bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of LPS Tolerance in Macrophages (48.2)
Authors: O'Carroll Christine , Fagan Ailis , Carmody Ruaidhri 
Published: January 2023

Abstract
Lipopolysaccharide (LPS) tolerance is an essential immune-homeostatic response which prevents unnecessary over-activation of the inflammatory response. LPS tolerance induces a state of reduced responsiveness to repeated challenges with LPS, resulting in decreased expression of pro-inflammatory modulators, up-regulation of antimicrobials and other mediators promoting the resolution of inflammation. LPS tolerance is a transient state and LPS tolerant macrophages (MΦT) return to an LPS responsive state within days of initial stimulation. Macrophages which have recovered from a tolerant state (MΦR) remain largely uncharacterised. Gene expression profiling reveals that LPS tolerance recovery triggers an extensive modulation of the gene expression profile of macrophages distinct from that of (MΦT) and LPS activated macrophages (MΦA). Recovery from LPS tolerance at 88 hours post treatment leads to expression of key pro-inflammatory genes at levels comparable to MΦA. MΦR show increased expression of key modulators of the immune response not previously observed during classical activation. We propose that MΦR represent a novel macrophage phenotype. Furthermore we propose that the recovery from LPS tolerance promotes a global modulation of gene expression with distinct expression profiles between genes promoting inflammation and genes key to the resolution of inflammation. In addition we present an expression analysis of NF-κB and its regulators in naïve, MΦT and MΦR cells.
P-selectin Glycoprotein Ligand-1 Expression on T cells is Required for Optimal Development of T Cell Memory in Response to Influenza Virus Infection (49.22)
Authors: Tinoco Roberto , Baaten Bas , Douglas Mia , Bradley Linda 
Published: January 2023

Abstract
Influenza viruses represent a major global health concern as they have the potential to cause worldwide pandemics characterized by increased morbidity and mortality. T cells are critical for eliminating virus-infected cells and their localization to the respiratory tract is crucial to resolving infection. The underlying mechanisms regulating effector and memory T cell responses after influenza virus infection remain unclear. Using a mouse model, we demonstrated that the adhesion molecule P-selectin is induced on lung endothelium after infection. Virus-specific T cells acquired the ability to bind P-selectin through expression and glycosylation of its ligand PSGL1. P-selectin binding antigen-specific T cells produced increased levels of IFN-γ and granzyme B in the lung when compared to non-binding T cells. Furthermore, P-selectin binding antigen-specific T cells persisted as memory cells, expressed IL-7 receptor, and localized primarily in the lung and its draining lymph node. PSGL1 knockout (KO) T cells, which do not bind P-selectin, expanded to a greater extent than wildtype T cells during primary infection. However, fewer PSGL1 KO T cells persisted following contraction and there was a profound defect in PSGL1 KO memory cell re-expansion. Our findings highlight a critical role for the P-selectin-PSGL-1 interaction in regulating local T cell responses. Modulating this interaction may provide a novel way to boost T cell memory and immunity to influenza viruses.
Nasopharyngeal colonization by Streptococcus pneumoniae elicits long-lasting immune responses that confer protection from invasive lung infections. (49.23)
Authors: Maung Nang , Goodwin Aaron , Munoz-Elias Ernesto , Clark Stacie , Camilli Andrew , Gerstein Rachel , Leong John 
Published: January 2023

Abstract
Streptococcus pneumoniae can cause life-threatening pulmonary and systemic infection, but more commonly resides harmlessly in the nasopharynx. We demonstrated that nasopharyngeal (n.p.) colonization by S. pneumoniae in mice generates an antibody response that provides at least 10-week protection against lethal lung challenge. B cells were required for this protection, and transfer of sera from n.p.-colonized mice or antibodies against phosphorylcholine, a common S. pneumoniae surface antigen, protected naïve mice. Whereas T-independent anti-capsular antibodies were previously shown to be sufficient for protection against bloodstream infection , we found that T cells were required to generate a fully protective response following n.p. colonization. Depletion of CD4+ T cells at the time of pulmonary challenge or colonization of MHC class II-deficient mice did not compromise protection, suggesting a limited role for CD4+ T cells in our model. Sub-lethal irradiation of mice, which inactivates memory B cells but not long-lived plasma cells, did not significantly diminish protection, and transfer of bone marrow cells enriched for long-lived plasma cells from n.p.-colonized mice to B cell-deficient recipients increased pneumococcal-specific antibody titers and afforded significant protection. Our results suggest that n.p. colonization is capable of eliciting extended protection against invasive pneumococcal disease by generating long-lived antibody-secreting cells.
Th17 cells mediate clade specific mucosal immunity (49.26)
Authors: Chen Kong , McAleer Jeremy , Weaver Casey , Kolls Jay , Paterson David 
Published: January 2023

