bluuebunny/crossref_metadata_2025_split huggingface dataset:
Autologous intraoperative cell salvage ameliorates suppression of transfusion-related monocyte and dendritic cell inflammatory responses
Authors: Dean Melinda M , Obeysekera Maheshi P , Tran Thu V , Sturgess David , Wyssusek Kerstin , Van Zundert Andrea , Gordon Louisa G , Da Silva D , Punnasseril J , Tung John-Paul , Flower Robert L , Roets Michelle 
Published: January 2023

Abstract
ABO compatible allogeneic blood transfusion (ABT) is associated with transfusion-related immune modulation (TRIM) and poor patient outcomes including perioperative infection and myocardial infarction. Intraoperative cell salvage (ICS) enables collection, processing and reinfusion of autologous blood lost during surgery. We investigated whether immune suppression associated with ABT could be reduced by using ICS. Suitable orthopaedic patients were consented and recruited to the study (n=20, HREC/17/QRBW/685). 10mL anticoagulated blood (EDTA) was collected from the patient (pre-operatively) and from the ICS blood product. An established whole blood culture model was used to assess changes to monocyte and myeloid dendritic cell (mDC) inflammatory responses (intracellular cytokine staining for IL-6, IL-8, IL-10, IL-12, IL-1α, TNF-α, MIP-1α, MIP-1β, MCP-1, IP-10) following exposure of the patient’s pre-operative blood to ABO compatible allogenic blood (Blood Service, Kelvin Grove) or their ICS blood (P&lt;0.05 ANOVA). Exposure to ABT significantly suppressed the inflammatory response of both mDC (IL-10, IL-12, IL-1α, IL-8, TNF-a, MIP-α, MIP-1β) and monocytes (IL-10, IL-6, IL-12, IL-1α, IL-8, IL-10, IL-12, TNF-α and MIP-1α). Compared to ABT, exposure to the patients’ own (ICS) blood improved mDC IL-8, IL-10, IL-12, TNF-α and MIP-1α production and monocyte IL-10 and MIP-1β production. We provide in vitro evidence that ICS may reduce ABT associated adverse outcomes associated with improved inflammatory response and immune competence post-transfusion.
Constitutive TGFβ signaling prevents inflammation-induced Langerhans cell migration
Authors: De La Cruz Diaz Jacinto S , Kaplan Daniel H 
Published: January 2023

Abstract
Keratinocytes (KCs) that form the epidermis create a unique barrier niche for cells of the immune system such as Langerhans cells (LCs) and CD8+ resident memory T cells (TRM). These leukocytes provide host-defense but also initiate and maintain autoimmune skin diseases. Hence, elucidating the mechanisms by which CD8+ TRM and LC maintain epidermal residency is of great therapeutic interest. Both cell types require active TGFβ1 to maintain their steady-state epidermal retention. In the epidermis, inactive LAP-TGFβ1 is exclusively activated into active TGFβ1 by integrins ανβ6 and ανβ8 expressed by KCs. However, the relationship between TGFβ1 and LC migration in response to inflammatory stimuli remains unknown. To determine whether enforced TGFβ1 signaling can overcome inflammation-induce LC migration, we generated huLangerin-CreERT2x TGFβRI-CALxL mice (TGFβR1-CALC) that allow for tamoxifen-induced ligand-independent TGFβRI signaling. Enforced TGFβRI signaling in LC prevented migration in response to UVB, chemical haptens, C. albicans epicutaneous infection as well as dermal injection of TNFα and IL1β. LC from TGFβR1-CALC mice did not egress from the epidermis or enter the draining lymph nodes. Notably, in WT mice, we did not observe reduced KC expression of integrin ανβ6 or ανβ8 and levels of pSMAD2 in LC did not decrease in inflammatory contexts. Thus, although enforced TGFβRI signaling can overcome inflammation-induced migratory signals, but the loss of TGFβRI signaling is not required for LC migration. These data are consistent with a model in which TGFβRI signaling drives a LC-intrinsic retention program under homeostatic conditions that is overcome or inhibited by inflammatory cues.
Bactericidal activity of transendothelial neutrophils based the endothelial IL-1α crosstalk during neutrophil–endothelial interaction
Authors: Li Qiuyue , Liu Xiaoye , He Shangwen , Xu Yuanyuan , Zhang Hui , Hou Silu , Dong Hong 
Published: January 2023

