bluuebunny/crossref_metadata_2025_split huggingface dataset:
MAIT cell function in response to airway epithelial cells is dysregulated in the context of cigarette smoke and COPD
Authors: Harriff Melanie , Larson Emily , Leiser Margalit , Heisler Chelsea 
Published: January 2023

Abstract
MAIT cells are a subset of innate-like lung-resident CD8+ T cells that are thought to play a role in early immune responses to airway infections. These cells exhibit immediate effector function following recognition of bacterial small molecule metabolites presented on the MHC Class I-like molecule, MR1. The impact of cigarette smoke and the development of COPD on MR1 and MAIT cell function is unknown. To address this, we obtained primary airway epithelial cells from the lungs of human subjects with COPD (n=6) or healthy controls (n=6). These cells were treated or not with cigarette smoke extract (CSE), infected with S. pneumoniae, and then used as antigen presenting cells in an IFN-γ ELISPOT assay with a MAIT cell clone. We found that MAIT cells produced more IFN-γ when incubated with uninfected cells from COPD lungs compared to healthy lungs, suggesting that the development of COPD could result in an increase in antigen-independent MAIT cell responses. In contrast, only treatment with CSE impacted antigen-dependent MAIT cell responses to S. pneumoniae-infected airway epithelial cells. Collectively, the differences in MAIT cells responses could not be explained by differences in bacterial infection or MR1 expression in the airway epithelial cells between groups. Dysfunctional MAIT cell responses in the airways could contribute to COPD-associated inflammation by inappropriately activating pro-inflammatory pathways. Furthermore, reduced early recognition of bacterial infection by MAIT cells in the context of cigarette smoke could contribute to persistent colonization of the lower airways and exacerbations.
Modulated MAPK signaling during mast cell differentiation promotes an impaired inflammatory phenotype
Authors: Hicks Natalie Jade , Crozier Robert W.E. , Den Hartogh Danja J. , MacNeil Lori A. , MacNeil Adam J. 
Published: January 2023

Abstract
Mast cells are innate immune cells derived from hematopoietic stem cells. Mechanisms controlling IL-3-directed mast cell differentiation are incompletely defined. MAPK contributes to IL-3R-mediated signaling, yet specific contributions to maturation of functional mast cells are undetermined. We aimed to evaluate the role of the MAPK signaling nodes—JNK, ERK, and p38—in mast cell differentiation, and explore contributions to epigenetic modifier expression. Bone marrow-derived mast cell (BMMC) cultures were established from C57BL/6 mice and differentiated for 7 weeks in the presence or absence of MAPK inhibitors (JNK, JNK-IN-8; ERK, SCH772984; p38, GW856553X). Following inhibitor withdrawal (5 days), phenotypic and functional characteristics of IgE-sensitized BMMCs were assessed using flow cytometry, ELISA, and β-hexosaminidase release assays. Cells differentiated under JNK inhibition showed a significant impairment in the secretion of IL-6 (p&lt;0.05) and IL-13 (p&lt;0.05) and a non-significant impairment in TNF and CCL1 24 hrs post allergen challenge. Degranulation was also significantly decreased (by 25%, p&lt;0.05) in JNK-inhibitor-cultured BMMCs. Expression of c-kit and level of sensitization by IgE was unaffected by impaired JNK signaling. Treatment with p38 and ERK inhibitors did not affect receptor expression, cytokine secretion, or degranulation. Interrogation of MAPK-dependent modifiers of histone acetylation by qPCR and western blot showed that JNK-mediated contributions to mast cell functional responses were independent of HDACs and HATs examined. These results position JNK as a critical contributor to normal mast cell differentiation and potential target in modulating mast cell phenotype.
Altered natural killer cell responses linked to allergen sensitization in patients with atopic dermatitis
Authors: Ohayon David E , DeVore Stanley B. , Biagini Myers Jocelyn M. , Kroner John , Stevens Mariana , Waggoner Stephen N , Hershey Gurjit K Khurana 
Published: January 2023

