bluuebunny/crossref_metadata_2025_split huggingface dataset:
A critical role for the RNA binding protein HuR in CD4+ T cell-mediated airway inflammation in mouse and human
Authors: Fattahi Fatemeh , Ellis Jason S , Sylvester Michael , Bahleda Kristin , Lugogo Njira L , Atasoy Ulus 
Published: January 2023

Abstract
The RNA-binding protein HuR (elavl1) positively regulates the expression of its targets by increasing mRNA stability and translation. Th2 and Th17 related genes play critical roles in airway inflammation in both mouse and human. Using in vitro and in vivo studies, we have previously shown that these proinflammatory genes are regulated by HuR. Here, we generated a conditional ablation of HuR in T cells (distal Lck-cre HuRfl/fl). We found significant decreases in Th2 differentiation, cytokine production, and lung inflammation in an alum-emulsified ovalbumin (ova-alum) model of allergic airway inflammation using this novel mouse model. We hypothesized that HuR might similarly regulate lung inflammation in human asthma. Using peripheral CD4+ T cells from 45 asthmatic patients, we found the inhibitory effect of the HuR-specific inhibitor CMLD-2, as well as an AMPK activator (acadesine aka AICAR), on both Th2/Th17 signature cytokines in both protein and mRNA levels. Our asthmatics were classified into type 2 high or non-type 2 high based on their blood eosinophil and FeNO levels. Interestingly, there were significant positive correlations between Th2 cytokine levels in CD4+ cells with levels of eosinophils and FeNO from asthmatics. Surprisingly, IFNγ (protein and mRNA) were also markedly decreased by HuR inhibition. Our bioinformatic studies also revealed proteins with high interaction with HuR including MAPK14/p38α and PTEN, common in the pathways with IFNγ, are affected by HuR inhibitor. Taken together, our mouse and human data suggest that HuR plays a crucial permissive role in both allergen- and nonallergen-driven airway inflammation by regulating key genes and that interfering with its function may be a novel way to treat asthma.
Supported by R01: AI080870, R21: AI079341
QRICH1 is a CARD11 interactor that negatively regulates T cell activation
Authors: Carter Nicole M , Pomerantz Joel L 
Published: January 2023

Abstract
Precise regulation of lymphocyte activation is crucial for fighting off pathogens while preventing autoimmunity and lymphoma. CARD11 is a scaffold protein and signaling hub that links antigen receptor engagement to activation of the NF-κB, JNK, and mTOR pathways. We identified QRICH1 as a previously unrecognized binding partner of CARD11 that binds CARD11 in a signal-dependent manner during T cell receptor (TCR) signaling. To identify the function of QRICH1 in TCR signaling, we used CRISPR/Cas9 genomic editing to disrupt QRICH1 expression in Jurkat T cells. QRICH1 knockout resulted in enhanced TCR-induced NF-κB activation and IL-2 secretion, while QRICH1 overexpression in HEK293T and Jurkat T cells dose-dependently inhibited NF-κB activation by wild-type CARD11 and by constitutively active and oncogenic CARD11 mutants. Domain deletion experiments revealed that the inhibitory activity of QRICH1 resides in the N-terminal half of the protein, while the C-terminus contains an autoinhibitory domain that participates in intramolecular interactions and modulates the ability of QRICH1 to inhibit CARD11 signaling to NF-κB. To study the function of QRICH1 in vivo, we generated mice with a conditional QRICH1 allele. We found that QRICH1-deficient CD4+ and CD8+ T cells produced higher levels of IL-2 and IFN-ɣ, respectively, following stimulation by anti-CD3/CD28. Our findings identify QRICH1 as a negative regulator of T cell activation that modulates the signaling output of CARD11.
This work was supported by NIH grants F31CA254167, RO1AI148143, and T32GM007445.
Malnutrition Disrupts T Cell Migration
Authors: Dadzie Kwesi A , Foster Takesha R , Mister Abigail , Goulmamine Syreen , Gibson David , Adams Olivia , Gubbels-Bupp Melanie R 
Published: January 2023