Abstract
Neutrophils across endothelial cells to capture and eliminate bacterial pathogens is served as a vocation to prevent infectious diseases during innate immune. However, the mechanisms behind neutrophil-endothelial interaction on the bacterial elimination remains unclear. In this study, we employed a transwell system to co-culture rat intestinal mucosa microvascular endothelial cells (RIMVECs) and neutrophils to illustrate the killing mechanisms of transendothelial neutrophils. We discovered that endothelial interleukin-1α (IL-1α) promoted the survival of rats under Escherichia coli infection. IL-1α enhanced the bactericidal activity of transendothelial neutrophils, while the release of IL-1α is inhibited by lipopolysaccharide (LPS) derived from E. coli during infectious process. In addition, LPS majority damaged RIMVECs on cell membrane and induced cell necrosis. LPS promoted E. coli to escape from neutrophils in vivo and in vitro. Though the isobaric tags for relative and absolute quantification (iTRAQ) method to identify differentially expressed proteins of RIMVECs, we found that IL-1α modulates RIMVECs occurred on the plasma membrane, endoplasmic reticulum and mitochondrial envelope. There were eleven common proteins to persist regulatory. 2 h treatment of IL-1α triggered the cell adhesion molecules (CAMs) of RIMVECs, while oxidative phosphorylation was required for long times’ incubation of IL-1α (4 and 8 h). Our work highlights the critical role of endothelial IL-1α during neutrophil- endothelial interaction, which offered new targets in the bactericidal activity of neutrophils.
Recruitment of kindlin-3 to plasma membrane through its PH domain precedes high affinity β2 integrin activation and neutrophil arrest
Authors: Wen Lai , Marki Alex , Roy Payel , McArdle Sara , Sun Hao , Fan Zhichao , Gingras Alexandre R. , Ginsberg Mark H , Ley Klaus 
Published: January 2023

Abstract
Leukocytes including neutrophils must adhere to the vascular endothelium in order to respond to injury or infection. Integrin-mediated neutrophil adhesion starts by arrest from rolling. Activation of integrins involves conformational changes from an inactive bent conformation to an extended conformation (E+) with high affinity for ligand binding (H+). The cytoplasmic protein kindlin-3 is required for leukocyte adhesion; mutations of kindlin-3 cause leukocyte adhesion deficiency type III syndrome. However, the molecular mechanism by which kindlin-3 regulates β2 integrin activation is unknown. Here we measured the spatiotemporal dynamics of kindlin-3 and β2 integrin conformation changes during neutrophil and HL-60 cell rolling and arrest under flow. Using high-resolution quantitative dynamic footprinting (qDF) microscopy and kindlin-3-fluorescent protein (FP) fusion proteins, we find that kindlin-3 is recruited to the plasma membrane prior to induction of the H+ β2 integrin conformation. Deletion of kindlin-3 or its lipid-binding pleckstrin homology (PH) domain completely abolished H+ β2 integrin induction. Intravital imaging of bone marrow chimeric mice revealed that EGFP-tagged kindlin-3-expressing neutrophils, but not control kindlin-3-deficient neutrophils, arrested in cremaster venules in vivo in response to CXCL1. Our data show that kindlin-3 is indispensable for the H+ β2 integrin conformation and crucial for integrin-mediated leukocyte adhesion during inflammation.
Limited application of IL-2/CD25 leads to persistent IL-2R signaling that promotes gene expression for Treg expansion
Authors: Moro Alejandro , Ban Yuguang , Zhen Gao , Wang Lily , Yu Aixin , Chen Xi , Malek Thomas R 
Published: January 2023

