bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bhlhe40 maintains CD8+ T cell fitness and functionality in non-lymphoid tissues and tumors
Authors: Li Chaofan , Taneja Reshma , Sun Jie 
Published: January 2023

Abstract
Recent evidence has suggested that tissue-resident and tumor-infiltrating CD8 T cells share common molecular signatures for their maintenance in the non-lymphoid environment. Currently, the molecular mechanisms regulating the residency, health and function of effector and memory CD8 T cells in non-lymphoid tissue remain largely elusive. Here we have identified that the transcription factor Bhlhe40 is critical for the fitness and function of effector & memory CD8 T cells in the non-lymphoid tissue and tumor. Bhlhe40 and its associated gene signature are highly expressed in CD8 T cells, which reside in non-lymphoid tissues and tumor, compared with their counterparts in the spleen following viral infection and tumor transplantation. We find that Bhlhe40 deficiency results in the progressive loss of effector and memory CD8 T cells in non-lymphoid tissues following influenza infection and tumor transplantation. As such, Bhlhe40 abolition leads to defective anti-cancer immunity and impaired heterologous anti-viral immunity mediated by the tissue-resident effector and memory CD8 T cells. Mechanistically, we show that Bhlhe40 is dispensable for the activation and metabolic programming of CD8 T cells, but is vital for them maintaining metabolic and mitochondrial fitness. As such, Bhelhe40 deficiency results in energy failure of CD8 T cells and impaired maintenance of active histone marks in the loci of CD8 T cell effector molecules. Therefore, we have identified Bhlhe40 as a key TF for the maintenance of CD8 T cell fitness and functionality in non-lymphoid tissues. Furthermore, our data suggest that the manipulation of Bhlhe40 expression and function may serve as a potential strategy to boost the efficacy of vaccination and immunotherapy.
TRAF3-mediated regulation of the T cell receptor complex
Authors: Arkee Tina , Wallis Alicia M. , Bishop Gail A. 
Published: January 2023

Abstract
TRAF3 is an adaptor protein with diverse cell type-specific functions that plays an important role in T cell biology. T cell conditional TRAF3−/− mice, despite normal numbers of T cells, have poor responses to immunization and infection. Further investigation revealed that early events in T cell receptor (TCR) signaling are defective in TRAF3-deficient T cells. Examining the basis of this defect, we recently reported that TRAF3 normally inhibits localization of the negative regulators Csk and PTPN22 to the TCR membrane complex. We now wish to understand how TRAF3 is recruited to this complex. TRAF3 associates with the TCR-CD28 complex upon stimulation; engagement of both the TCR and CD28 is required for TRAF3 recruitment. We hypothesize that TRAF3 associates with the TCR complex either indirectly, by binding to other CD28-associated adaptor proteins, or directly, by binding to motifs in the cytoplasmic tail of CD28. Results of preliminary experiments suggest that the TRAF3 TRAF-C domain associates with Linker of Activated T cells (LAT), but not with the LAT-binding adaptor protein Gads. We are currently investigating the structural requirements for the association of TRAF3 with CD28, the adaptor protein Grb2, and other TCR signaling proteins, to determine how TRAF3 is recruited to the TCR complex. Based upon our previous finding that TRAF3 enhances TCR signaling by inhibiting the localization of negative regulatory proteins associated with the TCR, as well as our preliminary data indicating that TRAF3 interacts with members of the LAT complex, we are also investigating the interactions between TRAF3 and negative regulatory proteins associated with the LAT complex.
The transcription factor T-bet is regulated by microRNA-155 in murine anti-viral CD8+ T cells via SHIP-1.
Authors: Hope Jennifer L. , Stairiker Christopher J. , Spantidea Panagiota I. , Gracias Donald T. , Carey Alison J. , Fike Adam J. , van Meurs Marjan , Brouwers-Haspels Inge , Rijsbergen Laurine C. , Fraietta Joseph A. , Mueller Yvonne , Klop Rosemarieke C. , Stelekati Erietta , Wherry E. John , Erkeland Stefan , Katsikis Peter D. 
Published: January 2023

