bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD91-Dependent Release of IL-1β by GP96 Involves the Activation of the Inflammasome Complex
Authors: Wang Yifei , Binder Robert J 
Published: January 2023

Abstract
The immunogenic heat shock protein, gp96, binds to CD91 to elite an immune response, characterized by cross-presentation of chaperoned peptides and release of cytokines, including IL-1β. IL-1β is pro-inflammatory cytokine, which is secreted by immune cells upon sensing pathogen associated molecular pattern (PAMP) or damage associated molecular pattern (DAMP). IL-1β is synthesized as a precursor protein, and requires cleavage by Caspase-1 to the matured IL-1β which is released from the cells. Caspase-1 activation requires the inflammasome protein complex. For Inflammasome activation, signal 1 induces pro-inflammatory gene transcription, and signal 2 activates Inflammasome. We have previously shown that gp96 induces NFκB activation and IL-1β secretion via CD91.1 Here we examined the utilization of the inflammasome for IL-1β in this setting. We report that gp96 is able to work as signal 1 to prime production of pro-IL-1β in APCs and in a dose-dependent manner. The kinetics of priming pro-IL-1β differ between gp96 and other PAMPs such as LPS. We will explore the dependence of IL-1β release on the inflammasome.
Neonatal CD8+ T cells provide innate immune protection against unrelated pathogens
Authors: Watson Neva B , Peng Seth , Nzingha Kito , Wang Jocelyn , Grenier Jennifer K , Smith Norah L , Patel Ravi K , Grimson Andrew W , Rudd Brian D 
Published: January 2023

Abstract
We previously found that neonatal and adult CD8+ T cells adopt different fates after infection with specific pathogens because they are derived from a distinct progenitor cell that is distinguished by expression of Lin28b. In this report, we investigated if CD8+ T cells respond differently to unrelated pathogens in early life. To start, we compared gene expression profiles of neonatal and adult CD8+ T cells after stimulation with innate cytokines (IL-12 + IL-18). Interestingly, we found that neonatal CD8+ T cells are more responsive and express a distinct cytokine profile compared to their adult counterparts. To examine the biological significance of these age-related differences, we next adoptively transferred monoclonal populations of neonatal or adult CD8+ T cells into T cell-deficient recipients and compared their ability to mediate immune protection against an irrelevant strain of Listeria monocytogenes. Surprisingly, we found that neonatal donor cell displayed increased proliferation and effector functions, and the pathogen burden was significantly reduced when compared to recipients containing adult donor cells. We also found that Lin28b may be responsible for the enhanced responsiveness to innate cytokines in early life, as ectopic expression of Lin28b in adult cells enabled them to initiate a program of bystander activation that was analogous to neonates. Collectively, these findings indicate that neonatal CD8+ T cells possess innate-like functions during infections at the cost of generating memory, and these unique behaviors may be driven by high levels of Lin28b. We currently are employing ATAC-seq to determine whether Lin28b changes the program of bystander activation by altering the chromatin landscape.
TGF-β Controls the Formation of Kidney-Resident T Cells via Promoting Effector T Cell Extravasation
Authors: Mishra Shruti , Ma Chaoyu , Demel Erika L , Liu Yong , Zhang Nu 
Published: January 2023

Abstract
Tissue-resident memory T (TRM) cells, a population of noncirculating memory T cells, are one of the essential components of immunological memory in both mouse and human. Although CD69+CD103+ TRM cells represent a major TRM cell population in barrier tissues including the mucosal surface and the skin, CD69+CD103− TRM cells dominate most nonbarrier tissues, such as the kidney. TGF-β is required for the differentiation of CD69+CD103+ TRM cells in barrier tissues. However, the developmental control of CD69+CD103− TRM cells in nonbarrier tissues remains largely unknown and the involvement of TGF-β signaling is less clear. In this study we demonstrated that TGF-β promoted the formation of kidney-resident T cells via enhancing the tissue entry of effector T cells. Mechanistically, TGF-β enhanced E- and P-selectin and inflammatory chemokine-mediated extravasation of effector T cells. Thus TGF-β controls the first developmental checkpoint of TRM cell differentiation in nonbarrier tissues.
Diet-induced Obesity Impairs Murine B cell Development and Antibody Production Accompanied by a Defect in Production of SPMs
Authors: Crouch Miranda J , Guesdon William , Armstrong Michael , Reisdorph Nichole , Shaikh Saame Raza 
Published: January 2023