Abstract
Malnutrition is associated with reductions in the number and function of T lymphocytes. Previous studies in the lab suggest that malnutrition may also impart a “super-quiescent” phenotype to T cells, perhaps affecting the efficiency of their migration within and between lymph nodes. Thus, the purpose of this study is to evaluate the effect of malnutrition on T cell migration in vivo and to characterize malnutrition-induced changes in the expression of proteins known to be important for T cell migration. To determine if malnourishment alters T cell migration in vivo, we compared lymph node entry rates of adoptively-transferred malnourished and control T cells in malnourished and control recipients. In agreement with other studies, control CD4+ T cells were more efficient than control CD8+ T cells at entering the lymph nodes. Interestingly, regardless of recipient diet, malnourished CD4+ and CD8+ T cells entered the lymph nodes at equivalent rates, suggesting that malnourishment eliminates distinct lymph node entry efficiencies for CD8+ and CD4+ T cells. We also found important differences in the expression of key proteins involved in T cell migration between malnourished and control mice. Overall, we found that malnutrition disrupts T cell migration including the distinct migration efficiencies of CD4+ and CD8+ T cells. An improved understanding of T cell-intrinsic changes that occur during malnourishment should enhance our knowledge of CD4+ and CD8+ T cell migration and shed light on how organisms adapt to malnutrition.
Supported by NSF-MRI [DBI- 1920116] NSF-RUI [IOS-1951881]
IL-4 Signaling Regulates the Fate of B Cell Differentiation and limits BCR repertoire
Authors: Oviedo Juan Marcos , Gibbs Lisa C , James Kimra , Fairfax Keke C 
Published: January 2023

Abstract
Il-4 has a critical role in organizing the peripheral lymph nodes in homeostasis and after vaccination. To further understand the molecular mechanisms and their impact in the immune response, we used single cell RNA-Seq of CD45+ cells in 4get and 4getIL4R−/− mice following tetanus/diphtheria immunization. We found that the lack of IL-4 signaling restrains the expression of key co-stimulatory and differentiation molecules CD83, CD86, and xbp1, while upregulating the jun/junb/fos pathways. These alterations result in reduced B cell proliferation and differentiation into plasma cells. Surprisingly, we found that in the absence of IL-4 signaling the ratio of dark to light zone residing germinal center shifts from the normal ratio to one dominated by the dark zone. The increase in Dark Zone germinal centers lead to an increase in the diversity of the repertoire in vaccinated 4getIL4R−/− mice as confirmed by VDJ-seq. We also observed that the upregulation of the transcription factor T-bet has the capacity of inducing a similar phenotype. Finally, even though the amount of memory B cells was increased in the absence of IL-4 signaling, we observed that these cells are not able to respond to a secondary challenge. These data suggest a pivotal role for IL-4 in governing B-cell differentiation and the development of optimal cellular and humoral immunity to vaccination.
supported by grants from NIH R01 AI135045
Lineage specification of killer versus helper Innate lymphoid cells
Authors: Ding Yi , Lavaert Marieke , Harly Christelle , Das Arundhoti , Bhandoola Avinash 
Published: January 2023

Abstract
Helper innate lymphoid cells (ILCs) combat pathogen infection by rapid secretion of cytokines whereas killer ILCs (NK cells) eliminate virus-infected cells or tumor cells during early defense. However, the early development of NK cells and lineage relationships between NK cells and helper ILCs remain elusive. In PLZF lineage tracing model, normal NK cells are traced at a much lower ratio than helper ILCs, which indicates most NK cells develop from progenitors that do not express PLZF. However, it is also possible that most NK cells develop from PLZF-expressing ILC progenitors (ILCPs), but PLZF-labelled NK cells may proliferate less than unlabeled NK cells. To address these possibilities, we transferred ILCPs and its upstream progenitors early innate lymphoid progenitors (EILPs) and all lymphoid progenitors (ALPs), and then assessed PLZF labelling in NK cells derived from these progenitors. We found that ILCP-derived NK cells were traced at a ratio as high as in helper ILCs whereas EILP and ALP-derived NK cells were labelled at a much lower ratio, which is similar to the ratio in normal NK cells. These results indicate that NK cells develop from both ILCP-dependent and ILCP-independent pathways. Our findings establish novel lineage relationships between NK cells and helper ILCs. We are currently assessing whether different developmental pathways of NK cells lead to functionally distinct subsets of NK cells.
Sympathetic nervous system-mediated β-adrenergic signaling maintains the pool of mature natural killer cells
Authors: Geyer Rory , Moussa Marmar , Mandoiu Ion , Srivastava Pramod K , Nevin James 
Published: January 2023