Abstract
MicroRNAs (miRNAs) are small, single-stranded non-coding RNAs that play essential roles in regulating key cellular processes, and are highly conserved across species. However, the regulatory role of microRNAs in generating effective CD8+ T cell (CTL) responses to viral infection and tumors is only now being elucidated. Microarray miRNA expression profiling identified unique expression patterns of miRNAs in naïve and effector anti-viral CTL. In particular, microRNA-155 was significantly upregulated in effector CTL. Previously, we have demonstrated that miR-155 is essential to CD8+ T cell responses to viral and intracellular bacterial infections. In the present study, we show that miR-155 overexpression in CTL augments anti-viral effector CTL and skews memory CD8+ T cells towards an effector memory phenotype. MiR-155 overexpression resulted in enhanced T-bet expression, which is known to be required for effector CTL and to promote effector memory cell formation. Importantly, we show that the proliferative effect of miR-155 on CD8+ T cells is mediated by T-bet. In CTL, T-bet levels were controlled in vivo by miR-155 via SH2 (Src homology 2)-containing inositol phosphatase-1 (SHIP-1), a known direct target of miR-155. Our studies have revealed an important and unexpected link between miR-155, T-bet transcription factor and SHIP-1 in the context of CTL responses to an in vivo viral infection.
The IRF4 gene regulatory module functions as a read-write integrator to dynamically control T helper cell fate
Authors: Sciammas Roger , Krishnamoorthy Veena , Kannanganat Sunil , Maienschein-Cline Mark , Cook Sarah , Chen Jianjun , Bahroos Neil , Corse Emily , Chong Anita S. F. 
Published: January 2023

Abstract
Transcriptional regulation during CD4+ T cell fate decisions enables their differentiation into distinct states, guiding immune responses towards antibody production via Tfh cells or inflammation by Teff cells. Tfh–Teff fate commitment is regulated by mutual antagonism between the transcription factors Bcl6 and Blimp-1. Here we examined how T cell receptor (TCR) signals establish and arbitrate Bcl6–Blimp-1 counter-antagonism. We found that the TCR-signal induced transcription factor IRF4 is essential for the differentiation of Bcl6-expressing Tfh and Blimp-1-expressing Teff cells. Increased TCR signaling raised IRF4 amounts and promoted Teff fates at the expense of Tfh ones. Importantly, orthogonal induction of IRF4 expression redirected Tfh fate trajectories towards those of Teff and this occurred independently of IL-2 signals. Mechanistically, we linked greater IRF4 abundance with its recruitment towards low affinity binding sites within Teff cis-regulatory elements, including those of Prdm1. We propose that the Irf4 locus functions as the “reader” of TCR signal strength, in turn, concentration-dependent activity of IRF4 “writes” T helper fate choice.
Hands on, Minds on: How to create effective, student-driven learning without becoming overwhelmed
Authors: Lukin Kara R. , Aviszus Katja 
Published: January 2023

Abstract
When done effectively, student-driven approaches to learning increase student satisfaction and learning outcomes. In addition, encouraging students to be more actively involved in their learning better prepares them for graduate school and for joining the professional workforce. For those used to a more traditional lecturing role in the classroom, the prospect of developing and integrating approaches that require a higher degree of engagement of students can be intimidating. Here, we share our experiences and provide examples of simple strategies and approaches we adopted to implement a classroom environment that requires students to be more actively involved in the learning process. We demonstrate how to create and implement the following methods slowly into courses with any size enrollment:
1) content organizers and hierarchy maps that scaffold concepts and drive students to integrate material, 2) distributive recall practices that increase neural networking, 3) online resources for content mastery so class time can be for application and group activities, 4) IF-AT (immediate feedback) scratch cards and problem sets that drive discussion and learning better than clickers, 5) activities that explain immunologic concepts with which students often struggle, and 6) mini and full case studies that are woven through the term.
All resources will be available for immediate use.
The prevalence of digital note-taking trends in professional pharmacy students
Authors: Hussein Mohamed , Lana Angelica , Reda Betlhem , Bizuayehu-Mekonnen Menorwork , Nassan Motaz , Fazal Nadeem 
Published: January 2023

