bluuebunny/crossref_metadata_2025_split huggingface dataset:
Serum soluble Siglec-8 in hypereosinophilic syndromes
Authors: Legrand Fanny , Cao Yun , Bochner Bruce S , Klion Amy D 
Published: January 2023

Abstract
Siglec-8, selectively expressed on eosinophils, basophils and mast cells, is the target of novel biologics in clinical development for eosinophil and mast cell-associated disorders. Since soluble Siglec-8 (sSiglec-8) can be detected in serum and could interfere with the efficacy of biologics targeting Siglec-8, the aim of the present study is to assess sSiglec-8 levels in a large cohort of subjects with hypereosinophilic syndrome (HES). sSiglec-8 levels were quantified in serum from normal (n=9) and eosinophilic (n=123) subjects using an in-house sandwich ELISA (limit of detection: 0.5 ng/mL) and correlated with Siglec-8 expression on the surface of eosinophils (measured by flow cytometry on whole blood eosinophils gated as granulocytes CD45+CD16neg) and absolute eosinophil count (AEC). The effects of treatment were also examined.
Serum levels of sSiglec-8 were undetectable in 28 subjects, did not differ between untreated HES subjects and normal donors (GM 1.23 and 1.53 ng/mL), and did not correlate either with AEC (n=123) or with Siglec-8 surface expression on blood eosinophils (n= 35). There was also no clear relationship between the clinical subtype of HES and levels of sSiglec-8. Levels measured before and after varied HES treatments in 7 subjects showed a significant decrease from 1.82 to 0.50 ng/mL (P&lt;0.05). In contrast, serum from 8 HES patients at two different time points on no treatment showed no significant change (GM 2.35 and 2.10 ng/mL respectively). These data suggest that, despite the lack of correlation with AEC, eosinophils are a significant source of sSiglec-8 in serum. Whether serum sSiglec-8 is predominantly from tissue eosinophils or mast cells, or requires eosinophil activation, is currently under investigation.
Adam8 limits emphysema development in smoke-exposed mice: A new player in the pathogenesis of COPD
Authors: Rojas Joselyn Joanna , Polverino Francesca , Gai Xiaoyan , Rout Amit , Wang Xiaoyun , Yambayev Ilyas , Owen Caroline Anne 
Published: January 2023

Abstract

Background
A disintegrin and metalloproteinase-8 (ADAM8) is expressed by lung cells implicated COPD pathogenesis. It’s not known whether ADAM8 contributes to COPD pathogenesis.

Methods
ADAM8 expression was measured in the lungs of COPD patients and controls, and mice exposed to cigarette smoke (CS) versus air by staining lung sections for ADAM8 and markers of different lung cells. Emphysema development and small airway fibrosis were evaluated wild-type (WT), Adam8−/− and Adam8 bone marrow chimeric mice exposed to air or CS for 6 months. Lung inflammation, alveolar septal cell apoptosis, and lung oxidative stress levels were measured in WT vs. Adam8−/− mice exposed acutely to air or CS.

Results
Adam8 expression is reduced in alveolar macrophages and lung epithelial cells in COPD patients and CS-exposed WT mice. CS-exposed Adam8−/− mice had greater airspace enlargement, alveolar septal cell apoptosis, and oxidative stress levels (but similar small airway fibrosis) versus CS-exposed WT mice. Both leukocyte- and lung parenchymal cell-derived Adam8 mediate Adam8’s protective activities in the lung. CS-exposed Adam8−/− mice had greater lung macrophage counts than WT mice. ADAM8 does not degrade mediators of inflammation or regulate monocyte adhesion or migration. Adam8−/− alveolar macrophages were protected from CS-induced activation of the intrinsic apoptosis pathway.

Conclusions
ADAM8 protects against CS-induced emphysema development by reducing lung inflammation by increasing macrophage apoptosis and reducing alveolar septal cell death. However, ADAM8’s expression is reduced in COPD lungs. ADAM8 may be a therapeutic target for COPD.
RELM protein-induced CXCR3 ligands in pulmonary hypertension
Authors: Van Raemdonck Katrien , Lin Qing , Fang Xia , Skinner John , Fan Chunling , Yamaji-Kegan Kazuyo , Johns Roger A. 
Published: January 2023