Abstract
Diet-induced obesity is associated with increased risk for infections and poor responses to vaccinations, which may be due to compromised B cell function. There is limited information about the effects of obesity on B cell function outside of the context of adipose tissue inflammation. Therefore, we studied B cell development and ex vivo B cell antibody production in a murine model of diet-induced obesity. Analysis of the bone marrow revealed that obese mice displayed a two-fold reduction in the number of CD19+ cells, resulting in decreased frequencies of various B cell subsets. Early lymphoid commitment markers such as IL7Rα, IL7R, and STAT5 were significantly decreased at the transcript level. In addition, obese mice had reduced mRNA expression of the B cell lymphopoiesis markers, PAX5 and Oct2, compared to controls. Functionally, splenic B cells from obese mice were hyperstimulated with elevated ex vivo IgM and IgG secretion in the absence of stimulation. When B cells from the obese mice were challenged with anti-TLR4 or anti-BCR/TLR9 in culture, IgM and IgG levels were diminished. Mechanistically, the levels of D-series specialized pro-resolving lipid mediators (SPMs), which are potent regulators of inflammation and adaptive immunity, were lowered in obese mice compared to controls. Furthermore, select enzymes involved in generation of SPMs were also decreased in obese mice. Overall, these findings demonstrate that obesity hinders B cell development and drives a dysregulated antibody response, which could contribute to impaired responses to infections and vaccinations.
The Toll-like receptor ligand CpG-A induces type 1 interferons in B cells contrasting the proinflammatory inducing activity of CpG-B
Authors: Akkaya Munir , Akkaya Billur , Sheehan Patrick , Miozzo Pietro , Rawat Mukul , Pena Mirna , Kim Ann S , Kamenyeva Olena , Kabat Juraj , Qi Chen-Feng , Bolland Silvia , Chaturvedi Akanksha , Pierce Susan K 
Published: January 2023

Abstract
B cells express the innate immune system receptor, Toll like receptor 9 (TLR9), a potent regulator of B cell function, that signals in response to unmethylated CpG sequences in microbial DNA. A dichotomy in the function of the two major classes of CpG-containing oligonucleotides, namely CpG-A and CpG-B, appears to exist with CpG-A restricted to inducing type 1 interferon (type 1 IFN) in innate immune cells and CpG-B to activating B cells to proliferate and produce antibody and inflammatory cytokines. Although CpGs are candidates for adjuvants to boost the innate and adaptive immunity, our understanding of the effect of CpG-A and CpG-B on B cell responses is incomplete. We provide evidence that CpG-A activated B cells in vitro in a TLR9 dependent fashion resulting in the production of IL-6, proliferation, and alterations in the expression of a variety of B cell surface markers, albeit in a less robust fashion as compared to CpG-B. Neither CpGB nor CpG-A alone induced B cells to express type 1 IFN. However, when incorporated into the cationic lipid, DOTAP, CpG-A, but not CpG-B induced a robust type 1 IFN response in B cells both in vitro and in vivo. Conversely, CpG-B-DOTAP but not CpGA-DOTAP promoted B cell germinal center responses in vivo and the production of high affinity antibodies. We also provide evidence that difference in the function of CpGs versus DOTAP-associated-CpGs may be related to their intracellular trafficking in B cells since DOTAP enhances the trafficking of CpG-B but not CpG-A to late endosomal compartments. These findings have important implications for the use of CpG-A and CpG-B to augment type 1 IFN production versus B cell responses in vivo.
Exosomes secreted from virus- infected microglia can activate an inflammatory response in the central nervous system
Authors: Olson Julie K , Luong Nhungoc 
Published: January 2023

Abstract
Exosomes are membrane-bound nanovesicles released from cells that can be taken up by other cells, thus mediating intercellular communication. Exosomes contain miRNA, mRNA, and proteins that are specific to the cells from which they were released. Theiler’s murine encephalomyelitis virus (TMEV) induced demyelinating disease is a mouse model for multiple sclerosis. TMEV infection of susceptible mice leads to a persistent viral infection of microglia in the central nervous system (CNS) which contributes to the development of a chronic progressive demyelinating disease associated with an inflammatory immune response. We have previously shown that TMEV infection of microglia activates an innate immune response with expression of type I interferons, cytokines, chemokines, and effector molecules associated with development and progression of demyelinating disease. Next, we wanted to determine whether microglia infected with TMEV secrete exosomes and whether these exosomes could influence uninfected CNS resident cells, such as microglia, astrocytes, and neurons. Our studies show that exosomes secreted from virus-infected microglia contain viral RNA that can activate an innate immune response in uninfected bystander cells which leads to increased expression of pro-inflammatory cytokines. Most interestingly, exosomes secreted from microglia in the brain of TMEV- infected mice could be transferred to uninfected mice and activate an inflammatory immune response in the brain of uninfected mice. These results suggest that exosomes secreted by virus-infected microglia can activate the immune response in bystander uninfected cells which may contribute to the inflammatory response associated with demyelinating disease.
The role of coinhibitory pathways in allergen-specific CD4+ T cell responses
Authors: Rosskopf Sandra , Jahn-Schmid Beatrice , Bohle Barbara , Battin Claire , Schmetterer Klaus , Pickl Winfried F , Steinberger Peter 
Published: January 2023