Abstract
The IL-17 family of cytokines phylogenetically pre-dates the evolution of T-cells in jawed vertebrates, suggesting that the ontogeny of the Th17 lineage must have arisen to confer an evolutionary advantage to the host over innate sources of IL-17. Using a model of mucosal immunization with the encapsulated bacteria, Klebsiella pneumoniae, we show that intranasal immunization strongly induces IL-17 producing cells in the lung. Further analysis reveals that these cells are conventional Th17 cells and are antigen specific as they proliferate and produce IL-17 upon re-stimulation with heat-killed K. pneumoniae but not heat-killed Escherichia coli or Staphylococcus aureus. In vivo, immunization induced Th17 cells but not Th1 cells undergo activation/expansion and contraction overtime and the Th17 responses together with anit-K. pneumoniae antibody responses lead to the protection of immunized mice from live K. pneumoniae infection. However, B-cell/antibody mediated protection is limited only to the vaccine strain. In contrast memory Th17 cells proliferate in response to conserved outer membrane proteins and confer protection against subsequent bacterial challenge with several serotypes of K. pneumoniae, including the recently described multi-drug resistant New Delhi metallolactamase strain. These data suggest that Th17 adjuvants can improve vaccination strategies against drug resistant encapsulated bacteria.
Potent anti-inflammatory activity of a novel of class Hsp90 inhibitors for multiple sclerosis (51.1)
Authors: Zhou Dan , Liu Yuan , Ye Josephine , Ying Weiwen , Chimmanamada Dinesh , Shin Ogawa Luisa , Inoue Takayo , Rao Patricia , Wada Yumiko 
Published: January 2023

Abstract
Current therapies for multiple sclerosis (MS) generally reduce the frequency, severity and duration of relapses, and may slow disease progression, but are not curative. Clinical benefit also comes with side effects that range from mild to life-threatening. Heat shock protein 90 (Hsp90) is a molecular chaperone required for the stability and function of many proteins that modulate cellular response to environmental stresses. Due to its central role in multiple immune-mediated mechanisms, Hsp90 has recently emerged as a potential target for MS therapy. A number of the inflammatory mediators underlying MS are known Hsp90 client proteins, and Hsp90 blockade represents an attractive multi-targeted therapeutic approach given its alternative mechanisms of neuroprotection and inhibition of inflammation. Here we describe a novel class of small molecule inhibitors that preferentially target Hsp90 associated with immune-mediated pathways and which demonstrate potent inhibitory activity, high oral bioavailability and prolonged CNS exposure. Further, the lead compound displays broad activity in a number of relevant in vitro and in vivo models; including inhibition of cytokine production by human PBMCs, decreased nitric oxide production by primary rat astrocytes, and efficacy in an adoptive transfer model of MS. Taken together, this class of Hsp90 inhibitors may potentially provide new avenues to counter immune overstimulation in MS patients.
Comparison of protein expression by different lentiviral vectors (51.12)
Authors: Wang Nan , Rajasekaran Narendiran , Hou Tieying , Mellins Elizabeth 
Published: January 2023

Abstract
HIV1-derived lentiviral vectors have been widely used as gene delivery tools due to their potent transduction capacity and stable expression after chromosomal integration in dividing and non-dividing mammalian cells. Lentiviral vectors were screened for expressing the murine class II chaperone, invariant chain (Ii), in hematopoietic stem and progenitor cells (HSPC). We compared various lentiviral vectors using GFP as a reporter gene in 293T cells and HSPC. We assessed a dual promoter vector (DP) with separate promoters for Ii and GFP, a fusion protein vector (FU) that expresses Ii and GFP together under a single promoter, and T2A vector (T2A) in which Ii and GFP are separated by a self-cleaving 2A peptide. Similar percentages of 293T cells were positive for Ii expression after being directly transfected with DP and T2A. The percentage was slightly higher in FU transfected cells. 293T cells transfected with DP have the highest GFP expression. We packaged each vector into lentivirus and found that both Ii and GFP expression levels dropped in 293T cells. Further reduction in expression was seen in viral transduced HSPC. The trend toward different expression levels of Ii and GFP persisted whether lentiviral transduction or vector transfection was used. Only DP and T2A constructs, but not FU constructs, resulted in functional Ii expression. Based on these results, we hypothesize that the poor expression of Ii may be due to deleterious effects of Ii on assembly of competent virus.
Effect of targeted drug delivery with G5-MTX dendrimers on induced T regulatory (iTreg) CD4+ T cells in an inflammatory bowel disease (IBD) colitis model (52.1)
Authors: Belyakov Igor , Zimmermann Ellen , Isakov Dmitry , Fisk Elizabeth , Orr Nicholas , Schmiedlin-Ren Lee , Choi Seok Ki , Baker James 
Published: January 2023