Abstract
The use of technology in the classroom may have positive and negative effects on student learning. We determined the use of note taking technologies to precept students learning. The research in question is to find out the use of integration of modern technology into didactic courses of pharmacy education. We surveyed students’ perceptions of note taking regarding the type, frequency, and appropriateness of use of note-taking technology in the classroom. We carefully prepared 10 questions for an anonymous survey pertaining to students’ perception of habits and ways of taking notes in the class and how it affected their learning capability in the school. A questionnaire survey was delivered to 59 P1 students in the classroom. 100% of the students who responded owned electronic device and use it while in school. The data determined the following trends. When students were asked about which operating system they are using, 39% used MacBook, 34% used windows and 21% reported use of Ipad for their notes-taking needs. 60% of the students either used a digital product to take notes or used windows. 71% of students who responded said that they take 75-100% of their notes on electronic devices in the P1 courses. The majority of those students used a laptop to take notes during class, 60% of students reported using Notability for note taking while in classrooms. Among other digital products PDF writer, Evernote, and OneNote were also used. 23% of the students still prefer taking notes the old traditional way by printing the lectures and writing notes on paper. This study demonstrated how modern digital technology helps and enhances note-taking ability among pharmacy students. Price and easy use of technological gadgets influences the final selection for students.
The Role of Food Allergy Education in Schools: Measuring Attitudes, Beliefs, and Knowledge
Authors: Canon Nicole , Gharfeh Maya , Anvari Sara , Davis Carla 
Published: January 2023

Abstract

Introduction
Almost 6 million children suffer from food allergies with roughly 2 affected per classroom. However, deficiencies in knowledge and discrepancy in attitudes within school staff when addressing food allergies are associated with barriers to care. In this study, school personnels’ knowledge, beliefs, and attitudes were measured before and after a food allergy educational workshop.

Methods
Six schools in Houston Independent School District were randomly assigned into an interventional (n=4) and control group (n=2). Seven-hundred and thirty-three personnel matched for age, socioeconomic status, ethnicity, and educational level completed the Chicago Food Allergy Research survey before and after a one-hour didactic session. Responses were measured using frequencies and percentages. The group score differences and survey question comparisons were measured with a linear mixed effects model.

Results
Post-test, the intervention group had knowledge scores 19.58% higher than control (95% CI=16.62, 22.53; p&lt;0.001) with no differences pre-test. Odds of agreeing that injectable epinephrine is important was 874 times higher in the interventional schools post lecture. Within the interventional group, personnel were 207 times more likely to agree to an epi-pen for children post-education.

Conclusion
A didactic session improved knowledge and attitudes in personnel in the intervention schools. Given the growing prevalence of food allergy, the emphasis on food allergy education is crucial to allow for familiarization of the condition, early recognition of anaphylaxis, and promotion of injectable epinephrine use.
IL-18 from Batf3-dependent cells licenses natural killer cell IL-10 production during Listeria monocytogenes infection
Authors: Clark Sarah E. , Schmidt Rebecca L. , McDermott Daniel S. , Lenz Laurel L. 
Published: January 2023

Abstract
Natural killer (NK) cells are innate lymphoid cells that regulate the immune response to infection through secretion of both pro- and anti-inflammatory cytokines. The pathogen Listeria monocytogenes (Lm) capitalizes on NK cell production of the anti-inflammatory cytokine interleukin (IL)-10 during establishment of severe infection. Lm-stimulated NK cell IL-10 limits immune cell recruitment and activation, allowing for bacterial expansion. Here, we report that IL-18 from dendritic cells (DCs) promotes this IL-10 production. IL-18 acts directly on NK cells to license IL-10 secretion that is independent of IL-12 and STAT4, which co-stimulate IFNγ secretion during Lm infection and can induce NK cell IL-10 in other contexts. DC release of IL-18 is driven by the Lm p60 virulence protein and requires DC expression of nlrp3 and batf3. Mice lacking nlrp3, il18, il18R, or batf3 fail to accumulate serum IL-10 and are highly resistant to systemic Lm infection. Our data thus show that during systemic infection Lm selectively targets Baft3-dependent cells to drive IL-18 release that licenses NK cell IL-10 production. Exploiting this previously-unappreciated pathway to promote IL-10 production enables Lm to dampen inflammatory and anti-microbial host responses.
Innate-like activation of mucosal-associated invariant T cells in mycobacterial infection
Authors: Huang Shouxiong , Sharma Manju , Zhang Shuangmin , Niu Liang , Zhang Xiang 
Published: January 2023