Abstract
More than 25% of the population suffers from IgE-mediated immune reactions and its related symptoms. T lymphocytes have a crucial role in initiating and promoting allergies and their responses are tightly regulated by numerous activating and inhibitory signals provided by APCs. Currently, there is limited knowledge regarding the role of inhibitory pathways in allergen-specific CD4+ T cell responses. To address this issue, patients allergic against house dust mites, birch pollen, mugwort pollen and grass pollen were recruited. PBMCs were isolated and then stimulated with allergenic extracts to evaluate coinhibitory pathways using blocking antibodies (e.g. nivolumab, ipilimumab) and to assess the expression of coinhibitory receptors on allergen-specific T cells. Allergenic extracts were analyzed for presence of TLR triggering molecules using NF-κB-eGFP reporter cells expressing TLR1/2, TLR2/6, TLR4 or TLR5. Furthermore, allergen-specific T cell reporters, T cell lines or T cell clones from allergic patients were cocultivated with engineered APCs that present allergenic peptides on MHC class II molecules and express different coinhibitory molecules. Blockade of the PD-1/PD-L1 axis strongly enhanced proliferation of allergen-specific CD4+ T cells in response to allergenic extracts. No correlation was found between IgE concentration and T cell proliferation. Preliminary results show a distinct expression pattern of coinhibitory molecules on allergen-specific T cells. In a next step Th1/Th2 cytokine patterns will be analyzed. Research was performed in accordance with the Declaration of Helsinki and approval of the local ethics committee was obtained (EK1538/2014).
House dust mite allergen induces inflammasome activation and NFkB signal transduction in monocytes
Authors: Yu Sheng-Jie , Yen Hung-Rong 
Published: January 2023

Abstract
Over the past few decades, allergic diseases have become increasingly prevalent worldwide. House dust mite (HDM) is the most important domestic source for allergic diseases such as allergic rhinitis, asthma and atopic dermatitis. Dermatophagoides pteronyssinus (Der p) is the major environmental allergen in southeast Asia because of the humid and warm environment is suitable for its growth. Der p has been identified which could increase proinflammatory cytokine expression through activating innate immune response. Inflammasomes activation was reported which could help host defense pathogens and allergen in the recent years. However, the correlation between HDM induced allergic inflammation and inflammasomes activation remains unclear. The aim of this study was to investigate the effect of HDM allergen on inflammasomes activation and downstream signal transduction. We found that THP-1 cells, stimulated with HDM crude extract, activated NALP1, NLRC4, NLRP3 and AIM2 inflammasomes in a time-dependent manner. The expression of adaptor protein ASC and its caspase effector, Caspase-1, as well as proinflammatory cytokines, IL-1β, IL-6, IL-8 and TNF-α were significantly upregulated after HDM stimulation. Mechanistic study revealed that HDM stimulation increased the expression of phosphorylated NFkB, JNK and AKT. Our study suggests that HDM allergen could induce inflammatory response through NFkB signal transduction pathway and inflammasomes activation. Thus, we could use these data for developing specific target immunotherapy in the future.
A cross species CD200R1 immune checkpoint agonist with potent anti-inflammatory properties
Authors: Gariepy Jean , Khatri Ismat , Gorczynski Reginald , Young Ashley , Woo Lindsay , Chow Chung-Wai , Cydzik Marzena , Prodeus Aaron 
Published: January 2023