Abstract
Low-dose IL-2 expands Tregs to suppress autoimmunity. However, IL-2 has a short half-life that necessitates frequent administration to maintain increased Treg levels. To improve low-dose IL-2 therapy, we developed an IL-2-based fusion protein, mouse IL-2/CD25, that substantially increases IL-2 half-life to more effectively boost Tregs that controls diabetes in NOD mice. To provide a molecular underpinning for the Treg response to IL-2/CD25, transcriptional profiling was performed by RNAseq for purified Tregs from C57BL/6 mice that received an equivalent amount of IL-2 or IL-2/CD25. Initially, at 4 hr post-injection, differential gene expression analysis revealed a large overlap in significant genes modulated by IL-2 or IL-2/CD25. In striking contrast, at 16 and 72 hr post-injection, only IL-2/CD25 persistently modulated gene expression, with the largest number of IL-2-dependent genes detected at 72 hrs. Canonical pathway analysis revealed many interrelated pathways that control cell growth and signaling. This gene expression pattern by IL-2/CD25, but not IL-2, was also associated with extended STAT5 activation and expansion of Tregs with molecular and phenotypic changes corresponding to activated Tregs. IL-2/CD25 also enhanced gene expression associated with glycolysis and the mitochondrial electron transport chain. Seahorse analysis confirmed that IL-2/CD25 shifted Tregs to a more energetic metabolic phenotype due to an increase in mitocondrial ATP. Collectively, these findings demonstrate that a single injection of IL-2/CD25 persistently stimulates Tregs to activate gene expression to enhance Treg metabolism to fulfill the bio-energetic and biosynthetic demands of activation and proliferation.
Interleukin 7 regulates naive T cell metabolism to promote motility of T cells
Authors: Cannon Judy L , Byrum Janie Rae , Gridley Rebekah , Mrass Paulus , Valenzuela Miriam , Torres David J , Fricke Matthew , Moses Melanie E 
Published: January 2023

Abstract
Interleukin-7 (IL-7) is a key cytokine that drives the survival and maintenance of naïve T cells in lymph nodes. Naïve T cells continuously move throughout lymph nodes in order to interact with dendritic cells (DCs) to become activated and promote an effective immune response. As IL-7 is highly expressed in lymph nodes, we investigated whether IL-7 might regulate T cell movement potentiating T-DC interaction. Using two photon microscopy, we show IL-7 promotes T cell motility in vitro and in vivo. Downstream of IL-7, JAK3 and STAT5 signaling are important to regulate T cell speed. To understand the mechanism underlying IL-7 effects on T cell movement, we investigated whether IL-7 might change the metabolic profile of naïve T cells. Using the Seahorse analyzer, we show that IL-7 can affect the metabolic profile of naïve T cells, particularly CD8 T cells. The chemokine CCL21 has previously been shown to drive T cell motility. We then asked whether IL-7 and CCL21 might combine to modulate effects on T cell metabolism. We find that CCL21 decreases ATP production and this decrease in ATP production persists even in the presence of IL-7. Our results suggest that IL-7 is a novel regulator of T cell motion in lymph nodes via changes in T cell metabolism, leading efficient T cell responses.
Rapid protocol for in vitro induction of cytokine associated with Th2 and Th17 cells from PBMC
Authors: Soto Hortensia , Elia Jeanne , Yu Cuiling , Yu Daniel , Yu Jie , Ni Jessie 
Published: January 2023

Abstract
T helper cell is a key component of the immune system. They mediate humoral and cellular immune responses against pathogens and other diseases like cancer by inducing and activating effector cells such as B cells, cytotoxic T cells and innate cells. They are also involved in the regulation of immune responses by controlling the persistence and magnitude of the response. Activation of naïve T cell is initiated by interaction with antigen presenting cells. The downstream signal transduction and the cytokine milieu can lead to their proliferation and differentiation into different T cell subtypes. To facilitate the study of Th2 and Th17 cells, we present a rapid protocol to efficiently induce cytokine associated with Th2 and Th17 cells from total PBMC. Canonical anti-CD3 and anti-CD28 clones are used to activate the purified cells. Different sets of bioassay-validated cytokines, including IL-12, IL-4, IL-6, IL-1β, IL-2, and TGF-β1 are used to induce cytokine associated with Th2 and Th17 cells. Hallmark cytokine and transcription factor expression profile of different lineages are evaluated by Flow Cytometry. In summary, we have successfully developed a rapid protocols to induce cytokine expression in vitro associated with Th2 and Th17 using high quality reagents developed in BioLegend.
Undergraduate immunology lab project including opportunities for experimental design, a poster presentation, and the opportunity to partially purify antibodies from hybridoma cells to utilize in a Western blot
Authors: Koenigstein Carrie Temple , Johnson Mills 
Published: January 2023

