bluuebunny/crossref_metadata_2025_split huggingface dataset:
Virtual graduate immunology and immunotherapy education during COVID-19
Authors: Sterner Rosalie , Hedin Karen E 
Published: February 2023

Abstract
The objective was to ascertain the impact of an online, virtual course in immunology and chimeric antigen receptor (CAR)-T cell therapy on trainee’s knowledge and interest in the field. The course occurred May 4–8, 2020. Students taking the course for graduate school credit received a pre-course and post-course survey. The survey included questions covering background, knowledge/confidence in skills, interests, and perceived ability to learn using an online platform. Students worked in teams to develop a research grant specific aims page. A total of 27 students took the course for graduate level credit and were surveyed, and all 27 (100%) returned the surveys. Students came from variable backgrounds and were at different stages of graduate training. After completion of the course, there was a statistically significant increase in self-perceived knowledge of immunotherapy (mean score of 3.7 +/− 1.0 standard deviation post-course vs 2.8 +/− 1.3 pre-course, on a 5 point Likert scale, P=0.0075), knowledge of the translational process (3.8 +/− 0.9 post-course vs 3.2 +/− 1.3 pre-course, P=0.0082), knowledge of the cancer treatment/immunotherapy patient experience (3.5 +/− 1.2 post-course vs 2.4 +/− 1.3 pre-course, P=0.0003), and confidence in immunotherapy grant writing skills (3.7 +/− 0.8 post-course vs 2.7 +/− 1.2 pre-course, P&lt;0.0001). This course can serve as a model to help encourage training in translational research and enhance knowledge and interest in immunology, immunotherapy, CAR-T cell therapy, and regenerative sciences via an online platform in this era of COVID-19 and potentially beyond.
Old is Gold! Old School is Back to a New School of Learning.
Authors: Fazal Nadeem , Brehm Danielle , Jimenez Julianne 
Published: February 2023

Abstract
COVID-19 has challenged the way students' learn and gain knowledge. The purpose of this research project was to investigate how students adapted to remote learning due to COVID-19. We selected a group of undergraduate Biomedical Science students at the University of Birmingham, England with the intent of understanding their new approaches to learning and whether remote teaching discriminated against the less fortunate. Our google form survey received 40 responses. 27 students said they forgot how to learn, with only 15 obtaining new ways of learning. This demonstrated how the adjustment to online teaching detracted from the time and energy students could put into learning. 75% of students did not enjoy university as much as they did pre COVID-19, and 3 students liked no aspects of online teaching. 22 students learnt worse online, while 10 felt no difference. Choosing when to watch lectures improved some students time management, self motivation and discipline. However, lack of externally-imposed routine brought new challenges as students had no time pressure, making it easier to fall behind. Cancelled laboratory work meant students missed out on vital skills. Our data showed that student satisfaction decreased, traditional learning methods were lost, and new ways of learning were hard to attain when transitioning to online teaching. With the prospect of learning returning to what the student’s consider normal, students fear this will accentuate how they have forgotten to learn in the traditional way, posing another disruption to their studies as they readjust to a post COVID world, whenever it comes. We anticipate that lessons learnt and forgotten in a pandemic-induced remote environment will remain a challenge to current students at large.
Instructor attitudes towards digital assessment in undergraduate immunology courses.
Authors: Pearson Todd F , Brayton Kate A , Twitchell Elizabeth A , Bolster Ruth E 
Published: February 2023

Abstract
To be better able to meet the instructional needs of undergraduate immunology instructors, we sought to understand the attitudes of individuals teaching this course. We conducted a survey, inviting 230 randomly selected instructors to participate, of which 38 completed the survey. Our results highlight that while very few instructors (2/38, 5.26%) currently use digital assessment in their immunology course, a strong majority (26/38, 72.22%) indicated that they would use it if it were available with their text. A similar number of instructors (23/38, 63.89%) expressed interest in adaptive quizzing functionality in a digital assessment platform. Additionally, we explored instructor attitudes about the goal of digital assessment. Despite a wide range of responses, over half of the survey respondents identified terminology/vocabulary (19/38, 52.78%) and reading questions that test student understanding of basic concepts (19/38, 52.78%) as important goals of a digital assessment platform. These results highlight that most instructors see digital assessment, especially in a formative setting, as an important instructional modality that can address student learning challenges.
Cell Health Analysis on Guava Muse Analyzer – A Capstone Study
Authors: Chen Thurston , Ghanimifard Bita , Francia Marielle Joy , Gillis Katherine , Salow Lisa , Dickey David , Husarik Aarti , Madiraju Charitha 
Published: February 2023

