bluuebunny/crossref_metadata_2025_split huggingface dataset:
Innate immune responses in fungal asthma enhance antiviral immunity during influenza virus infections
Authors: Samarasinghe Amali , LeMessurier Kim , Doorley Laura , Palipane Maneesha 
Published: January 2023

Abstract
Fungal allergic asthma is a subtype of asthma with high incidence. Although asthmatics are considered an ‘at risk’ group for severe influenza, these patients suffered less severe morbidity and mortality compared to non-asthmatics during the 2009 influenza pandemic. We developed a mouse model of fungal asthma and influenza comorbidities to recapitulate these epidemiologic findings and provide a means to explore mechanisms by which this protection may occur. We hypothesized that eosinophils guide antiviral responses against influenza virus infections in the allergic host. We characterized the immunophenotype of eosinophils in various niches including bone marrow, lymphoid organs, and lungs, in asthma and influenza comorbid conditions in comparison with individual disease conditions. Eosinophils from comorbid mice had high expression of MHCI, PIR-A/B, and CD62L in the lungs and spleens compared to influenza alone. The adoptive transfer of eosinophils led to a reduction in viral burden and increase in airway CD8+ T cells in virus-infected recipients. In addition, eosinophils were able to promote CD8+ T cell activation in vitro and ex vivo, and antigen-loaded eosinophils interacted with CD8+ T cells in vitro. Comorbid mice had more plasma cells in lymphoid organs although mature B cell numbers were similar to influenza alone. Antibody production in these cells was more prominent at early time points in comorbidity but was similar between the groups at late time points. Since eosinophils have been previously shown to regulate B cells and IgA production in the gut, we are now exploring the hypothesis that eosinophil-mediated B cell responses contribute to the increase in neutralizing antibodies noted in mice with asthma and influenza.
Testing the hypothesis that malaria-induced immune remodeling of monocytes plays a role in immunity to malaria
Authors: GUHA RAJAN , Crompton Peter D. 
Published: January 2023

Abstract
In malaria-naïve individuals, Plasmodium falciparum infection results in high levels of parasite-infected red blood cells (iRBCs) that trigger the systemic inflammation and fever characteristic of malaria. But, individuals in endemic areas who are repeatedly infected with P. falciparum are often asymptomatic and have low levels of iRBCs, even young children who have yet to acquire fully protective antibodies. The molecular mechanisms underlying these clinical observations remain unclear. We earlier showed that PBMCs collected from uninfected Malian children before the malaria season responded to iRBCs by producing pro-inflammatory mediators such as IL-1β, IL-6. However, following febrile malaria there was a marked shift in the response to iRBCs with the same children’s PBMCs producing lower levels of those cytokines. Moreover, genome-wide expression analysis showed that genes involved in phagocytosis and intracellular killing were upregulated in PBMCs after malaria as compared to before. Together, these data suggest that malaria-induced remodeling of innate immune cells might play a role in immunity to malaria. In ongoing work we are dissecting the underlying molecular pathways involved in P. falciparum-induced changes in monocyte/macrophages. We are studying monocytes ex vivo from malaria-exposed children, as well as monocytes stimulated with iRBCs in vitro. Initial data support the hypothesis that malaria-induced immune-remodeling of monocytes dampens pathogenic inflammation while enhancing anti-parasite effector mechanisms, consistent with our observation that children in malaria endemic areas are often afebrile and tend to control parasite replication in the face of repeated P. falciparum exposures.
Microbiota-independent mechanisms shape Th17 homeostatic responses at the oral barrier.
Authors: Dutzan Nicolas , Abusleme Loreto , Bridgeman Hayley , Greenwell-Wild Teresa , Zangerle-Murray Tamsin , Fife Mark E , Bouladoux Nicolas , Linley Holly , Brenchley Laurie , Wemyss Kelly , Trinchieri Giorgio , Diaz Patricia I , Belkaid Yasmine , Konkel Joanne E , Moutsopoulos Niki M 
Published: January 2023

