bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comprehensive characterization of the mast cell surface proteome
Authors: Shubin Nicholas J , Niino Kerri , Kasprzak Victoria , James Richard , Piliponsky Adrian Martin 
Published: January 2023

Abstract
Mast cells are known for their involvement in many pathological conditions, suggesting that mast cell responsiveness to various stimuli requires the expression of a rich cell surface protein repertoire. Mast cell surface proteomes have not been comprehensively characterized; therefore, our aim in this study was to identify unknown mast cell surface proteins to obtain a better understanding of how these cells function in health and disease. For this purpose, we enriched for glycosylated surface proteins expressed in murine bone marrow-derived mast cells (BMCMCs) and we identified the enriched proteins by using mass spectrometry analysis. The use of this approach resulted in the identification of 435 glycoproteins in BMCMCs. Based on the UniProtKB database, we identified 60 plasma membrane proteins among the enriched glycoproteins, including cell surface receptors, ligands, proteases, and adhesion molecules. The usefulness of our approach was validated by the presence of surface molecules known to be expressed on BMCMCs, including c-Kit, IL2rα, FcɛrIα, Csf2rb, and Tgfβr1. Notably, we identified 22 proteins that have not been previously reported to be expressed in BMCMCs. The presence of these proteins in mast cells was validated by cross-referencing them against the ImmGen database and/or qPCR and western blot analyses. Importantly, variations or deletions in genes that encode for these surface proteins are associated with disorders in which mast cells play an active role, such as infections, asthma, and sepsis. Taken together, our study suggests that we can identify surface molecules that may influence mast cell biology in health and disease.
Profile of Dendritic Cells Surface Biomarkers in Nasal and Oral Mucosa of Human Subjects Reactive to Grass Pollen and House Dust Mites
Authors: Rivera-Mariani Felix Emanuel , Srour Hayat , Fomenko Ruslan , Bellinger Shandra , Stateman Ariel , Baguley Joshua 
Published: January 2023

Abstract
Dendritic cells (DCs) play key roles in human allergic pathophysiology at different anatomic sites, such as the respiratory tract. To identify relationships between DCs subpopulations in the upper respiratory tract and atopic status, a public-available dataset (http://doi.org/10.5281/zenodo.50228) on nasal and oral DCs cell-surface biomarkers expression following allergen challenge (through skin prick test) was re-analyzed, with regression and principal component analysis (PCA), to identify profiles of DC cell-surface biomarkers based on the allergen reactivity. The dataset included immunohistochemical measures on CD207 (langerin), CD1a, BDCA-1 to -4 nasal and oral epithelium and lamina propia, as well as demographic information (age, gender), history of respiratory disease, and allergen skin provocation (grass pollen or house dust mite). Subjects with reactivity to grass pollen had different DCs cell-surface biomarker profiles than subjects reactive to house dust mite in both nasal and oral mucosal epitheium and lamina propia. Except for BDCA-3 and 4, reactivity to grass pollen yielded higher expression of DC cell-surface biomarkers in nasal epithelium. In nasal lamina propia, subject reactive to grass pollen had higher expression of BDCA-1 and -2. In the oral epithelium and lamina propia, subjects reactive to house dust mite had higher expression to only CD1a. Graphical evaluation with PCA of both nasal and oral mucosa of DC cell-surface biomarkers yielded principal components explaining three clusters in oral mucosa; two in nasal mucosa. These findings suggest that different allergen activate differently specific innate immune cells, including different DCs subpopulations at different anatomical sites.
Lymph node conduits transport virions for rapid T cell activation
Authors: Reynoso Glennys V , Weisberg Andrea S. , Shannon John P. , McManus Daniel T , Shores Lucas , Americo Jeffrey L , Stan Radu V , Yewdell Jonathan W. , Hickman Heather D. 
Published: January 2023