Abstract
In pulmonary hypertension (PH), inflammation and angiogenesis go hand in hand. Inflammatory mediators exert direct effects on endothelial (EC) and vascular smooth muscle cells (VSMC) and attract leukocytes to affected vascular beds. Resistin-like molecules (RELM) were previously implicated in vascular remodeling in PH. Structural and functional parallels can be drawn between chemokines and RELMs. Resistin (Retn) stimulates SMC proliferation and regulates EC adhesion and apoptosis. The less-studied Relm-β has similar effects. RELMs also modulate chemokine production. Our data suggest that Relm-β contributes to pathogenic upregulation of CXCL10 and CXCL11, both IFNγ-inducible CXCR3 ligands. These are known players in angiogenic disorders and have been associated with PH. Under hypoxia, mice did exhibit increased pulmonary expression of CXCL11 in a pilot study. In vitro chemokine levels indicate that RELMs stimulate CXCL10 and CXCL11 release by activated monocytic THP1 cells. Because chemokines may locally affect VSMC proliferation, cause EC apoptosis, and attract T cells that exacerbate inflammation, we hypothesize that CXCR3-driven vascular remodeling contributes to the pathogenic role of RELMs in PH. While the receptor remains unknown, our preliminary data suggest Relm-β-induced inhibition of PPARγ activity to regulate chemokine release. Our endeavors continue to unravel the receptor and downstream pathways involved. As development of RELM-targeted therapies progresses, understanding the full range of biological activities of, not only Retn, but also Relm-β, remains crucial. CXCR3 ligands have the potential to serve as valuable biomarkers as we evaluate the efficiency of innovative therapies in the field of PH.
Using MHC Binding Assays to Investigate HLA-Dependent Drug Directed Immune Responses
Authors: Kaever Thomas , Metushi Imir G , Pavlos Rebecca , Sidney John , Ostrov David A , Buus Soren , Redwood Alec , Sette Alessandro , Hunt Donald F , Phillips Elizabeth J , Peters Bjoern 
Published: January 2023

Abstract
Immune mediated adverse drug reactions (IM-ADRs) pose a significant risk to human health, and have been shown to be linked to specific human leukocyte antigen (HLA) alleles. Hypersensitivity syndrome induced by abacavir, the strongest such genetic association found to date, is driven by drug-specific activation of cytokine producing, cytotoxic CD8+ T cells exclusively found in individuals expressing the HLA-B*57:01 allele variant. We have previously shown that abacavir interacts with the peptide binding cleft of the HLA-B*57:01 molecule, thus increasing the affinity of certain self-peptides generating an altered self-peptide repertoire. In MHC binding studies a second drug, acyclovir, also revealed a dose dependent increase in affinity for peptides that contain small, hydrophobic residues at the C terminus. Peptide elution studies performed in the presence of acyclovir showed an elevated number of endogenously bound peptides with a C-terminal isoleucine. Accordingly, we concluded that acyclovir acts by the same mechanism as abacavir, albeit at a lesser magnitude. Our data showed a maximum increase of affinity of 2.5-fold for acyclovir, when the effect of abacavir was as high as 1000-fold. Acyclovir is not known to cause IM-ADRs. We concluded that the moderate effect of acyclovir on binding affinity was not sufficient to cause a hypersensitivity reaction. Here we applied the same screening strategies to four compounds that are involved in IM-ADRs with strong known HLA association, to determine if drug specific effects of these molecules could be observed. While no effect as striking as that for abacavir could be detected, several notable changes in repertoire were associated with the presence of these compounds.
High output flow cytometry array profiling facilitates discriminant phenotyping of peripheral blood cells from patients with different forms of autoimmune uveitis
Authors: Nowatzky Johannes , Manasson Julia , Resnick Ezra , Stagnar Cristy , Manches Olivier 
Published: January 2023

Abstract
The information content of multi-parametric flow cytometry-based immune-phenotyping experiments is often underexploited given the paucity of adequate tools and strategies for large-scale unbiased data analysis. Here, we developed and applied a data analysis approach taking into account all mathematically possible combinations of markers in a given flow cytometry panel. We analyzed flow cytometry data generated from peripheral blood samples of healthy humans and patients with autoimmune uveitis in the setting of 2 different autoimmune diseases: Behcet’s disease and sarcoidosis. Original FACS data files were mined from Dryad Digital Repository http://dx.doi.org/10.5061/dryad.v6ste with reference to http://dx.doi.org/10.1371/journal.pcbi.1003215, and gated in bivariate plots. We employed combinatory mathematics to generate a matrix quantifying the representation of all possible cell populations using a given set of markers within the respective starting population and coded the algorithm to create a Java-based platform enabling the computation of large numbers of markers. Resulting metadata were visualized in a heat map approach. Our method enabled clustering of healthy vs diseased subjects using only 4 common markers (CD3, C8, CD197, and CD45), and of sarcoid vs Behcet’s patients with minimal error. Our approach demonstrates that multi-dimensional analysis of flow cytometry data allows meaningful large-scale screening of biologically relevant markers enabling classification and characterization of states of health and autoimmune disease. The approach is unbiased and has the potential to facilitate the discovery of cell populations with relevance as potential biomarkers or biological research targets.
Bacterial Dysbiosis Associates with Functional Intraepithelial Lymphocyte Changes in Inflammatory Bowel Disease and Spondyloarthritis
Authors: Ohri Neha , Gerich Mark E , Fennimore Blair P , Ir Diana , Robertson Charles E , Regener Emilie , Caplan Liron , Wagner Brandie D , Frank Daniel N , Kuhn Kristi 
Published: January 2023