Abstract
Undergraduate students benefit from lab activities that more closely mirror research conditions. In this 7 week lab project, students maintain the hybridoma cell line MARC 2B7 (ATCC CRL-2509), and design a mini-research project to examine antibody production under a modified growth condition of their choice. Lab groups are responsible for making their own solutions, designing controls, and monitoring and maintaining the cell line over a two week period of time. Antibody levels are measured using a dot blot. For each phase, students are presented with a standard lab protocol accompanied by a list of suggestions and/or modifications so that the students are responsible for writing their own final protocol. For example, the dot blot protocol provided is based on chemiluminescent detection; however, the commercial secondary antibody is HRP conjugated so students are also provided the manufacturer’s information on the secondary antibody and the HRP substrate available in lab. This mirrors more real-world lab experiences, and requires students to think more deeply about the experimental design. Each lab group presents a poster on their mini-project to the class. Subsequently, lab groups use ammonium sulfate precipitation followed by a desalting column to concentrate and partially purify the antibodies from hybridoma supernatants that demonstrated positive results on the dot blots. This product is used as the primary antibody for a Western blot at the end of the experiment arc. In addition to providing a platform to discuss both monoclonal and polyclonal antibodies, and enabling students to practice experimental design, this project requires students to utilize their work from early in the semester in a productive final lab experience.
Learning outcomes of using animated video podcasts as a tool for teaching immunology physiology and pharmacology courses in pharmacy curriculum
Authors: Faridi Hafeez , Garcia Ruslan , Castillo Yaimi , Shahid Mohd , Garg Rahul 
Published: January 2023

Abstract
Pharmacy curriculum demands mastery of multiple disciplines such as physiology, immunology and microbiology, medicinal chemistry and pharmacology. Professional pharmacy programs in the United States and other countries have adopted the integrated curriculum model to fulfill the learning objectives. Intensive learning and constant revision of study material is necessary for the success in the pharmacy program. With modern digital technologies and high-speed internet, last decade has witnessed a paradigm shift in traditional pedagogy in every discipline we teach and study. Popularity of online education has grown exponentially in recent years. Several reputed Universities globally now offer blended as well as complete online courses. Large student population from every disciplines now embark on largely available digital resources such as educational videos. As a part of preliminary studies we have prepared few videos and identified some of the potential benefits of using video podcasts in pharmacy curriculum as easy accessibility of material, short video podcast helps better engage students and avoid boredom, animations enhance learning process and memorization, students can study at the ease of watching these videos on phone. Our results suggests strong likability of educational videos as a learning tool among students in pharmacy curriculum.
Teaching immunology laboratory techniques in a course without a lab component using Journal of Visualized Experiments Immunology and Infection, a peer-reviewed scientific journal published in video format
Authors: Walline Crystal C 
Published: January 2023

Abstract
Teaching an undergraduate immunology course without a lab component has many challenges. Firstly, a complicated subject must be taught in about half the number of contact hours. Chiefly, the opportunity to reinforce concepts learned from the instructional setting (e.g. classroom, online module, or in situ teaching) by performing hands-on experimentation is absent. An assignment was devised which required students to read an article published in the Journal of Visualized Experiments (JoVE) Immunology and Infection and watch the companion video. JoVE publishes peer-reviewed detailed experimental techniques in a visual format. Guidelines for critically analyzing a scientific text were provided and students were asked to do a close reading and post questions to an LMS discussion board, which the instructor used to implement the Just-in-Time Teaching (JiTT) method. The assignment was discussed in class by beginning with a list of unfamiliar acronyms and jargon. This provided a starting point for an interactive discussion of the key immunology concepts (e.g. chemotaxis) which was an excellent alternative to a standard lecture. Next, the class watched the video which was paused periodically to address questions posted to the discussion board. An end of discussion assignment was given for students to compare/contrast protein analysis from traditional SDS-PAGE/western blotting to a single-cell, chip-based western blot. Students critically read the literature and learned standard and cutting-edge experimentation/instrumentation in a course without a separate laboratory component. Overall, the use of a visual journal article from JoVE and JiTT led to an interactive, engaging discussion that received positive feedback from students.
Human subjects research as part of immunology education
Authors: Redmond Sarah , Mohamed Attia 
Published: January 2023