Abstract
Despite intense interest in antiviral T cell priming, the routes of virion movement in lymph nodes (LNs) are imperfectly understood. Current models fail to explain how virus-infected cells rapidly appear within the LN interior after viral infection. To better understand virion trafficking in the LN, we determined virion and infected-cell locations after pox- and zika-virus administration. Surprisingly, many rapidly infected cells in the LN interior were adjacent to LN conduits. Using confocal and electron microscopy, we clearly visualized virions within conduits. Functionally, CD8+ T cells rapidly and preferentially associated with vaccinia-infected cells deeper in the LN, leading to T cell activation in the LN interior. These results reveal that, contrary to dogma, even large virions can flow through lymph node conduits to infect T cell zone DCs that prime CD8 + T cells.
Anti-viral CD8+ T cell responses are impaired by endogenous n-3 polyunsaturated fatty acids
Authors: Seo Young-Jin , Kim Seyoung , Cho Yong-Bin , Kang Kyung-Won , Lee Sang-Myeong 
Published: January 2023

Abstract
Omega-3 (n-3) polyunsaturated fatty acids (PUFAs) have been known to exert anti-inflammatory effects on various inflammatory diseases. However, its role in CD8+ T cell responses against an acute viral infection has not been well elucidated yet. To determine the role of n-3 PUFAs in anti-viral CD8+ T cell responses, we used FAT-1 transgenic mice that are able to convert n-6 PUFAs to n-3 PUFAs. The FAT-1 mice or mice orally administrated with n-3 PFUAs exhibited a significant reduction of anti-viral CD8+ T cell responses against an acute strain of lymphocytic choriomeningitis virus (LCMV). When LCMV-specific P14 CD8+ T cells carrying fat-1 gene (P14/FAT-1) were adoptively transferred into mice that were subsequently infected with LCMV, expansion of the cells was substantially suppressed. Similarly, when P14/FAT-1 cells were stimulated with LCMV gp33 peptide in vitro, their expansion was significantly reduced as compared to P14 cells. Collectively, our results indicate that n-3 PUFAs attenuate anti-viral CD8+ T cell responses against acute viral infection.
TSC1-mTOR-controlled metabolic-epigenetic crosstalk underlies DC control of CD8+ T cell homeostasis
Authors: Xiao Hui , Shi Lei , Chen Xia , Zang Aiping , Li Tiantian , Yang Jianbo 
Published: January 2023

Abstract
Dendritic cells (DCs) play pivotal roles in T cell homeostasis and activation, and metabolic programing has been linked to DC development and function recently. However, the metabolic underpinnings corresponding to distinct DC functions remain largely unresolved. Here we demonstrate a special metabolic-epigenetic coupling mechanism orchestrated by TSC1-mTOR for the homeostatic DC function. Specific ablation of TSC1 in the DC compartment (TSC1DC-KO) largely preserved DC development, but led to pronounced reduction in naïve and memory-phenotype CD8+ T cells, a defect fully rescued by the simultaneous deletion of mTOR or Raptor in DCs. Moreover, TSC1DC-KO mice were unable to launch efficient antigen-specific CD8+ T effector responses required for containing Listeria monocytogenes and B16 melanomas. Mechanistically, our data suggest that steady-state DCs tune down de novo fatty acid synthesis through TSC1 and divert acetyl-CoA for histone acetylation. Correspondingly, TSC1-deficiency elevated ACC1 expression and fatty acid synthesis in the steady-state DCs, leading to impaired epigenetic imprinting on selective genes such as MHC-I and IL-7. Remarkably, tempering ACC1 activity was able to avert cytosolic acetyl-CoA for histone acetylation and restoration of the gene expression program compromised by TSC1-deficiency. Taken together, our results uncover a crucial role for TSC1-mTOR in metabolic programing the homeostatic DCs for T cell homeostasis, and implicate metabolic-coupled epigenetic imprinting as a paradigm for DC specification.
Commensal polysaccharide-induced T cell communication and peripheral immune regulation
Authors: Zhou Julie Y , Jones Mark B , Alvarez Carlos A , Johnson Jenny L , Cobb Brian A 
Published: January 2023