Abstract
At barrier sites, resident immune cell populations help to maintain tissue homeostasis and function. These cells receive and integrate key signals from the local environment including stromal/epithelial cells and the commensal microbiome. Studies of the skin and gastrointestinal tract have revealed the importance of these signals for the development of host immune response. However, which commensal or tissue-specific cues are important for the immune system at the oral barrier remains minimally explored. Th17 cells have been described as key mediators of immunity at the oral barrier but also essential for periodontitis, a highly prevalent inflammatory pathology that affects the gingiva. In this study we focused in the identification of the mechanisms controlling the induction and regulation of Th17 cells in the gingiva. Our data show that IL-17-producing CD4+ T cells increase with age and their accumulation at the oral barrier occurs independently of commensal colonization. Moreover, we demonstrate that IL-6 elicited by physiological mechanical damage during mastication shapes the function of T cells at the oral mucosa, promoting Th17 differentiation. Finally, we observe that long-term mechanical damage through mastication induces IL-17 mediated bone loss at the gingival barrier. Our data highlight the notion that a variety of signals may be essential to shape the immune responses at different barrier sites, and particularly at the oral cavity, unique mechanisms modulates homeostatic and also pathogenic Th17 responses.
A donor-specific primary monocyte-derived dendritic cell maturation model for high throughput screening of immunostimulatory compounds
Authors: Kelly Erin , Lawrenz Anthony , Wong Alan , Harberts Erin M , Ernst Robert K , Shirazian Afshin , Chen Qin 
Published: January 2023

Abstract
Dendritic cell (DC)-based immunotherapy is a promising treatment alternative for drug-resistant malignancies. Identifying candidate immunostimulatory compounds that can reliably facilitate DC maturation in the context of antigen-specific therapies is a significant hurdle in the development of such immunotherapies. In this study, we present a high throughput in vitro primary cell assay to evaluate dose-dependent DC maturation in a panel of healthy donor primary cells. CD14+ monocytes from four healthy donors were isolated from leukapheresis (Leuko Pak)-derived peripheral blood mononuclear cells (PBMC), differentiated by culture with GM-CSF and IL-4 cytokines, and then stimulated with lipopolysaccharide (LPS), CpG oligonucleotides, or a panel of novel Toll-like receptor 4 (TLR4) ligands. To quantify the resultant maturation, flow cytometry was used to measure the expression of cell surface biomarkers MHC-II, CD40, CD80, CD83, and CD86. While monocytes from all donors responded to the ligands tested, the intensity of activation marker expression varied between donors. LPS-induction of CD83 expression ranged from 27 to 82% of DCs amongst the 4 donors tested, highlighting the importance of multi-donor screening in interpreting quantitative results. In conclusion, a high throughput in vitro primary monocyte-derived DC differentiation assay reveals donor to donor variations that may go unobserved in screening experiments completed in cell lines or single donor primary cells sources. This assay can be used to assist researchers in lead generation, lead optimization, and mechanism of action studies of immunostimulatory compounds.
The role of Krueppel-like factor 2 in B cell activation and plasma cell homeostasis
Authors: Schuh Wolfgang , Schubert Tina , Knipfer Lisa , Wallrapp Antonia , Jäck Hans-Martin 
Published: January 2023

Abstract
Upon contact to antigen, B cells differentiate into memory B cells or antibody secreting short-lived or long-lived plasma cells (PC). The survival of long-lived PCs depends on interaction with survival niches in the spleen or in the bone marrow. However, the molecular mechanisms controlling PC generation, migration, homing and survival are poorly understood. We recently reported that Krueppel-like factor 2 (KLF2) is an important regulator of PC homeostasis. KLF2-deficient B cells express lower levels of homing factors, such as beta7-Integrin and L-Selectin. Numbers of antigen-specific PCs in the bone marrow of immunized KLF2-deficient mice are drastically reduced, indicating that KLF2 plays a crucial role in controlling PC homing and/or survival. Based on these findings we wanted to further characterize the functional impact of KLF2 on B cell activation and PC homeostasis. We observed that KLF2-deficient follicular B cells show a lower activation threshold and differentiate faster into plasma blasts in response to lower LPS concentrations in vitro compared to wildtype B cells. Furthermore, KLF2-deficient B cells proliferate faster as measured by CFSE. Target genes and signalling pathways modulated by KLF2 in B cells and PCs are currently analysed by gene expression profiling. We conclude that KLF2 acts as a quiescence factor in mature B cells by setting their activation threshold. In addition, KLF2 is crucial for PC homeostasis. KLF2-mediated signalling pathways and KLF2-regulated target genes which set the threshold for B cell activation, regulate plasma blast homing and mediate plasma cell survival will be identified by transcriptome analyses. This study was supported by DFG grants FOR832 and TRR130 to H.-M.J. and ELAN grant to W.S.
Role of Trim13 in toll-like receptor 2-mediated NF-κB activation
Authors: Baek Suk-Hwan , Huang Bin , Pei Han Zhong 
Published: January 2023