Abstract
Atopic dermatitis (AD) is a common chronic inflammatory disease associated with skin barrier dysfunction. AD often precedes the development of atopic co-morbidities, including food allergy, asthma and allergic rhinitis. The contributions of natural killer (NK) cells to pathogenesis of AD and progression to allergic disease are unclear. Peripheral blood mononuclear cells (PBMCs) were isolated from blood samples collected from n=82 children participating in the Mechanisms of Progression of Atopic Dermatitis to Asthma (MPAACH) study, a cohort of toddlers with AD. The children underwent skin prick testing to 11 aeroallergens and 6 foods, and wheezing was defined as one or more episodes of wheezing in the past 12 months. We used flow cytometry to evaluate the phenotype of NK cells in patient PBMC from MPAACH subjects. Our analyses revealed that CD56brightNK cells derived from sensitized children expressed significantly less NKG2D (73.8 ± 3.0 % versus 82.3 ± 2.7 %, respectively, p=0.020) and more TIM-3 (440 ± 462 versus 353 ± 391 median fluorescence intensity, respectively, p=0.085) compared to non-sensitized children. Increased expression of TIM-3 on CD56neg NK cells was also associated with wheezing (47.5 ± 0.51 % versus 19.2 ± 0.40 %, p=0.020). Collectively, these observations suggest that changes within the NK-cell repertoire may promote progressive development of AD and atopic co-morbidities in early life.
IFN-γ synergistically activates IL23A gene in LPS-stimulated macrophages
Authors: You Chaelin , Kang Kyuho 
Published: January 2023

Abstract
The cytokine IL-23 is expressed by activated myeloid cells and contributes to the development of Th17-mediated immunity. Dysregulation of IL-23 is associated with chronic inflammatory diseases, but the molecular mechanisms underlying the regulation of IL23A gene in human macrophages, are largely unknown. Unlike murine macrophages, TLR4 signaling alone did not strongly induce expression of IL23A gene but was synergistically activated by IFN-γ priming. To identify human macrophage-specific enhancers in the IL23 gene locus, we used an integrated epigenomic approach including ATAC-seq and ChIP-seq. We found a putative enhancer which is located about 2 kb upstream of the IL23A transcription start site and binds macrophage-lineage-determining factors, such as PU.1 and C/EBP. In this putative IL-23 enhancer, histone acetylation and chromatin accessibility are synergistically increased by LPS stimulation in IFN-γ-primed macrophages. IFN-γ induced coordinate occupancy of STAT1 and IRF1 at IL23A promoter and enhancer. The expression of eRNA and the recruitment of chromatin looping-associated factors cohesin and CDK8-mediator kinase at IL23A enhancer and promoter were increased by IFN-γ priming. The functional relevance of IL-23 enhancer will be tested by eRNA-targeted knockdown and enhancer deletion experiments. These results identify the critical enhancer region in IFN-γ-mediated synergistic activation of TLR4-inducible IL-23 production in human macrophages.
Epigenetic control of the protocadherin-gamma locus provides a potential source of cell surface codes in B cell differentiation
Authors: McDermott Nicole L , Saadi Amel , Cocco Mario , Care Matthew , de Tute Ruth , Owen Roger , Doody Gina , Tooze Reuben 
Published: January 2023

Abstract
Cellular adhesion plays an important role in determining homing and retention of cells to specific locations. Antibody-secreting plasma cells (PCs) depend on access to niche microenvironments for survival, but the factors that control the adhesion behaviour of different PCs are incompletely understood. We noted that cadherin superfamily protocadherin (PCDH) genes showed a dynamic pattern of regulation during differentiation of human PCs.
Clustered PCDHs are split into 3 gene loci (α, β and γ). Choice of variable first exons, which encode the extracellular domain is determined by selective promoter usage. This is controlled by CTCF and enhancer looping. In neurons, PCDH expression generates single cell identity codes important for synapse formation.
Tracking differentiation from B cell to PC stage, ATAC-seq demonstrated selective accessibility over the PCDHG but not PCDHA or PCDHB loci. ChIP-seq for CTCF, H3K4me3 and H3K27ac revealed active promoter marks and CTCF interactions within the PCDHG locus. Flow cytometry confirmed heterogenous expression of PCDHG family members during PC differentiation in line with mRNA patterns. Consistent with post-translational regulation, PCDHG surface expression was additionally increased by ADAM protease inhibition.
We conclude that the B cell lineage shows selective epigenetic accessibility at the PCDHG locus and expression of PCDHG family members upon differentiation to the PC state. This is coupled with patterns of CTCF occupancy implicated in regulating surface identity codes. We conclude that epigenetic control of the PCDHG locus provides a potential source of cell surface codes amongst differentiating PCs.
The role of microRNA-142 in B cell activation and effector functions
Authors: Graham Natalie Marie , Wang Wei-Le , Reyes Estefany Y. , Boldin Mark P. 
Published: January 2023