Abstract
Dysbiosis occurs in spondyloarthritis (SpA) and inflammatory bowel disease (IBD), subdivided into Crohn’s Disease (CD) and Ulcerative Colitis (UC). The immunologic consequences of dysbiosis have not been defined. Intraepithelial lymphocytes (IELs) are T cells within the intestinal epithelium that are in close contact with bacteria, and as such, are likely to be modulated by dysbiosis. We correlated IELs with resident bacteria in SpA, IBD, and controls. Subjects with biopsy-proven IBD (N=10 with CD and 7 with UC), SpA fulfilling ASAS criteria (N=5), and healthy controls (N=15) were evaluated for fecal microbiome by 16S rRNA sequencing and IELs from colon biopsies analyzed by flow cytometry and ELSIA. Subjects with SpA had significantly lower numbers of IELs compared to controls (p=0.03). Subjects with CD had significantly increased IL-17A, (p=0.03) and IFN-γ (p&lt;0.01) whereas those with UC had higher IL-1β (p=0.01) compared to controls. Both subjects with CD and SpA had significantly increased secretion of TNF-α (p=0.04). Correlating cytokines to bacteria revealed an association between TNF-α and the Simpson Diversity Index in subjects with UC (Spearman’s r=0.943, p&lt;0.01); in subjects with CD, Fusobacterium had a negative correlation with TNFα+IFNγ+IL-1β (Spearman’s r=−0.786, p=0.02). Sequencing results from subjects with SpA are pending. Our data indicate differences in IEL function among subjects with SpA, CD, and UC compared to healthy controls. We hypothesize that the correlations between dysbiosis and IEL function are relevant to the pathogenesis of SpA and IBD. Future studies will be aimed at further understanding mucosal T cell and microbial interactions and may offer new therapeutic targets in these diseases.
Expression and regulation of chemokine receptor CXCR3 (CD183) on human gastrointestinal stem cells (hGISC) derived primary epithelial cells
Authors: Panja Asit B , Panja Prerona , Bhatt Rachana , Joseph Ansamma 
Published: January 2023

Abstract
Expression of Chemokine receptor CXCR3 (CD183) by cells in inflamed tissues play essential role in the development of chronic conditions by attracting leucocytes to the inflammatory sites. CXCR3 expression in intestinal epithelium has also been implicated in celiac disease by means of its potential role in peptide (gliadin) uptake and triggering of immune responses in the small intestine. Therefore, having an CXCR3 expressing intestinal epithelial cell system in long-term culture would be a valuable tool for discovery and development of new treatments for celiac and/or other gastrointestinal inflammatory diseases. Earlier, we have developed a method for growing human intestinal primary epithelial cells (HIPEC) from isolated human gastrointestinal stem cells (hGICSs) in long-term culture. Now we examined two small intestinal HIPEC lines for CXCR3 expression by immunocytochemical staining and subsequent fluorescence microscopic analysis.
Our results showed that CXCR3 was either undetectable or weakly expressed in normal jejunal HIPECs that had columnar characteristics whereas CXCR3 was detected on HIPECs with enriched MUC2 positive goblet epithelial cells. CXCR3 was also detectable in HIPECs derived from an actively inflamed tissue from a patient with ulcerative colitis (UC). Stimulation of normal HIPECs with proinflammatory signaling agents LPS, TNFα or IFNγ showed a minimum effect on the level of CXCR3 expression when added individually. However combinations of LPS with either TNFa or IFNg or both increased the CXCR3 expression on HIPECs.
These findings may help better understand of the disease mechanisms and development of new treatment for inflammatory conditions such as celiac disease and/or ulcerative colitis.
Skin inflammation induced by lupus serum was inhibited in IL-1R deficient mice
Authors: Deng Guo-Min , Li Xiaoyan , Guo Xuanxuan , Liu Huicheng , Gao Gongmin , Xu Guangqiong , Fei Xibin , Fang Xiang , Qiao Wei 
Published: January 2023