Abstract
Inflammatory bowel disease (IBD) are complex immune mediated diseases involving the interactions of many cell types including the intestinal mucosal epithelia, macrophages, T-lymphocytes (Foxp3+ Treg, Th17, Th2) and dendritic cell subsets. Induced Treg (iTreg) play an essential role in immune tolerance in the control of chronic inflammation. Current approaches to IBD therapy utilize immunosuppressive medications including methotrexate (MTX), which is hepatotoxic. Targeted drug delivery of MTX for IBD treatment may be achieved by conjugating MTX to G5 PAMAM dendrimer nanoparticles. We investigated the effect of G5-MTX12 on inflammatory responses in dextran sulfate sodium (DSS) induced IBD in BALB/c mice. We evaluated the frequency of iTreg (Foxp3+Helios- CD4+ TCRαβ+ T cells) in mesenteric lymph nodes (MLN). We found that treatment with G5-MTX induces protection of mice against DSS colitis. We found that the weight of mice treated with G5-MTX was not significantly different from naïve animals. There was a 4.7-fold increase in frequency of iTreg in mesenteric lymph nodes in G5-MTX treated mice compared to naïve animals. Thus, we achieved a therapeutic effect with targeted G5-MTX in the IBD model in mice. This targeted drug effect may be mediated by iTreg that developed under sub-immunogenic antigen presentation during intestinal inflammation. Our data may suggest that a targeted drug delivery strategy with dendrimer technology may be effective with potentially less toxicity
Selenoprotein K is a novel target of m-calpain, and cleavage is regulated by toll-like receptor-induced calpastatin in macrophages (54.22)
Authors: Rose Aaron , Zhi Huang , Hoffmann Fukun , Norton Robert , Hoffamann Peter 
Published: January 2023

Abstract
Calpains are calcium (Ca2+)-activated cysteine proteases that modulate cellular function through cleavage of a variety of intra-cellular targets. Two major isoforms of calpain, u- and m-calpain, are expressed in different tissues and cell-types. Selenoprotein K (SelK) is a selenocysteine containing transmembrane ER protein involved in Ca2+ flux, and western blot analyses revealed one SelK band in mouse lymphocytes (both B and T cells) and two bands for SelK in mouse myeloid cells (macrophages, dendritic cells, and neutrophils). This led us to investigate cleavage of SelK and we found that m-calpain cleaved SelK in myeloid cells. The calpain-mediated cleavage of SelK occured between Arg81 and Gly82 resulting in the removal of the selenocysteine-containing portion of the cytosolic region of the protein. We have further linked this event to the activation state of macrophages. Resting macrophages were found to predominantly contain cleaved SelK while macrophages activated through toll-like receptors contained full length SelK. This was due to increased levels of calpastatin, which is the endogenous inhibitor of calpains. The identification of SelK as a novel target for calpain cleavage activity reveals a novel mechanism by which macrophage activation is regulated and studies are on-going to further characterize the role of calpain/calpastatin in macrophages.
Decreased expression of B- and T-lymphocyte attenuator (BTLA) is associated with progression of disease in idiopathic pulmonary fibrosis (IPF) (54.7)
Authors: Takahashi Stephenie , Blaine Kelly , Williams Jesse , Fu Yang-Xin , Noth Imre , Sperling Anne 
Published: January 2023