Abstract
Guava® Muse® Cell Analyzer is a fluorescence detection instrument that performs flow cytometry analysis in an easy-to-use and simple format. The Muse® Cell Analyzer is loaded with immunological and cell health assays optimized using validated kits from the manufacturer. A student-driven research project that is part of a Research & Scholarship (R&S) course series (previously Capstone) incorporated Guava® Muse® Cell Analyzer-based experimental protocols undertaken in collaboration with Luminex Corp. The specific goal is to perform initial characterization of a myeloma cell line for its inherent ability to proliferate and survive. Three proprietary kits designed for cellular assays were helpful in assessing myeloma cell health on Guava®Muse® Cell Analyzer. Students enrolled into the research project worked under the mentorship of a research faculty, reviewed the scientific literature, charted schematics prior to experimentation with the Muse® CellAnalyzer-based protocols for cell death and cell survival, performed quantitative analysis and evaluation of the data. Training on the Muse® Cell Analyzer instrument during R&S course series helped students understand the miniaturized version of the flow cytometry-based technology, evaluate the three Guava®Muse®Cell Analyzer-based immunological methods for cell health assessment, and integrate the knowledge captured during didactic coursework with experimental evaluation of an immune cell type.
COVID-19 affected “face to face” interaction with the professors. Lessons learned from traditional learning.
Authors: Olk Sandra R , Stempek Ryan , Renteria Sahira , Fazal Nadeem 
Published: February 2023

Abstract
Many university programs have implemented/utilized online teaching platforms for various reasons over the last decade. This has allowed for both students and professors to track progress in the course, as well as providing a method to submit assignments over the traditional paper submission. These are just a few of the reasons that online platforms have gained popularity, though many have been caught off-guard by the sudden full transition from traditional/hybrid learning to online learning. Through the use of surveys, we have aimed to study the effects, both positive and negative, that this transition to online learning has had on university students. Our group has pooled our questions into a larger set of survey questions that have been broken into smaller surveys, so that we can encourage a response rate that allows us to maximize our results without having students lose interest in answering all of our questions. In this current research, we have focused on the major loss in the current pandemic induced learning. Our preliminary data suggests that one of the major things that students feel is that they miss face to face interactions with their professors. Since we looked at a cohort of professional pharmacy school students where experiential education is one of the major aspects of training future pharmacists. The students believe that the practical aspect of learning needs supervision, oversight, mentoring, and physical presence of teachers. Most of the students feel that no matter how realistic the internet world of learning seems, it can not compensate the training and practice required in a professional school of pharmacy. We conclude that nothing can compensate or simulate the loss of profound student-teacher interaction.
The integration of Immunology based Pharmacological elements for Health Sciences education
Authors: Sika-Paotonu Dianne 
Published: February 2023

Abstract
Health Sciences education supports learning across multiple discipline areas. Appropriate integration of different subject aspects for students when using particular conditions as in-class teaching examples is important. This work highlights the use of integrated teaching approaches within Health Sciences education to support learning of Immunology based Pharmacology elements relevant to the drug design context for the prevention of Acute Rheumatic Fever (ARF) and Rheumatic Heart Disease (RHD). ARF is the body’s autoimmune reaction to an untreated Group A Streptococcal (GAS) infection of the throat or skin. If a severe, or repeated ARF episodes occur that are left untreated, permanent cardiac damage can occur known as RHD. In the absence of a GAS vaccines, the most effective management plan for ARF/RHD involves painful monthly Benzathine Penicillin G injections for at least 10 years or more. These BPG injections prevent recurrent GAS infections that may trigger ARF and lead to RHD. Adherence to BPG injection regimens remain problematic, and a new Penicillin for ARF/RHD prevention that is less painful and longer lasting, is urgently needed. Local and international research efforts are well underway to support the generation of a new Penicillin for this purpose. When integrated teaching approaches that included online interactive websites were used to support learning of Immunology based Pharmacology elements relevant to the drug design context for ARF/RHD prevention, students demonstrated better in-class engagement, participation and concept understanding, expressing approval and improved confidence with the teaching content.
A novel subset of follicular helper-like MAIT cells has capacity for B cell help and antibody production in the mucosa.
Authors: Jensen Owen , Trivedi Shubhanshi , Meier Jeremy D , Fairfax Keke , Hale Scott , Leung Daniel T 
Published: February 2023