Abstract
The sympathetic nervous system (SNS) innervates all lymphoid tissues and is, thus, well-positioned to influence the activity of immune cells. Accordingly, the coordinated responses of the SNS and immune system are crucial for immune responses to infection and cancer. Natural killer (NK) cells are essential for the immune response to certain viral infections and cancers in which they kill infected or cancerous cells and produce cytokines and chemokines to instruct the activity of other cells contributing to the response. We find here that steady state SNS signaling maintains the balance of mature and immature NK cells in the spleen and bone marrow. Using 6-hydroxydopamine to ablate sympathetic nerve terminals, we show by single cell RNA-sequencing and flow cytometry that SNS-ablated mice have decreased frequencies of mature CD27− CD11b+ NK cells and increased frequencies of immature CD27+ CD11b− and intermediate CD27+ CD11b+ NK cells. An increased ratio of immature NK cells to mature NK cells has been linked to decreased anti-tumor immunity. Along these lines, SNS-ablation also increases the growth rate of RMA-S tumors, the rejection of which depends on NK cells. This accelerated tumor growth was dependent on the absence of both α- and β-adrenergic signaling. Culture of early NK cell precursors in the presence of selective α- or β-adrenergic agonists suggests that signaling through β-adrenergic receptors promotes the maturation of NK cells. These data suggest that signaling through β-adrenergic receptors mediated by norepinephrine produced by the SNS maintains the pool of mature NK cells, allowing them to effectively survey peripheral tissues and participate in anti-tumor immunity.
Exposure to diverse microbial flora accelerates the development of infant immunity
Authors: Schuldt Nathan J , Burger Sarah , Stenger Terran , Pierson Mark , Huggins Matt 
Published: January 2023

Abstract
The newborn immune system is considered underdeveloped and ill-equipped to defend itself against many pathogens. Indeed, infection is the leading cause of hospitalization and death for infants under 6 months. Relative to the adult immune system, little is known about infant immunity or the signals that orchestrate immunologic maturation. The constraints of human research and the use of sterile animal models limit advancements in this area. Commonly used animal models are raised in “specific pathogen free” (SPF) conditions to minimize the confounding effects of diverse microbial exposure. We sought to understand the physiologic effects of exposure to those diverse microbes on infant immune development. We co-housed laboratory mice with pet store mice from conception to expose them to diverse microbial flora. Microbial exposure led to a dramatic shift in immune cell composition with increases in the proportion of T cell memory, helper T cell subsets, macrophages, and neutrophils, as well as general increases in the number of most immune cell subsets as early as 2 weeks of age. The gap between the microbially-exposed mice and SPF mice continued to widen at 3 and 6 weeks of age. We also observed increased TNFα, IL-6, IL-5, IL-17, and RANTES expression at 3 weeks of age. By 6 weeks most of the cytokines/chemokines tested where increased. These observations demonstrate that exposure to diverse microbial flora during early life dramatically alters the trajectory of the developing immune system. A better understanding of how healthy immune systems develop, and the risks and benefits of modulating infant immunity could lead to important advancements in treatments for early life infection and novel infant vaccination approaches.
Supported by Grants from: Medical School/University of Minnesota Foundation Award Program Masonic Cross-Departmental Grants in Children’s Health Research University of Minnesota Department of Pediatrics “R” Award
Neonatal Immunity to Malaria using a mouse model
Authors: Opata Michael M , Gbedande Komi , Smith Margaret Rose , Onjiko Robert Sifa 
Published: January 2023

Abstract
High mortality rates caused by malaria are seen in children under five years, but there is limited knowledge on neonatal immunity to the Plasmodium infection due to the lack of a reliable neonatal animal model to study the disease in children. By utilizing a newly developed young rodent model in our lab, we have observed that purified CD4+ T cells from day 14 old young mice (pups) proliferate slower than those purified from 8-week adult mice when stimulated in vitro. To investigate neonatal CD4+ T cell response to malaria, we infected 14-day old pups or adult mice with Plasmodium chabaudi and determined the activation status (CD127− CD44+ CD11a+) and differentiation of effector (Teff) subsets, using the expression of CD62L or CD27. Pup CD4+ T cells were activated by downregulating CD127, even though in lower proportions and numbers when compared to adult CD4+ T cells. Surprisingly, there were more proportions of TeffEarly (CD27+ CD62L+) and TeffInt (CD27+ CD62L−) subsets in the pups than adult mice, which had mostly TeffLate (CD27− CD62L−), suggesting that the transition of Teff subsets is slower in the young mice. We next determined cytokine secretion and Tbet expression by the Teff. Apart from IL-2 which was high in the pups, there was no difference in the proportions of protective cytokines including IFNγ, IL-10 and TNFα in both groups. Effector CD4+ T cells from the pups expressed high proportions of the Tbet transcription factor when compared to adult cells. Taken together, these data suggest that pup CD4 T cells may require more Tbet to differentiate into functional effector cells during Plasmodium infection.
Supported by Depart of Biology Appalachian State University
Th1/Tfh hybrid (Ifng+Il21+) T cells contribute to delay of germinal center formation in Plasmodium chabaudi infection
Authors: Clark Lucinda Puebla , Gbedande Komi , Domingo Nadia , Stephens Robin 
Published: January 2023