Abstract
Control of gene expression by microRNA (miRNA) has recently emerged as a critical mechanism that regulates B cell activation and function. However, the role of specific miRNAs in this process is unclear. Using germline knockout (KO) mice, we have previously shown that miR-142 ablation impairs humoral immunity despite significant expansion of the B cell compartment, suggesting that miR-142 is critical for B cell effector function. We found that mice lacking miR-142 cannot generate specific antibody responses upon antigen challenge and display impaired germinal center (GC) formation and plasma cell differentiation. To more precisely dissect the role of miR-142 in B cell effector responses, we developed an activated B cell-specific miR-142 KO mouse by breeding conditional miR-142 KO (miR-142fl/fl) mice with mice expressing Cre recombinase driven by activation-induced cytosine deaminase (AIDCre). AID is required for class switch recombination and somatic hypermutation, and is induced in B cells upon antigen encounter. To label activated B cells in miR-142fl/flAIDCre mice, we bred them to Ai6 mice that carry a loxP-flanked STOP cassette, allowing for the expression of fluorescent ZsGreen1 reporter protein in the presence of Cre recombinase. FACS analysis of Peyer’s patches from naive miR-142fl/flAIDCreZsgreen+/− mice revealed a significant decrease in GC B cells, while the number of ZsGreen1+ activated B cells increased, suggesting an aberrant B cell activation and GC response. We will discuss at the meeting the role of miR-142 in promoting humoral responses to antigen by presenting the findings from our ongoing studies of B cell differentiation and effector function in immunized miR-142fl/flAIDCreZsgreen+/− mice.
IL-2 signals promote acute and selective apoptosis of B cells in Peyer’s Patches
Authors: McKinstry Karl Kai , Singh Ayushi , Dhume Kunal 
Published: January 2023

Abstract
Interleukin-2 (IL-2) is a pleiotropic cytokine demonstrated to be therapeutically effective in treating cancer, but its clinical use can drive severe side effects including gastrointestinal toxicity. Similar symptoms are observed in inflammatory diseases such as Crohn’s Disease. Mounting evidence indicates roles for IL-2 in Crohn’s Disease, but the underlying mechanisms are unknown. Most murine studies aimed at determining IL-2’s in vivo actions on immune cells in the steady state have focused on secondary lymphoid organs. Here, we systemically administered IL-2:anti-IL-2 antibody (clone S4B6) complexes to mice for three consecutive days and compared impacts on major leukocyte populations in secondary lymphoid organs versus in the gut and gut-associated lymphoid tissues. While we confirmed established patterns of IL-2-induced CD8 T cell and NK cell proliferation and expansion in the spleen and peripheral lymph nodes, these subsets were not impacted in the gut. Unexpectedly, we observed a sharp IL-2-dependent decline in Peyer’s Patch cellularity. This correlated with a specific reduction in B cells, while B cells in secondary lymphoid organs were not changed. IL-2 signals promoted selective and acute apoptosis in Peyer’s Patch B cells and phenotypic analysis revealed that IgD+ B cells were most impacted. Our results highlight the importance of fully elucidating systemic impacts of IL-2 complex administration, a widely used approach to modulate T cell and NK cell activity in diverse experimental settings, to properly interpret outcomes. This work may also aid in defining novel ways in which IL-2 contributes to the etiology, severity, and differential presentation of gastrointestinal conditions such as Crohn’s Disease.
Standardized Approach for Monitoring Plasma Cell Disorders
Authors: Johansson Ulrika , Li Na , Golts Natalie , Saksena Suraj 
Published: January 2023