Abstract
Functional aptamers displaying agonistic or antagonistic properties are showing great promise in terms of modulating immune responses. Our group has recently developed pegylated DNA aptamers that can either block inflammatory responses (CD200R1agonist) or restore tumor-directed immune responses (PD-1 antagonist) in vivo. Here, we report the derivation and design of a cross species mouse/human CD200R1 DNA aptamer agonist that blocks inflammatory responses in mouse models of skin graft rejection and asthma. This DNA aptamer was discovered by performing NGS and comparing the resulting aptamer motifs derived from independently screening mouse and human CD200R1 as targets. Importantly, this m/hCD200R1 agonistic aptamer does not suppress cytotoxic T-lymphocyte (CTL) induction in 5 day allo-mixed lymphocyte cultures (MLCs) derived from CD200R1 knockout mice, indicating that its mode of action is directly linked to CD200R1 activation. This study suggests that one can derive agonistic DNA aptamers that can be verified as immunomodulators in mouse models with outcomes translatable to the treatment of human conditions.
Class switch recombination in the absence of the IgH 3′ regulatory region
Authors: Yu Kefei , Kim Ahrom , Han Li 
Published: January 2023

Abstract
The ~28 kb 3′-regulatory region (3′RR) which is located at the most distal 3′ region of the immunoglobulin (Ig) heavy (H) chain locus has multiple regulatory functions that control IgH expression, class switch recombination (CSR) and somatic hypermutation. Here, we report that deletion of the entire 3′RR in a mouse B cell line that is capable of robust cytokine-dependent CSR to IgA results in reduced but not abolished CSR. These data suggest that the 3′RR is not absolutely required for CSR, and thus is not essential for targeting activation-induced cytidine deaminase to switch regions as has been suggested. Moreover, replacing the 3′RR with a DNA fragment including only its four DNase I hypersensitive sites (lacking the large spacer regions), restores CSR to a level equivalent to or even higher than wild type cells, suggesting that the four HS sites contain most of the CSR-promoting functions of the 3′RR. Stimulated cells express abundant germline transcripts, with the presence or absence of the 3′RR, providing evidence that the 3′RR has a separate role in promoting CSR, that is unique from enhancing S region transcription.
Visualization of binding of RAG2 to H3K4me3 regulated by its noncore domain.
Authors: Rodgers William A , Byrum Jennifer , Rodgers Karla K 
Published: January 2023

Abstract
Association of RAG2 with the epigenetic marker H3K4me3 through a PHD domain in its noncore region is important for activating the RAG1 component of the RAG recombinase complex for V(D)J recombination. However, immunofluorescence staining of RAG2 expressed in thymocytes suggests RAG2 is localized principally to the nuclear periphery, away from sites of H3K4 modification. To further test for interactions between RAG2 and separate epigenetic markers, we used confocal microscopy to measure full length RAG2 labeled with GFP at its N-terminus (GFP-FL) and expressed in RAG2−/−pro-B cells. Cross-correlation analysis of cells immunostained with antibodies to H3K4me3 and H3K9me3 showed that RAG2 colocalization with H3K4me3 was significantly greater than with H3K9me3. Furthermore, blocking histone demethylation by treating cells with 2,4-pyridinedicarboxylic acid increased colocalization of GFP-FL with H3K4me3, but not its colocalization with H3K9me3. Interestingly, a GFP-labeled T490A mutant of full length RAG2 (GFP-T490A) showed significantly greater colocalization with H3K4me3 than the labeled wild-type RAG2. GFP-labeled RAG2 core alone (GFP-core) exhibited the poorest colocalization with H3K4me3 of the constructs that we measured. Finally, nanoscale resolution using structured illumination microscopy showed that RAG2 was unevenly distributed in the nucleus, containing dispersed RAG2-rich and RAG2-poor regions. Furthermore, RAG2 interactions with H3K4me3 were restricted to interfaces between H3K4me3-enriched puncta and the RAG2. Altogether, these data provide visualization of RAG2 binding to the epigenetic marker H3K4me3, and that Thr490 of RAG2 regulates this interaction.
The microRNA processing subunit DGCR8 is required for immune response and germinal center formation
Authors: Daum Patrick , Meinzinger Julia , Pracht Katharina , Côrte-Real Joana , Wittmann Jürgen , Jäck Hans-Martin 
Published: January 2023