Abstract
Germinal center (GC) formation in P. chabaudi rodent malaria is delayed for three weeks, which can prolong the infection. T follicular helper (Tfh) cells express CXCR5 for homing to the B cell area, and IL-21 which promotes antibody production. T helper type 1 (Th1) cells express CXCR3 and IFN-γ to promote phagocytosis. Previous data suggests that Th1 responses control peak parasitemia while systemic Th1 cytokines inhibit GC formation. In P. chabaudi, majority of CD4+ effector T cells (Teff) express markers of both Th1 and Tfh (e.g., CXCR5, CXCR3, Th1/Tfh hybrid), including IFN-γ and IL-21. The contribution of Th1/Tfh hybrid T cells in protection and antibody production remains unclear. We tested the impact of Il21+ Tfh-like, Ifng+ Th1-like, and Ifng+ Il21+ Th1/Tfh in protection by transferring these subsets from infected triple cytokine reporters (Il21-rfp Il4-gfp Ifng-Thy1.1) into TCR KO mice that are then infected. We are currently assessing changes in parasitemia and pathology. We found that Th1/Tfh cells do not help in vivo like recent data suggest they do in vitro. Th1/Tfh significantly generated few GC B cells, compared to Il21+ Tfh-like cells. Recipients of Th1-like cells had significantly lower parasite-specific IgG and GCs by immunohistology than Tfh-like at 21 dpi, hybrid Th1/Tfh were intermediate. A cytokine kinetics study of Th subsets in infected cytokine reporter mouse distinguished Il4+ Il21+ GC Tfh from Th1 cells. The inflammation-driven chemokine receptors CXCR3 and CXCR6 are highly expressed on GC Tfh and Th1/Tfh hybrid during the first week of infection, but decline as GCs develop, suggesting a role for T cell migration which could explain the delay in GC formation and parasite-specific antibody production in malaria.
Supported by R01AI 135061
A forward genetic screen in alveolar macrophage-like cells highlights the essential role of peroxisome membrane regulation in hallmark alveolar macrophage phenotypes
Authors: Thomas Sean M , Wierenga Kathryn , Pestka James , Olive Andrew J 
Published: January 2023

Abstract
Alveolar macrophages (AMs) are a tissue resident macrophage population within the lung that are critical to maintain lung homeostasis and respond to inhaled antigens. Mechanistic investigations of AM function employing genetic approaches like CRISPR-Cas9 are hindered by the inability to obtain large numbers or maintain the AM-likeness of these cells ex vivo. We recently developed fetal liver-derived alveolar-like macrophages (FLAMs), a model that maintains the expression profile and immune functions of AMs long-term ex vivo. Importantly, FLAMs are genetically tractable using CRISPR-Cas9, enabling previously impossible interrogation of AM functions. Leveraging this innovative tool, we generated a genome-wide knockout FLAM library and are now completing forward genetic screens to dissect the regulation of AM maintenance and function. One such screen identified genes that regulate the surface expression of the AM-specific protein, Siglec-F. Among the numerous AM-associated pathways we identified as positive regulators of Siglec-F, the most robust signature was for genes related to the peroxisome. Thus, we hypothesize that the peroxisome is an essential signaling platform for AM functions. To test this prediction further we are using chemical and genetic approaches to dissect peroxisome functions in AMs. We are examining how peroxisomes modulate gene expression and metabolic flux in AMs and determining how this alters AM responses to stimuli. Together, these results highlight the utility of our novel genetic screening platform in FLAMs to unlock our understanding of AM functional mechanisms like never before.
This work was supported by the Rackham Research Endowment Award from Michigan State University.
Cbf-β is required for development and differentiation of murine mucosal-associated invariant T cells
Authors: Toor Jugmohit , Liu Tingting , Subedi Kalpana , Wang Jie , Loveless Ian , Yin Congcong , Zhou Li , Mi Qing-Sheng 
Published: January 2023

