Summary Adenoviral infections represent a major cause of morbidity and mortality following haematopoietic stem cell transplantation. Current anti‐viral agents are virostatic and it is evident that elimination of adenovirus (ADV) infection is only achieved by recovery of cellular immunity. Using an interferon‐gamma (IFN‐ γ ) secretion and capture assay to isolate ADV‐specific T cells, followed by a 2 week expansion and restimulation protocol, we generated ADV T cells that may be used for cellular immunotherapy. In contrast to virus‐specific T cells for cytomegalovirus or Epstein‐Barr virus, the ADV response was dominated by CD4 + T cells and the majority of captured cells exhibited an effector/memory immunophenotype. Highly specific antigen responses were demonstrated by intracellular IFN‐ γ expression and cytotoxicity assays when the expanded cells underwent restimulation with ADV‐pulsed target cells. Although T cells were initially generated in response to ADV species C, the expanded populations also showed strong activity against ADV species B, suggesting cross‐reactivity across ADV species; a finding that has important clinical consequences in the paediatric setting, where the majority of infections are caused by ADV type B and C. The protocols can be readily translated to generate ADV‐specific T cells suitable for clinical use and offer an effective immunotherapeutic strategy to control ADV infection.
Science, Technology, Engineering, and Mathematics (STEM) students are typically taught content delivered didactically and closely aligned with the laboratory demonstration of concepts, which facilitates the development of experimental skills. Due to the volume of content delivered across multiple courses, student cognitive abilities can be affected, leading to lower student performance. In physiology and related biological sciences, educators have turned to delivering content using virtual teaching technologies, including virtual and augmented reality, simulations, and other immersive platforms. At the University of South Australia, Articulate Storyline, Unity-based simulations, and immersive software platforms have been implemented across the entire Laboratory Medicine program to assist students in learning lecture and laboratory content. The impact of these individual interventions is outlined in this manuscript. In addition, the final year 2024 cohort is the first group who have used simulations throughout their degree. Evidence of the benefits and impact of the scaffolded implementation of simulations and immersive software was obtained through a Likert-style questionnaire. The deployment of simulations and immersive software across the degree has significantly enhanced student learning, and engagement with the content effectively bridging the gap between understanding lecture and laboratory content of students in the Laboratory Medicine program. We suggest that a similar approach could readily be embedded within individual courses as well as across science programs to provide the same benefits to student learning.
The gradual shift to online modes of learning in higher education institutions over the past 2 decades accelerated drastically on a global scale between 2020 and 2022. Students and educators, who have initially grappled with the shift, have now become accustomed to online teaching; however, there are concerns about the quality of learning that has resulted. To enable a sustainable and effective online pedagogy, educators may need to learn about fostering higher-order thinking skills, which can be challenging even for experienced educators. To conceptualise effective online pedagogy, the community of inquiry (CoI) framework emphasises cognitive presence (CP), which focuses on the higher-order thinking process. The CoI is the most widely researched framework in online pedagogy, yet contemporary CoI literature lacks collective evidence of factors that influence CP. This scoping review of the CoI literature explores the factors that influence the higher-order thinking that is indicative of CP. Inclusion criteria included evidence of CP in online learning contexts and published between January 2000 and March 2022, providing a total of 121 studies. Results suggest that teaching presence, structure of learning activities and student characteristics all influence CP. Implications for practice or policy: Higher education students enrolled in online courses should be taught how to learn effectively in an online mode. Online course educators must embed learning tasks that foster self-regulation and higher-order skills in students. Online course design should include authentic tasks for students to apply new knowledge to real-life scenarios. Educators must be offered ample professional development activities to build their skills in online pedagogy. Institutions should encourage translation of online educational research to practice.
Inducing cell death by the sphingolipid ceramide is a potential anticancer strategy, but the underlying mechanisms remain poorly defined. In this study, triggering an accumulation of ceramide in acute myeloid leukemia (AML) cells by inhibition of sphingosine kinase induced an apoptotic integrated stress response (ISR) through protein kinase R-mediated activation of the master transcription factor ATF4. This effect led to transcription of the BH3-only protein Noxa and degradation of the prosurvival Mcl-1 protein on which AML cells are highly dependent for survival. Targeting this novel ISR pathway, in combination with the Bcl-2 inhibitor venetoclax, synergistically killed primary AML blasts, including those with venetoclax-resistant mutations, as well as immunophenotypic leukemic stem cells, and reduced leukemic engraftment in patient-derived AML xenografts. Collectively, these findings provide mechanistic insight into the anticancer effects of ceramide and preclinical evidence for new approaches to augment Bcl-2 inhibition in the therapy of AML and other cancers with high Mcl-1 dependency.