Abstract
Multi-color flow cytometry using consensus surface markers (CD138, CD38, CD19, CD45, CD27, CD56, CD81, and CD117), intracellular kappa and lambda, as well as other auxiliary markers is routinely used for the identification and characterization of plasma cells. Here, we describe the development of a modular 10-color plasma cell panel for immunophenotyping of the plasma cell compartment. The panel was developed as an 8-color dried backbone, manufactured in a single test-per-tube, ready to use format. The dried backbone can be supplemented with liquid (drop in) reagents, which provides the flexibility of using intracellular kappa and lambda to examine clonality or using CD38-multi-epitope and/or anti-p63 (VS38c) reagent(s). The single test-per-tube format is ideally suited for MRD applications, where sample size is limited and acquisition of millions of events is needed for data analysis. In addition, the dried reagent tubes have a long shelf life (12 months at 20–25°C vs. 24 hours for liquid cocktails with multiple BV dyes in BSB buffer at 2–8°C). Furthermore, utilization of multiple high-performing polymer dyes in the panel affords high resolution of the relevant cell populations.
The dried plasma cell tube(s) were tested on bone marrow samples from healthy and diseased subjects in conjunction with intracellular staining for kappa and lambda. Our results demonstrate that the 10-color plasma cell panel can be used for identification of the plasma cell compartment (based CD38 and CD138 expression), assessment of aberrant marker expression (CD19, CD27, CD81, CD117, CD45, and CD56) for the identification of normal and abnormal plasma cells, and examining clonality in multiple patient samples (Kappa and Lambda).
Photobiomodulation (PBM) regulates nitric oxide (NO) production by peripheral blood mononuclear cells (PBMC) isolated from Multiple Sclerosis (MS) patients
Authors: Tolentino Miguel , Cho Chi C , Lyons Jeri-Anne 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an autoimmune disease characterized by myelin destruction, neurodegeneration and mitochondrial dysfunction. High production of nitric oxide (NO) by iNOS contributes to the generation of reactive nitrogen species, leading to increased nitrosative stress and mitochondrial dysfunction linked to MS progression. Current therapies focus on the suppression of the immune system in early stages, but they are not effective in chronic stages where nitrosative stress takes control over pathogenic mechanisms. Thus, new therapies that reduce nitrosative stress and offer neuroprotection at any stage are needed. Photobiomodulation (PBM) has demonstrated efficacy to improve neurodegenerative disorders. Our lab showed that PBM at 670nm reduced clinical severity in the C57BL/6 mouse experimental autoimmune encephalomyelitis model of MS by down-regulation of nitrosative stress, apoptosis, and pro-inflammatory cytokines, and up-regulation of anti-inflammatory cytokines. Likewise, PBM modulated cytokine expression by peripheral blood mononuclear cells (PBMC) and CD4+ T cells isolated from MS patients. Here we show that PBM reduces NO production by PBMC from MS patients. PBMC were isolated by gradient centrifugation, activated with Phytohaemagglutinin (PHA) and received PBM treatment. As surrogate for NO, nitrite production in supernatants was measured by fluorometry. The most effective PBM doses at decreasing nitrite were 830nm (10J/cm2) and pulsed 640nm (3J/cm2). The expression of Interferon-γ correlated with nitrite expression at these same doses. PBM effect was dependent on the disease modifying drug that subjects were taking. The results show the potential of PBM on NO regulation in MS patients.
Characterization of Immune Phenotypes in Peripheral Blood of Adult Renal Transplant Recipients using Mass Cytometry (CyTOF)
Authors: Kowli Sangeeta , Martinez Olivia M , Lebrec Herve , Minocherhomji Sheroy , Maecker Holden T. 
Published: January 2023

Abstract

Background
Chronic immunosuppressive therapy in transplant patients can be associated with opportunistic infections and risks of cancer development. There is an important need to better measure the immune status of patients in a transplant setting to improve patient monitoring and guide development of appropriate immunosuppressive agents and regimen.

Results
In this study, CyTOF was performed to comprehensively characterize the immune differences between 10 renal transplant recipients and 11 age-matched healthy donors, longitudinally over 6 months. Overall, the immune cells of transplant patients showed signs of increased CD8+ T cell activation/differentiation. Higher levels of CD57 on CD8+ T cells were observed. CD8+ T cells and their memory subsets also showed increased production of MIP-1β and IFNγ. Further, CD107a and Granzyme B expression were also increased in CD8+ T cells and CD56hi NK cells, while Tregs had decreased IL-10 production. These changes are consistent with a more stimulated immune system, despite the administration of immune suppressants to the patients. However, there were also some signs of immune suppression in the transplant patients consistent with the treatment regimen. γδT cells showed decreased TNFα and IFNγ expression, while MIP-1β expression was reduced in NKT, CD56hi NK and γδT cells. CD8+ T memory cell subsets also showed decreased expression of IL-2, IL-17 and GM-CSF.

Conclusions
This mosaic pattern of functional changes highlights the importance of comprehensive immune monitoring of these patients. Such information will aid in optimizing individual treatment strategies and development of improved immunosuppressants that specifically target overly activated populations.
Immunoproteomic identification of in vivo response toward residual antigenic proteins in xenogeneic heart valve biomaterials
Authors: Shortreed Nicholas A , Panicker Anjali J , Dalgliesh Ailsa J , Gates Katherine V , Griffiths Leigh G 
Published: January 2023