Abstract
The increasing prevalence of inflammatory and autoimmune diseases across the world has been attributed in part to a shift in microbial exposure both in the exogenous environment and the endogenous microbiome. Here we identify and adopt a strategy used by a common commensal gut microbe, Bacteroides fragilis, to suppress inappropriate immune activation. Bacteroides fragilis produces a protective capsule containing polysaccharide A (PSA), which has been shown to suppress a host of inflammatory diseases in an IL-10-dependent manner. We have discovered that PSA is endocytosed, processed, and presented by APCs through MHCII and αβTCR engagement, and clonally expands CD4+FoxP3−CD25−CD45Rblo cells of an effector memory subset (RbloTEM). RbloTEM are capable of suppressing peripheral inflammation in way dependent on IL-10, however, they themselves do not produce the IL-10 necessary for suppression of disease. We show here that RbloTEM cells communicate directly with FoxP3+ Regulatory T cells, resulting in enhanced IL-10 production. Using novel FoxP3RFP/IL-10GFP dual reporter mice, we further demonstrate that this communication induces synergistic IL-10 expression specifically in Tregs, enhances Treg proliferation, and drives Treg-mediated immune inhibition by preferentially favoring the expansion of IL-10+ Tregs. Through promoting a robust and rapid Treg response, we show that the cytokine signature of PSA-responding RbloTEM cells and coordinating FoxP3+ Tregs can suppress mouse models of inflammatory disease, including HDM-induced asthma and EAE.
Visualising interaction of monocytes with red blood cells (RBC) sensitised with “clinically significant” antibodies
Authors: Dean Melinda M , Ji Ju , Gu YuanTong , Sauret Emilie , Flower Robert L 
Published: January 2023

Abstract

Background
Anti-RBC antibodies in plasma are regarded as clinically significant when they promote clearance of transfused RBC by monocytes/macrophages. To date, processes involved in the attachment or phagocytosis of sensitised RBC by monocytes have been investigated using light or fluorescent microscopy. We used scanning electron microscopy (SEM) to facilitate a closer examination of the interaction between monocytes and RBCs sensitised with antibodies of specificities classified as clinically significant.

Methods
Peripheral blood mononuclear cells (PBMC) were seeded onto glass coverslips, and non-adhered cells were removed after 1hr. Antigen-positive RBCs were opsonised with clinically significant antibodies (anti-A, anti-B, anti-D and anti-K) before addition to adhered monocytes. Anti-M was used as an example of an antibody regarded as non-clinically significant. Coverslips were then prepared for SEM which included glutaraldehyde fixation followed by post-fixation with osmium tetroxide. Cells were further dehydrated with ethanol and hexamethyldisilazane before coated with gold nanoparticles

Results
We observed the interaction between monocytes and sensitised RBCs using SEM. Clinically significant antibodies induced monocyte activation, evidenced by multiple morphological changes including cell elongation, extension of membrane ruffles and the projection of pseudopodia to capture sensitised RBCs.

Conclusions
Detailed characterisation of interaction between monocytes and sensitised RBCs provides insight into the processes associated with RBC removal. The model provides a basis for in-depth analysis of the mechanisms associated with antibody-induced RBC destruction in vivo.
CD31 acts as a checkpoint molecule and is modulated by FcγR-mediated signaling in monocytes
Authors: Butchar Jonathan P , Merchand-Reyes Giovanna , Robledo-Avila Frank , Buteyn Nathaniel , Gautam Shalini , Santhanam Ramasamy , Fatehchand Kavin , Partida-Sanchez Santiago , Tridandapani Susheela 
Published: January 2023

Abstract
Monocytes and macrophages express Fc-gamma Receptors (FcγR) that engage IgG immune complexes such as antibody-opsonized pathogens or cancer cells to destroy them by various mechanisms including phagocytosis. FcγR-mediated phagocytosis is regulated by the concerted actions of activating FcγR and inhibitory receptors such as FcγRIIb and SIRPα. Here, we report that another ITIM-containing receptor, Platelet Endothelial Adhesion Molecule-1 (PECAM1/CD31), regulates FcγR function, and is itself regulated by FcγR activation. First, qRT-PCR and flow cytometry analyses revealed that monocyte FcγR activation leads to significant downregulation of CD31 expression, both at the message level and at surface expression. Interestingly, the kinetics of downregulation between the two varied, with surface expression reducing earlier than the message. Experiments to analyze the mechanism behind this discrepancy revealed that loss of surface expression was due to internalization, which depended predominantly on the PI3 Kinase pathway. Finally, functional analyses showed that downregulation of CD31 expression in monocytes by siRNA enhanced FcγR-mediated phagocytic ability. Together, these results suggest that CD31 functions as a checkpoint receptor that could be targeted to enhance FcγR functions in antibody-mediated therapies.
Changes in Myeloid-Derived Suppressor Cell function during neonatal sepsis
Authors: Vance Jordan , Rawson Travis , Witt Michelle R. , Seman Brittany G. , Bradford Shelby , Robinson Cory M. 
Published: January 2023

