Narendiran , House Trenton , Magdaleno Carina Crystal , Varadaraj Archana 
Published: January 2023

Abstract
Natural Killer cells (NK), the major innate effector cells, with their broad cytotoxicity against tumors are ideal candidates for immunotherapy. Once having entered the malignant site, NK cells encounter a complex environment composed of tumor cells, non-tumor cells along with the extra cellular matrix (ECM). As one of the integral components of the tumor microenvironment (TME), the normal regulation of the extra cellular matrix (ECM) is necessary to prevent tumor progression and metastasis. Nevertheless, the impact of the matrix architecture found in solid tumors on immune cells and especially NK cells is not well characterized. Here we investigated the role of Fibronectin, a fibrous ECM protein, and its assembly and localization on NK cell infiltration into clear cell renal cell carcinoma (ccRCC) spheroids. 3D tumor spheroids established from the ccRCC cell line 786-O (RCC-) that lacks the Von Hippel-Lindau (VHL) gene are incapable of fibronectin assembly in their ECM. However, addition of the VHL gene into the 786-O cells (RCC+) reversed this phenotype facilitating fibronectin assembly. Fibronectin in RCC-spheroids exhibited prominent peripheral localization in contrast to RCC+ spheroids where fibronectin was detected towards the center of the spheroid. Coincident with fibronectin localization, PKH26 labeled Natural killer cell line, NKL, showed significantly increased infiltration into RCC+ spheroids compared to RCC-spheroids that lacked the tumor suppressor VHL. Thus, assembly and localization of fibronectin modulates NK cell infiltration in RCC tumors.
Supported by grants from NIH U54MD012388 (NR and AV) and NIH U54CA143925 (NACP-NAU) to NR.
LTi-like type 3 innate lymphoid cells are associated with the transient induction of tertiary lymphoid structures in inflamed pulmonary tissue
Authors: Riffard Clemence , Teillaud Jean-Luc , Dieu-Nosjean Marie-Caroline 
Published: January 2023

Abstract
Tertiary lymphoid structures (TLS) are ectopic lymphoid aggregates observed in inflamed tissues. Their presence in pulmonary tissue has been associated with good prognosis in patients with lung cancer. Although TLS formation shares common features with secondary lymphoid organogenesis, the mechanisms underlying this process are yet to be fully elucidated. Presence of ILC3 in the vicinity of tumor-associated TLS correlates with the density of lymphoid structures. We hypothesized that RORγt+ ILC3 play a major role in TLS induction in inflamed lungs, as do RORγt+ fetal lymphoid tissue inducer cells (LTi) during lymph node formation. We developed a murine model of pulmonary inflammation induced by LPS intranasal instillation that triggers TLS formation. ILC3 presence in lungs was observed with a specific flow cytometry antibody panel. Three RORgt+ ILC3 subsets (NCR− CCR6−, NCR− CCR6+, NCR+ CCR6−) were detected. In particular, NCR− CCR6+ LTi-like cells represented about 25% of CD117+ CD127+ ILC (ILC2/ILC3) and less than 0.04% among CD45+ cells. Analyses of ILC subsets at different time points following LPS instillation showed an increase in the number and % (up to 0.59%) of ILC3 among CD45+ cells, as well as an increase in the % of LTi-like ILC3 among CD45+ cells, that peaked around day 10 (from 0.02% up to 0.15%). Quantification of TLS by histochemistry showed that they are induced in lung tissue in an LPS dose-dependent manner. TLS density increases between days 3 and 10 and decreases thereafter. They were no longer detected at day 25, while LTi-like ILC3 % returned to baseline. Gene expression analysis and functional assays of sorted LTi-like ILC3 are essential to determine if these cells are key players in TLS induction.
Supported by grants from La Ligue Nationale contre le cancer (TAHG21042) and Fondation ARC (R19232DD).
Resident memory T cells in antitumor immunity and cancer immunotherapy
Authors: Mami-Chouaib Fathia , Tihy Isabelle , Corgnac Stephanie 
Published: January 2023

Abstract
Tumor-resident memory T (TRM) cells play an important role in cancer diseases. Accumulating evidence indicates that subpopulations of CD3+ CD8+ tumor-infiltrating lymphocytes (TIL) are TRM cells, and are emerging as activated tumor-specific T cells. These TRM are characterized by expression of CD103 integrin and identify tumor-reactive cytotoxic T lymphocytes. Human lung tumor CD103+ CD8+ TRM co-express CD49a and CD69, and are associated with a favorable prognosis. They also express a panel of T-cell inhibitory receptors, including PD-1, and are able to kill autologous tumor cells upon blockade of PD-1 with neutralizing antibodies. This CD103+ CD8+ TRM subset also emerges as a predictive biomarker of response to PD-1 blockade and expands during anti-PD-1 treatment in responder, but not in non-responder patients. It is now widely admitted that expression of CD103 on activated CD8+ T lymphocytes and persistence of TRM in epithelial tissues requires TGF-β. This cytokine is secreted in the tumor microenvironment in its inactive form bound to latency-associated protein, and is activated by αV integrins on tumor cells and immune cells. Using multiplex immunofluorescence staining on cohorts of anti-PD-(L)1-treated lung cancer patients, we recently showed that decreased expression of tumor αV is associated with improved immunotherapy-related-progression-free survival and correlates with an increased density of CD8+ CD103+ TIL. Cancer cell αV activates autocrine TGF-β and participates in inducing CD103 on activated CD8+ T cells. Our more recent studies in mouse tumor models and human cancers demonstrate that different CD8+ TRM subsets participate to response different immune checkpoint inhibitors and therapeutic cancer vaccines.
Supported by ARC, INCa, Ligue
Supported by ARC, INCa, Ligue
Mucosal vaccination provides protection from HSV-2 infection and disease
Authors: Tucker Kiersten , Dave Veronica , Lund Jennifer M 
Published: January 2023

Abstract
Herpes simplex virus type 2 (HSV-2) is a sexually transmitted pathogen that is estimated to infect around 23 million people per year. Despite high global prevalence, there are not any approved vaccines that are therapeutic or preventative. Most vaccines that we use today rely on injecting antigens intramuscularly in order to elicit an adaptive immune response. However, given that most pathogens gain entry to the host across barrier surfaces, a focus on eliciting mucosal immunity may enhance protection; vaccine-induced local immunity at the site of first pathogen exposure may have the best chance at preventing the spread of infection beyond the pathogen portal of entry. We hypothesized that a mucosal immunization would prime memory T cells to reside in vaginal tissues and provide better protection against vaginal HSV-2 exposure than other routes of immunization. Our initial data suggests that intranasal immunization is effective in protecting mice from vaginal HSV-2 infection. Ongoing work focuses on characterizing the role of vaccine-elicited mucosal CD8+ T cells in preventing infection to provide insight into the mechanisms of protection induced by mucosal immunization for HSV-2. These findings contribute to the efforts to generate an effective vaccine to prevent HSV-2 infection and disease.
Supported by the Mary Gates Research Scholarship and the Art Levinson Award through the University of Washington.