Abstract
NF-κB regulates the expression of a various genes involved in diverse cellular processes including inflammation and immunity. Ubiquitination is a versatile post-translational modification involved in NF-κB activation of toll-like receptor (TLR) signaling. Here, we demonstrated that Trim13, an E3 ubiquitin ligase, is up-regulated in macrophages upon stimulation with TLR2 ligand. Knock-down of Trim13 attenuated TLR2-mediated production of cytokines/chemokines and formation of foam cells, as well as activation of NF-κB. Trim13 interacts with TRAF6 and potentiates NF-κB activity via ubiquitination of TRAF6. Overexpression of inactive mutant (C10/13A) or RING deletion mutant of Trim13 did not potentiate ubiquitination of TRAF6 or activation of NF-κB. These results suggest that the effects of Trim13 are dependent on its E3 ligase activity. Trim13 used K29-linked polyubiquitin chains for TRAF6 ubiquitination to promote NF-κB activity and thus potentiated activation of TLR2-mediated immune responses. Our data identify Trim13 as a positive regulator of NF-κB activation and suggest that K29-linked polyubiquitination is a specific ubiquitin-linked pattern involved in the control of TLR2 signaling.
APOBEC gene family expression and hallmarks in chronic lymphocytic leukemia
Authors: Chu Charles C , Vergani Stefano , Yan Xiao-Jie , Dhayalan Arvind , Patten Piers E.M. , MacCarthy Thomas , Yuan Chaohui , Barrientos Jacqueline C. , Kolitz Jonathan E. , Allen Steven L. , Rai Kanti R. , Chiorazzi Nicholas 
Published: January 2023

Abstract
The hallmark activity of APOBEC (apolipoprotein B mRNA editing enzyme, catalytic polypeptide) family of cytidine deaminases, including activation-induced deaminase (AID) and APOBEC3 genes, has been detected in somatic mutation signatures by ultra-deep sequencing of the genomes of many cancers, including chronic lymphocytic leukemia (CLL). The acquisition of these mutations is hypothesized to lead to the progression towards aggressive disease in cancer. To examine this in CLL, we tested if increased APOBEC family member gene expression in CLL cells, as measured by microarray and quantitative real time PCR, correlated with worse patient outcome. Higher levels of AID, APOBEC3B, APOBEC3F and APOBEC3H in CLL cells correlated with worse patient outcome, whereas APOBEC3G did not. Interestingly, higher levels of a truncated splice variant of APOBEC3F tended to correlate with better patient outcome. The expression of truncated APOBEC3F may possibly interfere with APOBEC family member mutation activity. To test mutation activity, CLL cells were activated by transfer into NOD-scid IL2Rγnull mice, a xenograft model of human CLL, and hallmark mutation signatures in the expressed immunoglobulin variable region (IGHV) of CLL cells were analyzed by targeted ultra-deep sequencing. Induced IGHV mutation hallmarks consistent with AID were found. These data support the hypothesis that expression and mutation activity of APOBEC family members, such as AID, in CLL cells could lead to adverse patient consequences.
Antibody repertoire analysis in sera of breast cancer patients using a random peptide microarray differentiates cases from controls with high specificity and sensitivity
Authors: Chapoval Andrei I , Podlesknyh Stepan , Kolosova Evgenia , Anisimov Daniil , Ryzanov Mikhail , Petrova Valentina , Shcherbakov Dmitry , Johnston Stephen , Lazarev Aleksander 
Published: January 2023

Abstract
Breast cancer (BC) is the most common malignancy and the second most common cause of cancer-related mortality in women. Early detection of BC using different biomarkers is a key factor for successful BC therapy. Many studies have shown that serum antibodies (Ab) against tumor-associated antigens (TAAs) could be used as potential biomarkers in the detection of BC. However in some cases the most of informative TAAs for circulating Ab detection, at early stages of BC, might be unknown. Even if informative Ab targets are currently unknown it is possible to analyze Ab repertoire (immunosignature) using random peptide arrays. In the present study, we probe a custom 330,000 random 12-mer peptide microarray with sera from 42 BC cases and 41 controls to develop a diagnostic signature. We determined that differentiation of BC cases from controls with high sensitivity and specificity requires a panel of at least 100–150 peptides. Basic Local Alignment Search Tool (BLAST) searches suggest that the identified peptides have no similarity with known TAAs. However, despite the random nature of the amino acid sequence that were used in microchip design, common amino acid motif is present in the groups of peptides that react with sera Ab from patients with BC but not healthy control. Thus the evaluation of Ab repertoire (immunosignature) using microarray containing peptides with random amino acid sequences has a potential value in the early diagnosis of cancer.
Dietary Early Glycation Products Promote Prostate Tumorigenesis through Modulating Macrophages
Authors: Chen Yingjia , Guo Tai L 
Published: January 2023