Immunology is a complex and rapidly evolving discipline. The complex interplay of multiple cells, mediators and immune modulating agents exacerbates the intricacy of the content making it challenging to teach. However, the nexus between immunology trained scientist and educator has led to the development of innovative approaches that have facilitated student learning. Educational research in immunology not only aids in clarifying difficult concepts, but also addresses the need to prepare students for careers that demand critical thinking, laboratory skills and interdisciplinary fluency. In response to the growing trend of educational research within the discipline, we have curated a Special Feature to highlight examples of educational research that shape the future of Immunology Education, equipping students with the tools to succeed both academically and professionally. We present immunology-focused educational insights from the USA, Australia and the UK. These contributions explore: (1) the scaffolding of interdisciplinary knowledge (Bruns et al.1); (2) the use of digital active learning tools to engage students in both theoretical (Webster et al.2) and laboratory (Costabile et al.3) aspects of immunology; and (3) the preparation of postgraduate students through immersive scientific conference experiences (Rutschmann et al.4). When developing an immunology course, it's essential to ensure that the course is structured with appropriate scaffolding to support student learning; this is key when a cohort includes students from diverse degree programs, varying prior knowledge or differing entry requirements due to advanced standing. Bruns et al.1 explored the development of interdisciplinary science competencies in undergraduate immunology students. They assessed how students in 2nd–4th years integrated their basic science knowledge (e.g. protein structure, receptor–ligand interactions) into understanding immunological concepts (e.g. cytokine function). By their 2nd year, students had mastered most basic science concepts, except "levels of protein structure", which was better understood by the 3rd year. Students reported a better grasp of receptor–ligand interactions when taught within the context of immunology. Confidence in basic science knowledge increased through 3rd year, but no significant improvement was observed by the 4th year. The study highlights that students develop an interdisciplinary understanding of immunology early, suggesting the need for continued reinforcement in later courses to further enhance competency. This embodies the principle of reverse scaffolding given that learners will often fail to recall previous content, so continual reinforcement is important to ensure knowledge retention. Webster et al.2 explores the development of an interactive, online learning activity, designed to aid undergraduate student understanding of T-cell development. This topic is challenging due to the amount of content and technical vocabulary used. The authors created an activity where students assume the role of a T-cell progenitor navigating T-cell development through decision-making scenarios. Students significantly enhanced their understanding of T-cell development while also finding the activity enjoyable. This study concludes that this interactive, asynchronous approach may be effective in engaging students in active learning, even when there are challenges such as social anxiety and resistance to didactic/lecture style formats. Learners can be engaged in narrative-based activities since they activate cognitive and emotional processes, facilitating learning in an engaging and memorable manner. Storytelling can make it easier for students to learn and to remember the content whilst gaining critical thinking and problem-solving skills. Costabile et al.3 investigates the effectiveness of a virtual hemocytometer to teach students how to manually count white blood cells and to perform cell density and viability calculations. The correct use of a manual hemocytometer is an essential skill in diagnostic and research settings. This skill forms the basis for many advanced techniques, building technical proficiency, precision, and a deep understanding of cellular processes, integrating knowledge from multiple areas and experimental design. The study first used the online simulation to train students in all aspects of hemocytometer use, followed by a face-to-face laboratory session, where these skills could be applied, enhancing a student's ability to identify, count and calculate cell density and viability of lymphocytes and neutrophils purified from whole blood. Student feedback was overwhelmingly positive, with 92% of students supporting the use of similar simulations in other courses. Both students with and without prior hemocytometer experience showed significant increases in confidence after using the simulation. All students agreed that the simulation was authentic and realistic. Simulations enable unlimited practice, demonstrate issues commonly seen in a laboratory setting, including cell overlap, contamination and cell death. While students appreciated the simulation, they did value the opportunity to learn hands-on skills. This approach offers a safe and flexible alternative for skill