Abstract
Skin lesion is the second common clinical manifestations of systemic lupus erythematosus (SLE), but its pathogenesis is not clear. IL-1 is a proinflammatory cytokine, and its role in SLE skin lesion is still unclear. We used IL-1R deficient mice and other gene deficient mice to study the role of IL-1 in the lupus serum -induced skin inflammation. We found that the severity of skin inflammation induced by lupus serum was significantly reduced in IL-1R deficient mice and caspase-1 deficient mice. IL-1R deficiency did not affect the expression of FcγRI (CD64), FcγRII (CD32) and MHC class II (CD74) induced by lupus serum. IL-1R deficiency also reduced the lipid raft clustering, and decreased expression of MCP-1 and TNFa in monocytes. Skin inflammation and keratinocyte proliferation were significantly decreased in TNFa deficient mice. Our findings indicate that IL-1 plays an important role in skin lesions of SLE. This study suggests that IL-1 is a therapeutic target in skin lesions of SLE.
The chromatin organizer SATB1 governs the epigenetic repression of the co-inhibitory receptor PD-1 in T cells
Authors: Payne Kyle K , Stephen Tom L , Chaurio Ricardo A , Allegrezza Michael J , Zhu Hengrui , Perez-Sanz Jairo , Perales-Puchalt Alfredo , Nguyen Jenny M , Vara-Ailor Ana , Eruslanov Evgeniy B , Borowsky Mark E , Zhang Rugang , Laufer Terri M , Conejo-Garcia Jose R 
Published: January 2023

Abstract
Despite the importance of Programmed Cell Death-1 (PDCD1/PD-1) in inhibiting T-cell effector activity, the mechanisms regulating its expression in anti-tumor lymphocytes remain poorly defined. Here we show that the chromatin organizer Special AT-rich Sequence-Binding Protein-1 (Satb1) is required to restrain activation-induced PD-1 expression. We demonstrate that, mechanistically, Satb1 physically interacts with a nucleosome remodeling deacetylase (NuRD) complex, which it recruits to Pdcd1 regulatory regions. This molecular complex thus drives histone de-acetylation and results in PD-1 repression in T cells. Accordingly, T-cell-specific Satb1 deficiency results in a 40-fold increase in PD-1 expression. Intriguingly, tumor-derived Transforming growth factor (Tgf)-β decreases Satb1 expression in T cells through binding of Smad Family Member (Smad) proteins to the Satb1 promoter, while Smad also competes with Satb1/NuRD for binding to Pdcd1 enhancers, cooperatively unleashing PD-1 expression in a Satb1-dependent manner. Consequently, Satb1-deficient tumor-reactive T cells lose their effector activity more rapidly than wild-type T cells at PD-L1+ tumor beds, but these differences are abrogated by sustained PD-L1 blockade. Therefore, we demonstrate that Satb1 is an epigenetic controller of PD-1 expression, and that Tgf-β signaling contributes to T cell dysfunction within the tumor microenvironment by inhibiting Satb1-mediated repression of PD-1.
Resistance to PD1 blockade in the absence of metalloprotease-mediated LAG3 shedding
Authors: Andrews Lawrence Peter , Moskovitz Jessica , Szymczak-Workman Andrea L , Bruno Tullia C , Workman Creg J , Vignali Dario AA 
Published: January 2023