Abstract
Glycation products are ubiquitous in food. Moreover, the well-controlled glycation, that results in the production of early glycation products (EGPs), has been proposed as a strategy to improve the physicochemical properties of the food proteins. However, the health effects of EGPs are rarely known. Since there is evidence showing that Western diet (glycation prone) is associated with higher mortality of prostate patients than Prudent diet, we investigated the role of EGPs in prostate tumorigenesis.
EGPs were produced from a whey protein isolate-glucose model system, with non-reacted samples (NR) and advanced glycation end-products (AGEs) as controls. They were used to gavage male C57BL6 mice, and the mice were then subcutaneously injected with TRAMP-C2 cells (a mouse prostate cancer (PCa) cell line) in the flank area. The tumor length, width, and the proliferation index were measured, and the splenic leukocytes were studied. In addition, the mouse TRAMP-C2 cells and two human PCa cell lines LNCaP and PC-3 were cultured with NR, EGPs and AGEs, and LNCaP and PC-3 were further cultured with the conditioned medium generated from human macrophages. The PCa cell proliferation, and the cytokine/chemokine production and iNOS expression by macrophages were determined. The results showed that EGPs promoted prostate tumor growth most in vivo, and PCa cell proliferation only when the presence of macrophages in vitro. In addition, EGPs activated the production of most cytokines/chemokines examined, with IL-10 and MIP-1β unregulated most, and decreased the iNOS expression in macrophage.
In conclusion, EGPs promoted PCa possibly by modulating macrophages to become prostate tumor-associated M2, which produced high levels of IL-10 and MIP-1β.
Enhanced inhibition of chemotaxis by adenosine in peripheral blood CD8+ T cells of head and neck cancer patients
Authors: Chimote Ameet A , Arnold Michael J , Balajthy Andras , Newton Hannah , Wise-Draper Trisha , Conforti Laura 
Published: January 2023

Abstract
Adenosine (Ado) accumulates in high concentrations in solid tumors where it contributes to the failure of the immune cells to eliminate cancer cells. We have previously shown in healthy T cells that Ado acts via A2A receptors and inhibits T cell motility by suppressing KCa3.1 channel function. Herein, we conducted experiments to elucidate the effect of Ado on the chemotactic abilities of peripheral blood T cells (PBTs) from head and neck squamous cell carcinoma (HNSCC) patients. CD8+ PBTs were isolated from HNSCC patients prior to any form of treatment and healthy donors (HD), and activated with CD3/CD28 antibodies. CD69 expression indicated that comparable activation was achieved in HNSCC and HD T cells. Experiments to measure chemotaxis towards a CXCL12 gradient were performed on T cells suspended in a collagen matrix, which resembles the fibrous network within solid tumors. Chemotaxis was evaluated by determining the center of mass along the Y-axis (Y-COM), which reflects the location of the cells at the end of the experiment and is indicative of their ability to move towards the chemokine gradient. PBTs of HNSCC patients and HDs underwent comparable chemotaxis in the CXCL12 environment. Next, we compared their chemotaxis when Ado was added to the chemokine gradient. Ado was responsible for a significantly greater reduction in the Y-COM in HNSCC (~82%, n=16) compared to HD (~26%, n=7; p=0.012). This effect was mimicked by the A2A receptor agonist CGS21680 and was reversed by the A2A receptor antagonist SCH5826. Our data suggest that Ado acts via A2A receptors and suppresses chemotaxis in HNSCC patients to a greater degree than their healthy equivalents, which may limit their ability to effectively penetrate the tumor.
Bacterial modulation of noncanonical inflammasome.
Authors: Vanaja Sivapriya Kailasan , Nair Meera Surendran , George Mariam , Rathinam Vijay 
Published: January 2023