Abstract
Unlike conventional T cells, mucosal-associated invariant T (MAIT) T cells respond to microbial riboflavin precursor metabolites presented by major histocompatibility complex (MHC)-like molecule MHC-related protein 1 (MR1). Recently, a high percentage of CD8+ T cells reactive to Mycobacterium tuberculosis in humans were identified as MAIT cells instead of conventional MHC class I-restricted T cells. To understand the activation mechanism and effector functions of MAIT cells in mycobacterial infections, we used mycobacterial-incubated human dendritic cells and MR1-overexpressed cells to stimulate primary and clonal human MAIT cells. As a result, the enzyme-linked immunospot assay showed a quick activation kinetics of MAIT cells within hours of mycobacterial stimulation in an MR1-dependent manner. Flow cytometry analysis demonstrated that mycobacterial-incubated antigen presenting cells quickly stimulated primary human MAIT cells to develop a dominant CD69+CD26+phenotype. Activated MAIT cells displayed an enhanced expression of cytokines TNFa, IFNg, IL-17, and granulysin, demonstrating a potential to induce pro-inflammatory and cytolytic responses. Further transcriptomic analysis of the CD69+CD26+subset revealed an integrated activation pathway from conventional CD8+ T cells and natural killer (NK) cells. Especially, activated MAIT cells showed an enhanced gene expression of mitogen-activated protein kinase (MAPK) signaling molecules, NF-kB, T-bet, RORg, and PLZF transcription factors, and pro-inflammatory cytokines and cytolytic molecules. Taken together, our data support that MAIT cells are quickly activated through an innate-like activation mechanism to induce anti-mycobacterial responses.
CHARACTERIZATION OF INTERLEUKIN-17 PRODUCING CELLS IN THE PERIPHERAL BLOOD OF PATIENTS WITH TUBERCULOSIS
Authors: Ruiz-Sánchez Bibiana Patricia , Zarate David Cruz , García-Paredes Victor Gabriel , Hernández-Solis Alejandro , Sabido Raúl Cisero , Serafín-López Jeanete , Estrada-Garcia Iris , Baeza Isabel 
Published: January 2023

Abstract
Tuberculosis (TB), an infection caused by Mycobacterium tuberculosis, is one of the ten main causes of death by infectious disease. When the mycobacteria are inhaled, they interact with immune cells in the respiratory tract in a specialized structure known as granuloma. Interleukin 17 (IL-17) contributes to formation and size of the granuloma, and it correlates with the recruitment of neutrophils. It has been demonstrated that other innate lymphocytes, such as Tγδ cells, contribute to the production of IL-17 during TB. Innate lymphoid cells (ILCs) are other innate lymphocytes that are classified into groups (ILC1, ILC2 and ILC3) according to their cytokine patterns. ILC3 produce IL-17, but it is not known if these cells participate in the response to TB infection. We evaluated, by flow cytometry, the frequency and IL-17 production of Th, Tgd, NK and ILCs (ILC1, ILC2 and ILC3) in the peripheral blood of healthy individuals (negative and positive for the Quantiferon [QTF] test, which detects latent TB), and of patients with active pulmonary TB. The numbers of Th cells, NK cells and ILCs (ILC1, ILC2 and ILC3) were decreased in TB patients, compared to healthy individuals. Peripheral blood was stimulated with M. tuberculosis soluble extract (MTSE) for 5 h in the presence of brefeldin A. After stimulation, Th cells from QTF-positive healthy individuals and TB patients produced IL-17. Tγδ cells also produced IL-17, although the percentage of these cells did not change after activation with MTSE. In some cases, NK, ILC2 and ILC3 from TB patients produced IL-17, but this production did not change after MTSE activation. These results indicate that Th and Tγδ cells contribute to IL-17 production during TB.
This project was funded by SIP-IPN-México (20180254).
Neutrophil extracellular traps deficiency aggravates Citrobacter rodentium infection and the associated intestinal inflammation
Authors: Saha Piu , Yeoh Beng San , Wang Yanming , Vijay-Kumar Matam 
Published: January 2023

Abstract
Peptidyl arginine deiminase-4 (PAD4) is required for the formation of neutrophil extracellular traps (NETs), which is part of the host antimicrobial innate immune response. However, the role of NETs in the pathogenesis and clearance of Citrobacter rodentium (a well-characterized mouse enteropathogen) infection remains unclear. In this study, we challenged the PAD4-deficient (Pad4KO; NETs-deficient) and WT littermates with C. rodentium. Luminal colonization of C. rodentium in Pad4KO mice peaked between 11-14 days post-infection (pi), whereas WT mice cleared the infection with no detectable level of the pathogen in fecal shedding after 7-10 days. Bacterial dissemination to spleen and mesenteric lymph node was significantly higher in Pad4KO mice at day 8 and 28 pi compared to WT. Additionally; Pad4KO mice displayed splenomegaly, colomegaly and elevated colonic and systemic pro-inflammatory markers (lipocalin 2 and serum amyloid A). Histological analysis demonstrated that transmissible colonic hyperplasia and goblet cell depletion were more pronounced in Pad4KO mice. Pad4KO mice also exhibited epithelial barrier dysfunction as indicated by their heightened serum immunoreactivity to bacterial proteins. Flow cytometric analysis revealed a substantial reduction in IL-10-producing macrophages and Th17 cells in the lamina propria and spleens of Pad4KO mice compared to WT mice. However, colonic expression of IL-17A and IL-22 were highly elevated in Pad4KO mice on day 8 pi, but were attenuated on day 28 both in Pad4KO and WT mice. Taken together, our findings highlight that NETs are indispensable in promoting clearance of C. rodentium and resolving the associated intestinal inflammation.
Key role for CD103+ DCs in activating lung iNKT cells during Streptococcus pneumoniae infection
Authors: Crosby Catherine Marie , Mikulski Zbigniew , Kronenberg Mitchell 
Published: January 2023