Abstract
Valve replacement is an important therapeutic approach for management of valvular heart disease, accounting for over 200,000 surgeries annually worldwide. Deficiencies of current heart valve prostheses led the NHLBI cardiac surgery working group to highlight the need for improved heart valve leaflet biomaterials. Recently, our group has engineered a novel method to eliminate antigens from bovine pericardium (BP) extracellular matrix (ECM) scaffolds, while retaining biomaterial structure-function properties and regenerative capacity. However, despite reducing BP-ECM scaffold antigen burden by &gt;92%, graft-specific humoral IgG response persisted at ~30% of the level associated with native BP in a rabbit model. In this study, we sought to determine the identities of residual antigens present in antigen removed BP-ECM scaffolds. New Zealand White Rabbits were subpannicularly implanted with either native, glutaraldehyde fixed, or antigen-removed BP-ECM scaffolds (n = 6 per group). Rabbit serum, collected pre- and at 56 d post-implantation, was used to generate IgG affinity chromatography columns. Native BP protein extracts were loaded onto columns, and captured antigens were identified by LC-MS/MS. A total of 259 antigens were identified in native BP rabbits. The glutaraldehyde fixed and antigen-removed rabbits responded to ~4% and ~12% of these same antigens, respectively. A small subset of antigens were uniquely identified in each of the treatment groups, not identified in native BP. These results inform ongoing efforts to eliminate antigens from xenogeneic tissues in development of ECM scaffolds, and may provide insight into the relative immunogenicity of individual xenoantigens in the context of tissue engineering.
TREX1-induced chromosome fragmentation at the interface of innate immunity and genomic instability
Authors: Mohr Lisa , Chu Kevan , Maciejowski John 
Published: January 2023

Abstract
The cGAS-STING pathway, a crucial component of innate immunity, recognizes viral DNA in the cytosol. cGAS binds dsDNA and catalyzes the production of the second messenger cGAMP, resulting in the expression of Type I interferons and other pro-inflammatory genes. To prevent autoimmunity, activation of cGAS by cytosolic self-DNA is normally counteracted by the cytosolic nuclease TREX1. In many chromosomally instable cancers, cytosolic self-DNA accumulates as DNA bridges or micronuclei (MN), whose nuclear envelopes are prone to rupture, exposing contained DNA to the cytosol. cGAS activation at ruptured MN is implicated in anti-tumor immunity. However, mechanisms regulating cGAS activation at ruptured MN are yet unknown. Here, we show that TREX1 partially degrades chromosomal DNA in the cytosol to suppress cGAS activation. We observed localization of cGAS and TREX1 to ruptured MN and DNA bridges, strongly suggesting an unknown role for TREX1 in regulating cGAS activation in cancer. After induction of MN or DNA bridge formation, TREX1 KO causes dramatic increases in cGAMP production and cGAS-STING pathway signaling. Furthermore, ruptured MN exhibit TREX1-dependent DNA damage and ssDNA foci. TREX1 is bound to the ER, which is known to invade ruptured MN. We found that an ER binding mutant of TREX1 does not localize to MN anymore and was not able to rescue the TREX1 KO phenotype, suggesting a role for the ER to recruit TREX1 to MN.
In summary, we propose that TREX1 enters MN as a consequence of nuclear envelope rupture to partially degrade DNA, thereby suppressing cGAS activation. By eliminating the immunogenic potential of these DNA species, TREX1 may dampen anti-tumor immunity and thereby help cancer cells escape the immune system.
Enhanced anti-tumor response upon deficiency of Msn kinase MINK1 in mice
Authors: Zhu Kaixiang , Lu Linrong 
Published: January 2023

Abstract
Misshapen (Msn)/NIK-related kinase 1 (MINK1) is a serine/threonine kinase belongs to the germinal center kinase (GCK) family. Previous studies indicated the correlation of MINK1 with tumor cell senescence and Th17 differentiation. Its dual role in modulating both tumor growth and T cell function indicated MINK1 a possible intervention target in tumor immunotherapy. We have utilized the MINK1 deficient mice to explore the possible role of MINK1 in the anti-tumor immune response. By using the subcutaneous tumor-xenograft model, we have found that MINK1 deficiency in mice resulted in slower tumor growth. We have also accumulated evidence in adoptive transfer and CD8+ T cells depletion experiments to show that the genetic ablation of MINK1 suppressed tumor growth primarily via CD8+ T cells without affecting other immune cells. Along with which, MINK1 deficient CD8+ T cells are superior in both tumor infiltration and the expression of effector markers such as TNF-α, IFN-γ, and Granzyme-B when compared to WT CD8+ T cells. Lower expression of exhaustion markers such as PD-1, TIM-3, and TIGIT were also observed in MINK1 KO CD8+ T cells. We are currently exploring the possible mechanisms of how MINK1 regulated CD8+ T cell function, and examine the possibility to target MINK1 in experimental tumor immunotherapy.