Abstract
Sensing of microbial products by pattern recognition receptors to mount effective antimicrobial responses is an evolutionarily conserved immune strategy. Cytosolic LPS sensing by the noncanonical inflammasome via caspase-11 and the ensuing IL-1 and cell death responses are emerging as a key host defense strategy that play a crucial role in the clearance of infectious agents. A strong selection pressure from the host such as this drives pathogens to develop strategies to actively antagonize or evade inflammasome responses. However, it is not known if bacteria antagonize noncanonical inflammasome pathway. This study addresses this knowledge gap and examines the modulation of noncanonical inflammasome by bacteria utilizing Enterohemorrhagic E. coli (EHEC) as a model organism. Supporting the idea of inflammasome suppression by EHEC, our results indicate that compared to E. coli K12, an avirulent laboratory strain of E. coli, EHEC induces substantially weaker cell death and IL-1 secretion. Importantly, by screening a library of EHEC mutants lacking known and putative virulence factors, we found that a mutant that lacks an EHEC-specific genomic island triggered markedly higher cell death and IL-1 responses compared to wild-type EHEC revealing that this genomic island harbors a suppressor of noncanonical inflammasome. Altogether, this study attributes an inflammasome modulatory function to an EHEC genomic island, whose function was previously unknown. Identifying novel mechanisms by which pathogenic bacteria silence noncanonical inflammasome is crucial as it may aid in designing novel therapeutic approaches against Gram-negative infections.
Analysis of immunogenicity of different sizes of HER2/neu antigen
Authors: Jeon Insu , Seo Hyungseok , Park Myung Hwan , Shin Kwang-Soo , Lee Jeong-mi , Choi Jeongwon , Song Boyeong , Koh Choong-Hyun , Bae Eun-Ah , Kang Chang-Yuil 
Published: January 2023

Abstract
One of the most important factors of cancer immunotherapeutic vaccine is to induce tumor antigen specific T cell response. Prior to considering how to induce T cell response efficiently, researchers should select tumor antigen with careful consideration because some tumor antigen show oncogenic property that may lead to safety problems. To reduce these concerns, tumor antigens are used in artificially mutated or truncated form so that oncogenic function would be eliminated. On the other hand, as the size of antigen determines T cell receptor epitope repertoire, it is important to maintain the original size as much as possible. In this study, we tested immunogenicity of four different sizes of HER2/neu antigens in B cell and monocyte based vaccine models. In murine models, we observed antigen specific T cell response in the antigen size dependent manner and antigen specific antibody response in the antigen expression dependent manner. In addition, all of four different sizes of antigen showed suppression of established tumor growth but the efficacy was depending on the size. Moreover, in human PBMC studies, the bigger antigen showed better CD4 and CD8 T cell responses than the others. These data demonstrate that if the oncogenic potential of antigen could be eliminated, the biggest form of antigen would show better therapeutic effects.
Measuring fg/mL Concentrations of Cytokines in Cerebrospinal Fluid (CSF)
Authors: Georlette Daphne , Suschak Jessica , Aghvanyan Anahit , Navaratnam Manjula , Lockney Dustin , Biesso Arianna , Gardner Louis , Bartlett Dan , Porter Erica , Tamot Banita , Shukla Animesh , Miller Thomas , Mathew Anu , Wheeler Sarah E , Stengelin Martin , Wohlstadter Jacob N 
Published: January 2023

Abstract
Measuring low-abundance cytokines in CSF may be useful when studying brain and nervous system disorders. We recently developed novel immunoassays with fg/mL and sub-fg/mL detection limits that enable quantitation of low-abundance biomarkers in CSF. S-PLEXSM assays, utilizing MSD’s MULTI-ARRAY® electrochemiluminescence technology, were developed and analytically characterized and the concentrations of cytokines in CSF were measured using these assays.
Assays for the following cytokines were used: IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-17A, IL-21, IFN-γ, GM-CSF, TSLP, and TNF-α. The lower limits of detection achieved for S-PLEX assays for these analytes ranged from ~0.2 fg/mL to ~50 fg/mL. Spike recovery and dilution linearity were between 80% and 120%. Depletion experiments demonstrated specificity for each analyte. Cytokine levels in the majority of apparently healthy CSF samples were measurable for 8 out of the 12 analytes tested.
We have developed a next-generation assay format that is 100 to 1,000 times more sensitive than the current limits of standard ELISA technology. This increased sensitivity enables accurate quantitation of cytokine levels in the CSF of apparently healthy donors. These assays will allow for a better understanding of the role these cytokines play in brain and nervous system disorders.