development which can be applied across multiple topics. Rutschmann et al.4 examine how best to prepare postgraduate immunology researchers when attending a scientific conference. Conferences are one of the most rigorous and dynamic mediums for engaging deeply with one's discipline. The paper evaluated the Immunology in Practice module, introduced in 2019 as part of the MSc in Immunology program, which enables students to attend the British Society for Immunology Congress. The module, designed to immerse students in the immunology community, had high satisfaction levels across multiple cohorts. The evaluation also identified several challenges and areas for improvement. The transition to online during COVID-19 affected student experiences, even though many students had no prior conference experience to compare. Challenges with an online format included the lack of recorded sessions, which impacted notetaking, while in-person attendance in later cohorts led to improved satisfaction with the variety of learning activities. The paper highlights the importance of a community of practice, noting that conference participation fosters mutual engagement and shared learning. While not explicitly measured, several students commented positively on interactions with attendees and their experiences in navigating the scientific community. These interactions were valuable for students' personal and professional growth. While there were areas to refine – such as assessment structures, group work and conference logistics, the module's strengths in fostering collaboration, exposure to cutting-edge science, and the development of professional identities are clear. In conclusion, this inaugural pedagogy edition highlights diverse and innovative approaches in Immunology Education, emphasizing the importance of scaffolding interdisciplinary knowledge, utilizing digital tools for active learning and immersing students in real-world scientific experiences. These studies demonstrate how foundational concepts in immunology can be taught more effectively by integrating theory with practical application, fostering deeper engagement, and preparing students for their professional futures. Whether through interactive online activities, virtual simulations, or exposure to scientific conferences, this Special Feature underscores the need for diverse and adaptable pedagogy that not only enhance learning but also equip students with the core competencies required to excel in immunology. The authors declare no conflicts of interest. Samy Sakkal: Conceptualization; writing – original draft; writing – review and editing. Maurizio Costabile: Conceptualization; writing – original draft; writing – review and editing.
Type 2 diabetes is associated with elevated levels of DNA damage, in particular micronuclei (MNi) which are formed by acentric chromosome fragments caused by double-stranded DNA breaks (DSBs), or whole chromosomes which fail to segregate during mitosis. We investigated if methylglyoxal (MGO), a reactive dicarbonyl known to be elevated in type 2 diabetes is capable of increasing chromosomal instability and DNA damage as measured by the cytokinesis block micronucleus cytome (CBMNcyt) assay in B-lymphoblastoid WIL2-NS cells and primary peripheral blood lymphocytes (PBL). We also investigated the level of various dicarbonyl stress biomarkers, including extracellular and intracellular MGO, protein and MGO modifications of DNA. WIL2-NS cells exposed to either MGO or a glyoxalase 1 inhibitor showed increases in MNi and nuclear buds, which were associated with an increase in intracellular MGO. DNA damage in the form of MNi and nucleoplasmic bridges were observed in primary PBL exposed to 10 µM MGO, suggesting low concentrations of MGO may be genotoxic. Furthermore, we showed, using fluorescent in situ hybridisation, that the majority of MNi caused by MGO in WIL2-NS cells were caused by whole chromosome loss events, rather than DSBs. Our data suggest that MGO, a reactive metabolite elevated in type 2 diabetes and other pathologies, can affect genomic integrity by impairing chromosome segregation during mitosis.
Chronic granulomatous disease (CGD) is mainly caused by mutations in X-linked CYBB that encodes gp91. We have identified two novel mutations in CYBB resulting in the rare X91(+)-CGD variant, c.1500T>G (p.Asp500Glu) in two male siblings and c.1463C>A (p.Ala488Asp) in an unrelated male. Zymosan and/or PMA (Phorbol 12-myristate 13-acetate)-induced recruitment of p47(phox) and p67(phox) to the membrane fraction was normal for both mutants. Cell-free assays using recombinant wild-type and the mutant proteins revealed that these mutants were not activated by NADPH (nicotinamide adenine dinucleotide phosphate). Interestingly, the Ala488Asp mutant was activated by NADPH in the presence of glutathione. These data suggest that the mutations prevented NADPH from binding to gp91(phox) and the requirement of a negative charge at residue 500 in gp91(phox) for NADPH oxidase assembly, in contrast to a previously described Asp500Gly change. These mutations and the effect of glutathione provide a unique insight into disease pathogenesis and potential therapy in variant X91(+)-CGD.
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