Abstract
IPF is an interstitial lung disease that affects over 128,000 people in the U.S., has a yearly mortality of 48,000, and has no effective treatment. Current theories on the etiology of IPF have not focused on the role of the immune system due to the inefficacy of steroid therapy. However, we have found in microarray studies that peripheral blood T cell co-stimulatory markers CD28, ICOS and BTLA, are associated and significantly predictive of early mortality in IPF patients. A significant association was found between decreased BTLA mRNA levels and decreased predicted diffusing capacity of carbon monoxide (DLCO) levels, a clinical measure of disease severity. Analysis of all samples demonstrated a significant and dramatically decreased BTLA surface expression on both peripheral CD4 and CD8 T cells, but not B cells, in rapidly progressing patients compared to controls or stable patients. Similar, but less dramatic decrease in surface expression is found for CD28 and ICOS. In a bleomycin-induced pulmonary fibrosis model, BTLA-/- mice, and to a lesser extent ICOS-/- mice, show increased numbers of lung CD4+ T cells and macrophages, significantly decreased weight, and more severe lung fibrosis when compared to controls. These results support the role of altered co-stimulatory molecules in the progression of IPF implying that these molecules are involved in the pathogenesis of disease progression.
Asthma: A missense SNP in the FCER2 (CD23) gene favors increased IgE production by human B cells (57.2)
Authors: Chan Marcia , Gigliotti Nicole , Rosenwasser Lanny 
Published: January 2023

Abstract
Asthma now is the most common chronic condition in children. Many asthmatics are unresponsive to therapy and genetic predisposition likely contributes. Pharmacogenetic studies have highlighted novel sequence variants in several genes. One such gene is fcϵr2, encoding the low affinity IgE receptor CD23 (FcϵRII). CD23 is expressed on several cell types and has several roles in IgE-mediated immunity, including antigen focusing and presentation to TH2 cells; mediating production of pro-inflammatory cytokines by monocytes; transporting IgE and allergen-IgE complexes through gastrointestinal epithelium; and participating in both positive- and negative-feedback mechanisms in regulation of IgE synthesis by B cells. Single nucleotide polymorphisms (SNPs) in fcϵr2 are associated with increased risk of exacerbations in asthmatics treated with inhaled corticosteroids but functional relevance of these SNPs in fcϵr2 remains unknown. One SNP (rs2228137) encodes a nonsynonymous amino acid change (R62W). We established stable human B cell transfectants expressing either wild type CD23b-R62 or CD23b-R62W. Stimulation through CD23 causes differences in ERK1/2 activation between the transfectants. Transfectants expressing CD23-R62W exhibited increased activation of the transcription factor Egr-1 (found in the IL-4R promoter region), and increased expression of IL-4R. These data suggest that the R62W SNP may favor increased IgE production through increased responsiveness to IL-4.
Development of mRNA signatures tools to detect in vivo basophil phenotypes: IL-3 signatures (57.5)
Authors: MacGlashan Donald 
Published: January 2023

Abstract
There are several well-recognized phenotypes of peripheral blood basophils (e.g., non-releasers) but the underlying bases for the phenotypes are not explained. IL-3 has many effects on basophil maturation and function and is known to induce considerable changes in basophil phenotype. Purified human basophils were cultured with or without IL-3 for 24 hours and the microarray expression profile, relative to pre-culture, analyzed for transcripts that acted in a reciprocal fashion depending on the condition. A 51- gene transcript set was identified that was composed 34 transcripts that increased with IL-3 and decreased without IL-3 and a further 17 transcripts that did the reverse. The changes were stable in time, with 3-day cultures generating a concordant signature. The inverted nature of the response to ±IL-3 suggested that the normal circulating basophil is in a sensitive balanced state of IL-3 exposure. For example, it could detect the early generation of IL-3 that follows IgE-mediated stimulation of basophils. When this signature tool was applied to freshly isolated basophils, it was observed that the signature was stable in time for the same subjects’ basophils. But when comparing the non-releaser and releaser phenotypes, the signature was consistent with a low-level deficiency in exposure to IL-3 or an IL-3-like cytokine in the “non-releaser phenotype. These studies identified an IL-3 signature to probe changes in basophils occurring in vivo.
Methods for quantification of In vitro cell-substrate contacts. (58.1)
Authors: Cammer Michael , Vardhana Santosh , Novak Hila , Sacristan Rock Catarina , Liapis Anastasia , Depoil David , Dustin Michael 
Published: January 2023

Abstract
A common problem in in vitro immunology research is characterizing cell spreading on substrates by morphology and by protein or other molecule distributions. Here we describe automated and semi-automated methods for measuring protein distributions at the cell-substrate interface which are key indicators of biological function. These methods may be applied to images from live or fixed cells. Using macros in ImageJ, we have developed methods to quantify the symmetry of attaching or spreading cells and the radial distributions of proteins. These distributions may be correlated with morphometry and provide insight into cell physiology. Examples in T Cells are shown regarding the relative localizations protein kinases under various treatments and of ICAM and TCR in a myosin mutant. In addition, a simple method is described to measure the movement of proteins in live cells where automated computer detection is not feasible.
The susceptibility of CXCR3KO mice to EAE is associated with the increased numbers of Th17 infiltrating in CNS (60.4)
Authors: Chung Chen-Yen , Liao Fang 
Published: January 2023