Abstract
Mucosal-associated invariant T (MAIT) cells are innate-like T lymphocytes that respond to conserved microbial metabolites presented on the evolutionarily conserved MHC class I-related protein, MR1. MAIT cells are enriched in the mucosa and aid in protection against bacterial pathogens via release of inflammatory cytokines and cytotoxic molecules. Recent evidence suggests MAIT cells are capable of providing B cell help in vitro and in vivo. In this study, we used primary human samples and mouse models to uncover the role of MAIT cells in B cell help at the mucosa. Here we describe a novel population of CXCR5+ T follicular helper (Tfh)-like MAIT cells (MAITfh) that are enriched in human mucosal lymphoid tissues. Using in vitro culture assays and fluorescence imaging, we found that MAITfh cells preferentially locate within germinal centers in human tonsils and express the canonical Tfh transcription factor, B-cell lymphoma 6 (BCL-6), co-stimulatory markers, inducible T cell costimulator (ICOS), and programmed death receptor 1 (PD-1), and cytokines, interleukin (IL)-21, IL-10, and BAFF. Furthermore, we investigate the sufficiency of MAIT cells to aid in antibody mediated immunity in vivo using a mouse model of MAIT adoptive transfer followed by mucosal challenge with live Vibrio cholerae. We show that adoptive transfer of MAIT cells into αβ T cell deficient mice promotes B cell differentiation and increased serum V. cholerae-specific IgA and bactericidal responses. Our data demonstrate the capacity of MAIT cells to participate in adaptive immune responses at the mucosa, and suggest MAIT cells may be potential targets to improve mucosal vaccines.
Origin of human uterine natural killer cells
Authors: Porrett Paige , Gonzalez Michael , Garifallou James , Smiler Jacqueline , Mafra Fernanda , Silva Renata Pellegrino Da , Johnston Sarah Addie , Vieyra Mark , Testa Giuliano , Johannesson Liza , O‘Neill Kathleen 
Published: February 2023

Abstract
Uterine NK cells (uNK) are a distinct immune population which influence pregnancy outcome. While murine studies have suggested that tissue-resident uNKs predominate in virgin endometrium, the origin of uNKs in humans is unknown. To define subsets of uterine NKs in humans and determine their developmental origin, we performed single-cell RNA-sequencing on CD56+ uNKs sorted from endometrial biopsies of 4 healthy controls (HCs) (17,146 cells) and 5 uterus transplant (UTx) recipients (30,351 cells). SNP polymorphisms and HLA alleles were used to identify cells of donor (tissue-resident) or recipient (peripheral) origin in UTx recipients. Developmental trajectories were built with Monocle v.3.0. Ten major clusters were identified in HCs, with 48% of endometrial NK cells (eNKs) occupying 3 clusters (eNK1–3) that shared gene signatures with known human decidual NK cell subsets (dNK1–3). Pseudotime analysis revealed that the eNK1–3 clusters arose from proliferating subsets and were less mature than cytotoxic uNKs expressing ZEB2, TBX21, and FCGR3A (CD16). uNKs from UTx recipients were similarly distributed across the clusters, with a trend towards fewer cells in the FCGR3A-expressing mature subset. Notably, recipient-derived uNKs dominated all uNK subsets within four months post-transplant but became less frequent in UTx recipients &gt;1 year after transplant (95% vs. 72%; early vs. late). The proportion of donor-derived to recipient-derived cells remained constant across subsets in all UTx recipients. In contrast to mouse models, our study suggests that human uNKs derive primarily from blood-borne peripheral immigrants and that environmental factors influence uNK differentiation more than genotype or developmental origin.
Spatial Map of Human B Cell and Plasma Cell Compartments
Authors: Liu Ailing , Rothrauff Abigail Elizabeth , Borghesi Lisa A 
Published: February 2023