Abstract
Invariant natural killer T cells (iNKT) are profoundly important for protection against Streptococcus pneumoniae. Prior work has shown iNKT cells are rapidly activated in a CD1d-dependent manner, however the critical antigen presenting cells remain unknown. We have begun to uncover a dominant role for a more recently described iNKT cell subset: NKT17 cells. Within 14 hours of infection, approximately 60% of NKT17 cells have received a TCR signal, and 45% produce IL-17A. Notably, only a minor percentage of NKT1 cells are TCR-activated or produce IFN-γ. Here, by generating conditional CD1d knockout mice, our goal was to determine which cell types directly activate NKT17 cells during infection. In mice with a CD11c+ cell-specific deletion, there was a 50% reduction of IL-17A-producing NKT17 cells at 14 hours, and a significant increase in bacteria loads at 48 hours. While these mice lack CD1d on several cell types, we focused on CD103+ DCs. If CD103+ DCs are absent, mice are highly susceptible to S. pneumoniae infection, iNKT cells do not expand, and IL-17A is diminished. Using confocal and two-photon microscopy of live tissue sections and flow cytometry we investigated their role as APCs. CD103+ DCs engulf a GFP-expressing S. pneumoniae strain within 2 hours and are primarily located within the interstitial lung tissue, notably where most NKT17 cells localize. We next generated a mouse with a CD103+ DC-specific deletion of CD1d. These mice also showed a 50% reduction of IL-17A-producing NKT17 cells at 14 hours and an increase in bacteria loads at 48 hours. Continuing studies include microscopy of live lung sections to further investigate the direct interaction between CD103+ DCs and iNKT cells during S. pneumoniae infection.
Dual roles of the bacterial siderophore enterobactin by inducing apoptosis in macrophages and promoting survival advantages to Salmonella typhimurium
Authors: Saha Piu , Xiao Xia , Yeoh Beng San , Chen Qiuyan , Katkere Bhuvana , Kirimanjeswara Girish Soorappa , Vijay-Kumar Matam 
Published: January 2023

Abstract
Iron, an essential transition metal ion, is required to facilitate the antimicrobial and redox activity of heme proteins expressed in macrophages (Mφs). In this study, we investigated whether the chelation of iron by enterobactin (Ent; a prototypical bacterial siderophore) can affect the immune responses of macrophages. Herein, we demonstrated that exogenous Ent confers protection to the intracellular pathogen Salmonella enterica serovar Typhimurium from Mφs by promoting M1 to M2 polarization, reducing the expression of pro-inflammatory cytokines and impairing their intracellular killing. Accordingly, WT Salmonella survived better compared to Ent-deficient strain (ΔentB) in Mφs. The addition of apo-Ent (iron-free), but not ferric-Ent, rescued both strains, thus suggesting that the immuno-modulatory effect of Ent could be likely mediated via chelation of Mφs cellular iron. To determine whether the effects of Ent on Mφs could be due to altered cell viability, we next assessed the Mφs for markers of apoptosis via flow cytometry. Interestingly, we observed that apo-Ent, but not ferric-Ent, promoted a time-dependent apoptosis in Mφs, which can be detected as early as 12h after Ent treatment. These findings corroborated with the time-dependent release of lactate dehydrogenase (cytotoxicity marker) from Ent-treated Mφs. Collectively, our findings demonstrate that Ent not only facilitates bacterial iron uptake but also serves to safeguard bacteria against Mφ innate immune responses, which could be mediated, in part, by inducing delayed cellular apoptosis.