Abstract
CXCR3 and its ligands are highly associated with neuronal inflammation. Given that Th17 cells play a critical role in EAE pathogenesis and that Th17 cells have been shown to express CXCR3, we intended to investigate the effect of CXCR3 on EAE, particularly focusing on Th17 cells. In this study, we demonstrated that CXCR3KO mice developed more severe EAE as compared with wild-type mice. Of interest, CXCR3KO mice exhibited higher numbers of Th17 cells infiltrating in CNS as compared with wild-type mice. In accordance with this result, using the passive EAE model, we showed that mice receiving Th17 cells derived from MOG-immunized CXCR3KO mice developed more severe EAE as compared with mice receiving Th17 cells derived from MOG-immunized wild-type mice. Furthermore, intracellular staining revealed that CXCR3KO mice exhibited the increased numbers of Th17 cells expressing GM-CSF, a recently identified encephalitogenic cytokine that contributes to the effector phase of EAE. Altogether, we provide evidence that CXCR3 plays a protective role rather than a immunopathgenic role in EAE via negative regulation of Th17 cells infiltrating in CNS. The mechanism underlying the increased numbers of Th17 cells infiltrating in CNS of CXCR3KO mice leading to the susceptibility to EAE is currently under investigation.
Contribution of CD11c+ GR1+ cells to the initiation of lymph node vascular growth after immunization via the secretion of IL-1β (61.8)
Authors: Benahmed Fairouz , Tian Sha , Lu Theresa 
Published: January 2023

Abstract
Immune stimulation results in draining lymph node swelling and vascular-stromal growth. Lymph node endothelial cell proliferation is upregulated at day 2 after immunization and this early event is dependent on CD11c+ cells and VEGF. Fibroblast-type reticular stromal cells (FRC) are the principal VEGF-expressing cells within lymph nodes. Here, we test the hypothesis that CD11cmed MHCIImed GR1+ cells that accumulate in the draining lymph node plays a role during the upregulation of endothelial cell proliferation at day 2. We show that depletion of GR1+ cells induces a reduction in endothelial cell proliferation in comparison with control mice. In vitro, CD11c+ cells isolated from immunized lymph nodes are able to stimulate VEGF secretion by FRCs. CD11c+Gr1+ cells express IL-1β, and anti-IL-1β treatment reduces lymph node endothelial cell proliferation. Our results suggest that the CD11c+GR1+ cells contribute to the initiation of lymph node vascular growth via the secretion of IL-1β, which then induces increased VEGF expression by FRCs.
Ceramide deficiency in the epidermis leads to development of psoriasis-like lesions associated with IL-23-dependent proliferation of γδ-17 cells (62.1)
Authors: Nakajima Kimiko , Kataoka Sayo , Goto-Inoue Naoko , Terao Mika , Setou Mitsutoshi , Katayama Ichiro , Takeda Junji , Sano Shigetoshi 
Published: January 2023

Abstract
It has been recognized that ceramide levels are decreased in the epidermis of patients with atopic dermatitis and psoriasis. However, the underlying mechanism by which ceramide deficiency leads to skin inflammation still remains unclear. Here, we generated Sptlc2 targeted mice under control of the keratin 5 promoter (referred to as K5-SPT-KO mice), by which their keratinocytes were devoid of serine palmitoyltransferase (SPT), the rate-limiting enzyme for de novo sphingolipid synthesis. K5-SPT-KO mice have decreased levels of ceramide in the epidermis and these mice demonstrate barrier dysfunction. From 2 weeks of age, they develop skin inflammation, showing psoriasis-like histopathologic changes and inflammatory cell infiltrates in the dermis. While expression of IFN-γ was not observed in the inflamed skin and draining lymph nodes, increased levels of IL-17 and IL-22 were detected. Strikingly, K5-SPT-KO mice showed increased numbers of γδ-T cells that produce IL-17 (γδ-17) in the skin lesion and lymph nodes and most of them also produce IL-22, similar to Th17 cells. However, much fewer γδ-17 cells are detected in the skin and lymph nodes of wild-type mice. In vivo administration of anti-IL-12/23p40 antibody ameliorated the skin lesions and reduced the number of γδ-17 cells in K5-SPT-KO mice. Therefore, we conclude that ceramide deficiency in the epidermis results in the development of psoriasis-like lesions, likely mediated by IL-23-dependent γδ-17 cells in mice.