Abstract
Naive B cells differentiate into antibody secreting plasma cells (PC) and memory B cells (MBC) that together dictate the quality and persistence of the humoral immune response. However, a comprehensive analysis of human B lineage population structure that integrates surface phenotype and intracellular functional status is lacking. One reason for this knowledge gap is the difficulty of obtaining diverse tissues from normal individuals. Here we have performed a comprehensive analysis of human lymphoid tissue from relatively inaccessible sites including SPL, BM and LN to visualize B cell organization across diverse tissues from normal donors. We exploited a novel algorithmic pipeline for unbiased analysis. Further, we are aligning findings across mouse and man. Here we report the identification of select markers that are involved in a shared BM precursor/PC program including CD28, CLEC12A, CCR2, CD7 and CD79b by LEGENDScreening. We have validated these markers by RNA-seq analysis of murine splenic PC and conventional flow cytometry analysis of human gut PC. We hypothesize that long-lived PCs returning to BM extinguish the B cell program and re-engage with the ancestral long-lived program. Overall, our findings constitute the basis for a geographic understanding of human B lineage population structure that will inform studies on the human humoral immune response and the mechanisms of PC longevity.
Easy isolation of F4/80 positive macrophages from mouse tissues
Authors: Antignano Frann , Poon Grace , Liang Alice , Eaves Allen , Louis Sharon , Kokaji Andy 
Published: February 2023

Abstract
Macrophages are important and also have critical roles in the development and homeostasis of tissues and organs. Our understanding of macrophages is constantly evolving and expanding, as they are actively studied in many areas of research, including infectious diseases, wound healing, and tumor immunology. Adding to the complexity of macrophage research, subsets of macrophages can be identified throughout the body with diverse phenotypes and functions. Furthermore, macrophage frequency can be highly variable across different tissues, presenting a challenge to obtain highly pure macrophages. To address these challenges, we have developed a simple method to isolate macrophages by targeting F4/80, a well-established murine macrophage marker. Starting with a single-cell suspension from mouse peritoneal cavity, lung, or spleen, F4/80-positive cells were labeled with an antibody complex that links F4/80-positive cells to magnetic particles, then separated using an EasySep™ magnet. Using this method, F4/80-positive cells were enriched from 37.3 ± 9.3% to 94.4 ± 2.9% (mean ± SD; n = 12) from peritoneal lavage fluids, 26.5 ± 2.7% to 94.3 ± 2.8 % (n = 9) from lungs, and 8.0 ± 2.4% to 88.8 ± 3.4% (n = 18) from spleens. Protocols have been optimized to accommodate a range of sample sizes from the various tissue sources. EasySep™-isolated F4/80-positive macrophages are functional, as demonstrated by their ability to uptake FITC-dextran and secrete inflammatory mediators upon activation. Overall, the EasySep™ Mouse F4/80 Positive Selection Kit enables simple and easy isolation of F4/80-positive macrophages in under 25 minutes, streamlining the experimental workflows for researchers studying macrophage biology.
Androgens regulate sex differences in lung-resident ILC2 functional responses during influenza virus infection
Authors: Kadel Sapana , Turner Sean , Karlik Anna , Miller Reegan , Ainsua-Enrich Erola , Kovats Susan 
Published: February 2023

Abstract
Lung-resident group 2 innate lymphoid cells (ILC2s) produce type 2 cytokines and facilitate tissue repair in response to alarmins such as IL-33 released during respiratory virus infection, but they also may be functionally suppressed by type 1 cytokines. Lung ILC2s express notably high levels of intracellular androgen receptor (AR) compared to ILC1s, T and B cells. To test the hypothesis that females and males show differential ILC2 responses in influenza virus (IAV) infection, we analyzed the numbers and functional responses of lung ILCs in IAV infected mice. On days 3–10 post-infection, lungs of female mice contained greater numbers of ILC2s and ILC1s compared to males. However, the female ILC2s were preferentially suppressed at the peak of infection, with a dampened type 2 program manifest as attenuated proliferation, decreased IL-5 and amphiregulin production, reduced expression of GATA-3 and IL-33R and increased surface IFNGR. IFNG levels in the lung were comparable between sexes at day 7 post-infection, suggesting intrinsic differences in ILC2 responses to interferons. Indeed, naïve female ILC2s showed higher expression of IFNGR and higher phospho-STAT1 levels following stimulation by IFNG. Early life orchiectomy revealed that endogenous androgens decreased ILC2 numbers but protected males from suppression of ILC2s in IAV infection. Furthermore, single cell RNA-sequencing experiments revealed that female ILC2s were enriched in a “suppressed ILC2” cluster while male ILC2s were enriched in a “proliferating ILC2” cluster on day 7 post-infection. Taken together, our data show that AR activity preserves canonical ILC2 function in males during IAV infection and may help to explain known gender differences in immunity to IAV.
Histotripsy ablation stimulates the innate immune system and modulates anti-tumor immunity in pancreatic cancer
Authors: Hendricks Alissa Danielle , Sereno Jaqueline , Gannon Jessica , Zeher Allison , Brock Rebecca M. , Betiel-White Natalie , Simon Alexander , Davalos Rafael V. , Coutermarsh-Ott Sheryl L , Vlaisavljevich Eli , Allen Irving C 
Published: February 2023