Abstract
MicroRNAs (miRNA) are a family of single stranded non-coding RNAs that regulate gene expression at the post-transcriptional level. The microprocessor complex subunit DGCR8, which is pivotal for the canonical processing of precursor transcripts into mature miRNAs, has been shown to be indispensable for the pro-B to pre-B cell transition in early B cell development (Brandl et al., 2016). To understand the role of DGCR8 and miRNA processing during the antigen-dependent activation of mature B cells, we deleted DGCR8 during the development of immature IgM-positive B cells into mature splenic B cells by crossing a transgenic CD23-Cre mouse with a mouse homozygous for floxed DGCR8 alleles.
Analysis of LPS-activated DGCR8-deficient splenic B cells from these mice revealed a reduced abundance of mature miRNAs, impaired in vitro survival, and reduced class switch to IgG1. Furthermore, frequencies of follicular B cells in the spleen, as well as B2 B cells in the peritoneal cavity were significantly altered. Finally, mice with a B cell-specific DGCR8-deficiency failed to produce antigen-specific serum IgG antibodies and adequate numbers of GL7-positive germinal center B cells upon immunization with a thymus-dependent antigen.
In summary DGCR8, and thus, most likely miRNAs are required to mount effective and specific T cell-mediated immune responses.
Gene conversion contributes significantly to IgHV somatic hypermutation in mice and humans
Authors: Dale Gordon A , Bohannon Caitlin D , Wilkins Daniel J , Dilernia Dario , Hunter Eric , Bedford Trevor , Antia Rustom , Sanz Ignacio , Jacob Joshy 
Published: January 2023

Abstract
During antigen-driven affinity maturation germinal center B lymphocytes undergo multiple rounds of somatic mutagenesis in the rearranged IgHV gene segment. A direct consequence of this process is the generation of multiple antibody mutants that are, in general, specific for antigen. Survival of these mutants is contingent on preserving or improving antigen-specificity in the complementarity-determining regions (CDRs) as well as retaining a functional framework region (FWR). Broadly, somatic mutations in germinal center B cells can be classified as templated or untemplated. In animals such as rabbits and chickens, somatic mutations are mostly templated. Conversely, in mice and humans it is widely accepted that mutations are untemplated. Here, we argue in favor of the alternative, that a predominant number of somatic mutations in IgHV gene segments in mice and humans can be traced to germline IgHV genes and occur through gene conversion. Using the NP hapten model, we demonstrate that mutations in germinal center B cells and plasma cells are traceable to other germline IgHV genes. Subsequently we also demonstrate using the novel V-LAIR1-DJ antibodies described by Tan et al (Nature 2016) that mutations in human antibody IgHV rearrangements, including those in non-immunoglobulin sequences (LAIR1), are traceable to the human IgHV germline repertoire. Together, this suggests that the plethora of diversity generated in antigen specific B cells is primarily directed by the germline repertoire of IgHV genes.
CD8+ tissue resident memory T cells are associated with good prognosis in breast cancer patients
Authors: Egelston Colt , Srinivasan Gayathri , Avalos Christian , Huang Yinghui , Rosario Anthony , Wang Roger , Jimenez Grecia , Simons Diana L , Yost Susan , Yuan Yuan , Lee Peter P 
Published: January 2023

Abstract
CD8+ Tumor Infiltrating Lymphocytes (TILs) have been shown to correlate with patient prognosis in breast cancer. Recently, resident memory T cells (TRM) have been established as a potent functional subset of memory T cells that exist in peripheral tissue without recirculation. We assayed CD8+ T cells from fresh breast tumors to phenotype memory T cells and found them to be almost exclusively made up of effector memory T cells. Further profiling by examining expression of CD103 and CD69 showed that on average 40% of CD8+ TILs are composed of TRM. Functional analysis of these T cells showed robust production of IFNγ and TNFα by these TRMs suggesting that despite existence in the tumor microenvironment TRMs maintain functional potency. To examine if CD8+ TRM have prognostic value in breast cancer patients, we examined a set of primary tumors from ‘good’ and ‘bad’ outcome patients, which we defined as having a relapse in more than 5 years after diagnosis or in less than 3 years after diagnosis respectively. In good outcome patients TRMs represented 60% of CD8+ T cells found in cancer islands, while only 20% of such in bad outcome patients. Similarly, CD8+ TRMs were observed in higher numbers in both the cancer islands and stroma of good outcome patient tumors in comparison to bad outcome patient tumors. This observed significance was greater than that found for CD8+ T cells in general, highlighting the importance of TRM in affecting patient outcome. Finally we showed that TRMs also exist in non-cancerous breast tissue and are found within the mammary ducts associated with epithelial cells perhaps in surveillance of potential pathogens. We suggest that TRMs are a potential vaccine target for preventing recurrence and metastasis in cancer patients.