Abstract
Inhibitory receptors control immune responses preventing exacerbated T cell activation and limiting antitumor immunity. LAG3 co-expression with PD1 phenotypically marks functionally exhausted tumor-specific T cells, however LAG3 expression and function is itself regulated by ADAM metalloproteinase-mediated cell surface cleavage. To investigate the impact of LAG3 shedding on T cells within tumors, a conditional knock-in mouse was generated that results in a non-cleavable form of LAG3 (LAG3.NC). LAG3.NC CD4Cre, LAG3.NC ThPOKCreERT2 and LAG3.NC E8ICre mice (restricting LAG3.NC to all T cells, CD4+ T cells and CD8+ T cells, respectively) exhibit enhanced LAG3 expression on the respective T cell subsets infiltrating MC38 tumors. Upon therapeutic administration of anti-PD1, MC38 tumor-bearing wild-type mice show significant tumor regression and 40% become tumor-free, resulting in long-term survival. LAG3.NC CD4Cre and LAG3.NC ThPOKCreERT2 resist anti-PD1 therapy and succumb to tumor growth. However, this phenotype is not observed in LAG3.NC E8ICre mice, resulting in a similar frequency of tumor-free mice to controls. CD8+ TIL isolated from LAG3.NC CD4Cre mice do not show enhanced IFN-g/TNF-a production or proliferation as observed in controls or LAG3.NC E8ICremice following anti-PD1. Failure to mediate an antitumor immune response by LAG3.NC CD4Cre, but not LAG3.NC E8ICre mice, suggests that LAG3 cleavage on CD4+ T cells, including Tregs, is necessary for CD8+ TIL to elicit an effective immune response with anti-PD1. Selective ADAM10 inhibition in vivo, preventing LAG3 shedding, also reduces cytokine release, proposing that metalloproteinase-mediated LAG3 cleavage limits the efficacy of the antitumor immune response.
Staphylococcus aureus alpha-toxin and Clumping Factor A are pathogenic and immunotherapeutic targets against a hematogenous implant-related biofilm infection
Authors: WANG YU , Cheng Lily , Helfer David R , Ashbaugh Alyssa G , Miller Robert J , Tzomides Alexander J , Thompson John M , Ortines Roger V , Tsai Andrew S , Liu Haiyun , Dillen Carly , Archer Nathan , Cohen Taylor S , Tkaczyk Christine , Sellman Bret R , Miller Lloyd S 
Published: January 2023

Abstract
Infection is a major impediment to the long-term success of implantable medical devices. Treatment of these infections is complicated by bacteria biofilms, which form on the implants and block penetration of immune cells and antibiotics. Hematogenous implant-related infections following a transient bacteremia are particularly problematic because they can occur at any time in a previously stable implant. To evaluate alternative targeted immune-based therapies against these infections, we developed a hematogenous infection model in which an orthopaedic titanium implant was surgically placed in the legs of mice followed by an intravenous injection of Staphylococcus aureus 21 days later. This resulted in a marked propensity for a hematogenous implant-related infection comprised of septic arthritis, osteomyelitis with neutrophil abscess formation in the bone and biofilm formation on the implants in the surgical legs compared with sham surgical legs without implant placement or contralateral nonsurgical normal legs. Prophylaxis with two neutralizing human monoclonal antibodies directed against S. aureus virulence factors, secreted alpha-toxin (AT) and surface expressed clumping factor A (ClfA) inhibited biofilm formation in vitro and the hematogenous implant-related infection in vivo. Our findings suggest that AT and ClfA are important pathogenic factors that could be targeted as a novel immunotherapeutic against S. aureus hematogenous implant-related infections.
Bone marrow T cells populations are differently affected by sepsis than their splenic counterparts in the murine sepsis cecal ligation and puncture (CLP) model
Authors: Skirecki Tomasz , Swacha Patrycja , Hoser Grażyna , Kozłowska Ewa 
Published: January 2023

Abstract
Sepsis is known to induce lymphopenia which correlates with secondary infections and mortality. In sepsis survivors the lymphocyte count restores with time, although the T cell responses are impaired. So far, the mechanisms of T cells recovery are not fully understood and the murine studies were focused on the peripheral lymphatic organs in which the spleen was most common. Our study was designed to elucidate the recovery of bone marrow (BM) T cells which contain population of memory cells, after the CLP murine sepsis. In our model, mild CLP produced peripheral lymphopenia which was lowest at 48h after CLP but after 7 days circulating T cells count was rebounding. 7 days after CLP the BM frequencies of naïve and central memory (CD44+CD62L+) CD4 cells were unchanged while they were reduced in the spleen. Percentage of effector memory (CD44+CD62L−) CD4 and CD8 cells increased in the BM and at the same time they dramatically decreased in the spleen. On the contrary, the effector (CD44−CD62L+) CD4 and CD8 cells expanded at that time point in the spleen while they did not change in the BM. At D7 after CLP, the rates of BrdU+ proliferating CD4+ and CD8+ cells were markedly higher in the BM than spleen. Among the BM CD4+ cells most of the proliferating cells were the effector memory cells, while the effector CD4+ had highest proliferation rate in the spleen. Similarly, CD8 effector memory cells in the BM and CD8 effector cells were most extensively proliferating in the BM and the spleen, respectively. Taken together, our data point to the BM as the place of effector memory T cell proliferation during post-sepsis recovery.