Abstract
With an overall 5-year survival rate of less than 9%, pancreatic cancer is a substantial cause of cancer related death in the United States. This high mortality has been associated with the immunosuppressive tumor microenvironment (TME) and the shortage of effective late-stage therapeutic options. Histotripsy is a non-thermal, non-ionizing, non-invasive cancer ablation modality that rapidly lyses targeted cells. To investigate how histotripsy can change the pancreatic TME, a Pan02 cell culture model of pancreatic adenocarcinoma was employed to determine the neoantigen release following histotripsy treatment and other standard ablation modalities. Histotripsy was found to release potential neoantigens at a level similar to other non-thermal ablations and superior to thermal ablation, which paralleled the increased innate immune system activation in vivo. For in vivo studies, subcutaneous Pan02 tumors in mice were treated with histotripsy. To monitor changes in the TME after treatment, rtPCR and flow cytometry were used and compared to untreated animals. Progression-free and overall survival were significantly improved with histotripsy treatment. Within 24 hours of treatment there was increased activation of the innate immune system and by 14 days there were decreased tumor-associated immune cell populations. This work illustrates the feasibility of using histotripsy to treat pancreatic cancer and shift the TME as well as providing mechanistic understanding into the activation of the innate immune system post-treatment. Future studies are needed to establish the mechanism of immunomodulation of the TME and the extent of potential systemic effects.
Effects of glucose concentration in the tumor microenvironment on Non Small Lung Cancer (NSCLC) Progression
Authors: Gähler Alexander , Hohenberger Katja , Tausche Patrick , Chiriac Mircea T. , Rauh Manfred , Trufa Denis I , Geppert Carol-Immanuel , Rieker Ralf J , Sirbu Horia , Finotto Susetta 
Published: February 2023

Abstract
Lung cancer is one of the most common cancer types in the western world, with a high lethality and increasing incidence. One major factor, supporting tumor growth is a high sugar diet. Thus, reducing the sugar intake by different diets might be a way to improve current cancer treatment. These metabolic changes are accompanied by regulation of systemic levels of Insulin and insulin like growth factors. As oppose to normal cells, tumor cells metabolism mainly depends on glycolysis. Thus, reducing diet carbohydrate intake might be a way to improve current immunotherapy by inducing immunological changes caused by the extracellular glucose concentration in tumor microenvironment.
Therefore, we cultured the human lung adenocarcinoma (ADC) cell line A549 with different extracellular glucose concentrations to analyze effects by flow cytometry and qPCR. Moreover, lung tissue samples from patients with non small cell lung cancer (NSCLC), were analyzed by qPCR. In a murine model of lung ADC, tumor cells were injected intravenously and mice were fed with different diets. Lung cells were isolated and analyzed by flow cytometry.
Here we show that, IGF1R expression is upregulated in A549 cells line, depending on increasing extracellular glucose concentration. Moreover, in studies on human cohorts of patients with NSCLC, we found a direct correlation between BMI and IGF1R expression and its downstream signaling. Finally, in a murine model of disease, diet glucose restriction decreased significantly tumor load associated with increased lung infiltrating CD8+ T-cells and tissue-resident memory T-cells.
Thus, this study supports the need to introduce glucose restrictions in the diet of patients with NSCLC in association with current immunotherapy.