Abstract
Infants are more susceptible to infectious disease due to immaturity of the immune system. Our laboratory has shown that the immune suppressive cytokine, interleukin (IL)-27, is elevated in neonates. We and others have also shown that myeloid-derived suppressor cells (MDSCs) are more abundant in neonates, and these cells are a source of IL-27 in neonates. MDSCs suppress immunity with additional effector functions such as nitric oxide synthase (NOS2), arginase-1 (ARG1), and production of oxidative radicals to. We hypothesized that MDSCs gain enhanced immune suppressive function during neonatal sepsis. We utilized a murine model of neonatal Escherichia coli-induced sepsis that that demonstrates reduced weight gain in infected animals compared with controls, and high levels of bacteria in the blood and peripheral tissues. MDSCs were isolated from control or infected pups and the expression of effector function genes evaluated. Gene expression analysis revealed that NOS2, ARG1, and IL-27 gene expression was increased in MDSCs from infected mice. To understand whether or not these changes manifest at the protein/functional level and to further understand if they could be replicated in vitro, MDSCs isolated from control mice were cultured with E. coli. A similar pattern of changes in expression was observed with NOS2, ARG1, and IL-27 genes in vitro when the MDSCs were cultured with bacteria. This was followed by increased arginase activity, elevated nitrite levels, and secreted IL-27. These results suggest that gram-negative bacteria can directly act on MDSCs to enhance effector function associated with immune suppression. They also support further study of MDSC activity in regulating host control of infection early in life.
Retinoic acid-induced expression of CD103 in human DCs is dependent on p38 MAPK signaling and NFAT1c
Authors: Roe Mandi M , Swain Steve , Bimczok Diane 
Published: January 2023

Abstract
Mucosal CD103+ (αE integrin+) DCs are critically involved in driving a T regulatory response and enabling mucosal homeostasis. However, the regulation of CD103 expression in human DCs is not fully understood. We previously found that retinoic acid (RA) induces CD103 expression in human monocyte-derived DCs (MoDCs). Here, we investigated the molecular mechanisms leading to RA-induced CD103 expression on human DCs. We used a phospho-protein array to screen RA-treated MoDCs and found multiple potential targets. We focused on p38 MAPK based on previous studies that showed reduced CD103+ DCs in p38 MAPK KO mice. Western blots confirmed increased phosphorylation of p38 MAPK in RA-treated MoDCs. To determine whether p38 MAPK signaling influences CD103 expression, we next used an inhibitor of p38 MAPK and evaluated CD103 expression on MoDCs. The p38 MAPK inhibitor SB202190 significantly decreased expression of CD103 protein and mRNA in RA treated, but not in control MoDCs (p=0.037 and p=0.047 respectively). The transcription factor NFAT1c is a downstream target of p38 MAPK that has been shown to bind to the promotor region of CD103 in T cells. To determine whether NFAT1c regulates RA-induced CD103 expression, we next exposed RA-treated MoDCs to an NFAT1c peptide inhibitor. We demonstrated a significant dose-dependent decrease in CD103 protein expression of RA-treated MoDCs with increasing concentrations of the NFAT1c inhibitor (p=0.03). These data indicate that RA-induced expression of CD103 is dependent on p38 MAPK signaling and NFAT1c in human DCs and give insight into molecular mechanisms of DC signaling.
HMGB1-induced ILC2s activate dendritic cells by producing IL-9 in mice asthma
Authors: Xu Huaxi , Wan Jie , Su Zhaoliang , Wang Shengjun 
Published: January 2023