Abstract
Mucosal-associated invariant T (MAIT) cells primarily reside in mucosal tissues and tissues exposed to the environment to act as a defense against microbial threats. Additionally, they are also involved in a broad spectrum of diseases, including infection, cancer, allergy, and autoimmunity. Although some transcriptional factors and micro-RNA have been recently identified to regulate MAIT cell development and differentiation, a significant gap in our knowledge of MAIT cell regulation still exists. The core binding factor subunit beta (Cbf-β) is a non-DNA binding protein forming a heterodimer with RUNX family proteins to regulate diverse signaling pathways which maintain homeostasis of a wide range of immune cells. To understand the role of Cbf-β in MAIT cell development and function, we generated T cell lineage specific Cbf-β knockout mice (CD4cre+Cbf-β KO) and inducible Cbf-β knockdown mice (UBCcre+Cbf-β KO). We found that Cbf-β deficiency impaired thymic MAIT cell development and interrupted MAIT1 and MAIT17 differentiation. Additionally, inducible knockdown of Cbf-β reduced activation and cytotoxicity in peripheral MAIT cells. Our work suggests that Cbf-β is required for thymic MAIT cell development and MAIT1 and MAIT17 commitment in addition to playing a role in MAIT cell activation and cytotoxicity. Overall, Cbf-β may serve as a novel target to modulate MAIT cells in MAIT cell-based immunotherapies.
This study was supported by Henry Ford Immunology Program grants (T71016 and T71017) to QSM and LZ, by the National Institutes of Health (NIH) grants R61AR076803, R01AR063611, and R01AR069681 to QSM, RO1AR072046 to LZ.
Modulating expression of a BTB-ZF transcription factor in adipose resident NKT cells alters function and disrupts metabolism regulation.
Authors: Millick Danielle , Vieth Joshua A , Denzin Lisa K , Sant’Angelo Derek B 
Published: January 2023

Abstract
Natural killer T cells (NKTs) are potent modulators of the immune system that can regulate the function of other cell types largely through their rapid and robust production of a wide variety of cytokines. One tissue that relies on the interactions of NKT cells with various cell types is white adipose tissue. Adipose resident NKT cells (arNKTs) maintain tissue homeostasis and metabolism through anti-inflammatory interactions such as modulating macrophage functions and enhancing the proliferation and function of FoxP3-expressing Tregs. However, arNKTs are phenotypically distinct from other populations of NKT cells in their transcription factor expression. We have found that genetically modulating the expression of a BTB-ZF transcription factor in NKT cells alters the frequency and function of arNKTs, thereby altering weight gain and metabolic function. Our data show that sustained expression of the BTB-ZF transcription factor in arNKT cells leads to lower arNKT frequency, altered function, and enhanced weight gain in a model of obesity (60% high fat diet). While deletion of the transcription factor during NKT cell development leads to lower arNKT frequency, altered arNKT phenotype, and impaired weight gain when fed 60% high fat diet. Collectively, our data show that dynamic changes in the expression of a BTB-zinc finger transcription factor during NKT cell development and tissue-homing are required for normal arNKT cell function, and preservation of adipose tissue metabolism. Our findings imply that NKT cell inability to temporally regulate the expression of a single transcription factor may drive adipose tissue dysregulation and metabolic disease.
Supported by NIH/NIAID R21 AI159066-01
Cell-interaction based CRISPR screening using intercellular barcode transfer
Authors: Tay Neil Q , McManus Michael 
Published: January 2023

Abstract
Cell to cell interaction is one of the fundamental paradigms in cell biology and it is central to immune processes. While recent advances have allowed us to monitor cell interactions both in vitro and in vivo, doing so at large scales with complex interactions remain challenging. We have developed Relay, a scalable technology by which cellular interactions can be tracked via molecular barcoding and intercellular transfer of barcodes. We employed our system to perform a novel cell interaction-based CRISPR screen that was not possible with conventional CRISPR screening strategies. In our screen, we investigated factors that are involved in antigen processing, presentation, and transfer in myeloid cells using a myeloid-T cell co-culture system. By utilizing CRISPR/Cas9 together with Relay, we were able to perform genome-wide perturbations in myeloid cells and concurrently track myeloid-T cell interactions, allowing us to directly associate antigen sampling by T cells to genetic perturbations affecting antigen processing in the myeloid cells.
Supported by grant from NIH (U01 CA217882).