Abstract
Original research has been identified as a high impact practice in undergraduate education – students who participate have been shown to be more likely to be be successful in professional and academic graduate programs. Course-embedded research allows for students to tie laboratory experiences to classroom concepts and allows for many students to benefit from faculty oversight. Undergraduate research in immunology presents challenges because most models of immunology are limited to vertebrate organisms, which require additional regulatory oversight from Institutional Animal Care and Use Committees or Institutional Review Boards. One common solution is to limit student interaction with organisms by using isolated or cultured cells, however this prevents students from gaining a realistic understanding of all the stages of immunological research. Obtaining authorization for student-designed research, conducting the projects, and analyzing and presenting the results within a single semester is a daunting proposal, one which has been further complicated by the full implementation of the new Common Rule for human subjects research. We have developed a model for conducting human subjects research within a single semester of a junior-level undergraduate immunology course. By using a framework protocol, non-invasive sampling, and anonymized presented data we are able to work within IRB/IACUC regulations beyond the classroom exercise level, allowing students to share their findings with the broader campus community, in online portfolios, and at professional conferences. This model has been sustainable for five years at a public primarily undergraduate institution and has been successful under the new common rule regulations.
Contextualisation of Immunology related STEM examples to support Youth interest and engagement
Authors: Sika-Paotonu Dianne 
Published: January 2023

Abstract
Efforts are being made to increase the Pacific health workforce in New Zealand (NZ), and also to support the health workforce composition become more reflective of the NZ population. For those embarking upon health based career pathways, this will require participation in STEM and health related subject matter. More work is needed to support the uptake, interest and engagement by Youth in STEM related content – particularly for Pacific students. The purpose of this work was to use STEM related teaching examples with appropriate contextualisation of Immunology based content to support Youth interest and engagement in science amongst Pacific Youth. In-class teaching examples using Cancer related research findings with an Immunological basis were shared with Pacific Youth within group settings that were culturally appropriate and supported group discussion amongst the Youth. These group interactions were facilitated by a Pacific Biomedical Scientist. Positive in-class interactions were supported by the culturally appropriate setting which contributed in supporting favorable student learning and engagement during the classes. Discussion efforts relating to the Cancer related Immunological aspects within these sessions were welcomed by the students, and well-received overall. This work demonstrated the role and importance of cultural considerations needed for Pacific students in supporting positive learning and engagement experiences relating to STEM associated teaching content in the classroom.
Medical immunology for Gen Z
Authors: Geis Abby L , Brown Teressa 
Published: January 2023

Abstract
While there is much discussion in education literature about the benefits of a flipped classroom, medical educators still typically rely on traditional didactic lectures as a primary teaching modality. In fact, when given a choice of delivery modalities, learners often prefer to engage in the easier and comfortable route to knowledge, including listening to lectures. However, traditional learning through passive techniques such as reading and listening is less effective because it fails to facilitate application and recall of concepts. High order learning and retention is significantly improved when the learning process is challenging and thought-provoking.
We use immunology within a medical school curriculum to compare the effectiveness of and student preferences for two distinct teaching modalities. Data will be collected from 170 first year medical students at our institution. The immunology unit consists of 5 sessions, each session linked to 3–4 exam items. Four sessions consist of prerecorded lectures provided to students prior to class. In class, students complete a short individual quiz with low-level questions, the quiz is reviewed, and the remaining class time is spent doing high-level application problems. The final session is delivered as a didactic, in-class lecture. For that final session, students are provided with the low-level quiz and high-level application problems to do at their own discretion. No aspect of the learning activities is required or graded. Following the unit, students will be anonymously surveyed regarding preferences of content delivery modalities. Effectiveness of delivery modalities will be compared by assessing student performance on linked exam items. The unit concludes in November 2019.