Abstract
High-mobility group box-1 (HMGB1), a unique and important pathogenic protein, has been implicated as a pro-inflammatory cytokine involved in the pathogenesis of various inflammatory and autoimmune diseases, and CD4 T-cell differentiation and modulation. It has been reported that HMGB1 can promote Th2 response, aggravating the severity of asthma in mice, thus ILC2s contribute to Th2 polarization and maintain this immune imbalance. However, little is known about possible role of HMGB1 on regulation of ILC2s and on the activity of dendritic cells (DCs) in asthma. In this study, we detected the IL-9 expression level in HMGB1-stimulated ILC2s in vitro or asthmatic mice, analyzed the effect of IL-9 derived from ILC2s on DCs function and their role in the development of asthma. Our results showed that HMGB1 could activate ILC2s and promoted IL-9 secretion, and A-box of HMGB1 eliminated this phenomenon. In asthma mice model, A-box of HMGB1 relieved asthma severity. Bone marrow-derived dendritic cells (BMDCs ) could be activated in ILC2s and BMDCs co-culture system, but this effect was inhibited when anti-IL-9 was added in vitro. In addition, IL-9-activated BMDCs co-cultured with naïve T cells promoted the Th2 cells differentiation. The Jak1/stat3 and NF-κB pathway contributed to BMDCs activation involved by IL-9. In asthma mice model, injection of anti-CD90.2 and anti-IL-9 could relieve asthma severity, but transferred exogenous IL-9 made asthma worse. It indicated that increased HMGB1 in the lung of asthmatic mice activated ILC2s to secrete IL-9, which regulated the activity of DC involved in promoting Th2 cells differentiation and finally aggravated asthma.
STEM knowledge translational outreach efforts – connecting with Immunology in the classroom for Pacific Intermediate students
Authors: Sika-Paotonu Dianne 
Published: January 2023

Abstract
The appropriate contextualisation of complex scientific content for young students can be a challenge. Supporting improved interest and engagement with STEM related content by all students in the classroom at the same time, can also be difficult. The purpose of this work was to use an Immunological in-class teaching focus to improve interest in STEM related teaching content for underrepresented student groups at Intermediate school level. A STEM knowledge translational programme organised by an external organisation, included as part of their programme an outreach activity contribution made by a female Pacific Biomedical Scientist for Pacific students aged 10–12 years (Year 7 & 8). Science related learning that was context specific was a primary focus for the Pacific students. Clinical research work components relating to the autoimmune condition Acute Rheumatic Fever (ARF) were presented to the Pacific College students as high rates of ARF and RHD are found in Māori and Pacific Peoples in NZ and the Pacific Region. Enthusiastic in-class interactions between the Biomedical Scientist and the Pacific students were observed. It was noted that the students present were highly motivated to ask questions relating to the Immunologically based content that had been presented. Verbal and written feedback indicated the students (and teachers) enjoyed this particular session and were challenged to consider undertaking STEM related courses in future. This work demonstrated the importance of promoting STEM related teaching content that is of clinical relevance and presented appropriately for Pacific students. The value of using of role modelling in this context was also recognised.
An intervention to increase accuracy and understanding in undergraduate micropipette use
Authors: Avery Lyndsay , Dukes April , Warner Marcie 
Published: January 2023

Abstract
Micropipetting is an essential skill for the budding immunologist and is widely used across biological and physical science disciplines, yet few undergraduates are adequately introduced to this skill. The mentors of undergraduates seeking research experience point to micropipette inexperience as a common barrier to success. As part of our SEA-PHAGES authentic research laboratory, we designed a hands-on intervention to give thorough training in micropipetting to first-year undergraduates. The intervention included assignment of a pre-class video on micropipetting, an activity to orient students to the micropipette parts and functions, and a TA-led demonstration of proper micropipette use and volume adjustment. Success of the intervention was analyzed using data collected from a practical test following the lab activity. In addition, we compared the students’ end-of-semester experimental success to that of students from previous semesters lacking the micropipetting activity, using archived electronic laboratory notebooks. We found that students progressed through experiments faster and with more success after experiencing the micropipette intervention. Our analysis showed that some aspects of proper micropipette use need to be emphasized, such as proper depth of tip entry and the correct tactile discrimination of the first/second stop on the micropipette. In addition, we found that students have misconceptions and gaps in knowledge about the metric system that are an additional barrier to proper micropipette use. This project presents a successful platform for teaching micropipette technique, and demonstrates that this leads to increased likelihood of success for the students in this real-world laboratory setting.