bluuebunny/crossref_metadata_2025_split huggingface dataset:
CCR2 identifies two subtypes of pathogenic human type 17 Th cells and has a non-redundant role in their transendothelial migration
Authors: Parween Farhat , Singh Satya P , Kathuria Noshin , Zhang Hongwei , Farber Joshua M 
Published: January 2023

Abstract
We have shown previously that CCR2 is expressed on highly differentiated human Th memory cells as well as mucosal associated invariant T (MAIT) cells. CCR2+ T cells typically co-express a combination of other inflammation-associated chemokine receptors, including CCR6, which is found on all type 17 T cells. We found that the CCR6+ CCR2+ human Th cell population is enriched in cells with a pathogenic cytokine and cytotoxic profile. Single-cell analysis revealed two subtypes of such CCR6+ CCR2+ cells, either biased to produce GM-CSF or IFNg. Pathogenic activity requires migration into inflamed tissue, and to understand the roles for CCR2, CCR6 and other chemokine receptors expressed on these cells in extravasation we used flow chambers containing HUVEC activated with TNFa and/or IFNg. We found that although CCR6, CCR5 and CXCR3 (but not CCR2) were capable of mediating T cell arrest, only CCR2 was active in transendothelial migration. CCR2’s activity was a function of the localization of its principal ligand, CCL2, in endothelial cells. Native CCL2 was found intracellularly but not on the cell surface. Immediate pre-treatment of cells with CCL2 or using HUVEC transduced to express a CCL2-CXCL9 chimeric chemokine that bound to surface glycosaminoglycans enabled CCR2 to mediate T cell arrest. These studies reveal the integration of transendothelial trafficking and a pathogenic effector profile in type 17 human Th cells and demonstrate redundant and non-redundant roles for the multiple chemokine receptors on these cells due to chemokine positioning. Our findings could inform more effective combinatorial inhibition or expression of chemokine receptors/ligands to regulate the migration of highly inflammatory T cells.
Supported by NIH
Role of paracrine mTOR signaling in regulating thymus size and function.
Authors: Wedemeyer Sarah A , Jones Nicholas , Emtage Simon , Griffith Ann 
Published: January 2023

Abstract
The thymus is the primary site of T cell generation, and the unique thymic stromal microenvironment directs T cell differentiation, self-tolerance and self-restriction. However, the size of the thymus declines precipitously relatively early in life, resulting in declining production of naïve T cells. Homeostatic mechanisms drive a shift toward an oligoclonal T cell memory, leaving the elderly less responsive to vaccines and new infections, especially viral infections. Preventing or reversing age-associated thymic atrophy therefore holds potential for extending the healthspan. Mechanisms governing thymic atrophy have been difficult to identify because the primary targets of atrophy, cortical thymic stromal cells, are rare and difficult to isolate. Using an informatic approach to characterize the transcriptional response of thymic stromal cells during age-related atrophy or experimental regeneration, we previously showed that cortical thymic epithelial cells (cTECs) display a unique morphology characterized by extensive looping projections. Atrophy is associated with contraction of these projections, which are renewed during induced regeneration. Further informatic analysis suggested that paracrine mTOR signaling in cTECs promoted by ligands expressed in medullary TECs (mTECs) may be required to maintain the cellular architecture of the cortical stroma and preserve thymus size. To test these hypotheses, we generated two new tissue-specific transgenic mouse lines overexpressing mTOR-activating ligands (IGF1 and FGF21) in most mTECs. Ongoing experiments aim to characterize the impact of increased paracrine mTOR signaling on age-associated thymic dysfunction. This work was supported by N.I.H. R21AI154109.
Supported by grant from NIH (R21 AI154109)
Risky business blockade: RAG2 basic region blocks V(D)J recombinase function upon genotoxic stress in DNA damage response
Authors: Simpson Destiny A , Rodgers Karla K 
Published: January 2023

Abstract
The V(D)J recombinase, composed of the recombination activating gene (RAG) proteins, RAG1 and RAG2, generate functional antigen receptor genes through the repeated generation of DNA double strand breaks and DNA repair. However, this process can lead to increased genomic instability particularly in the presence of excess DNA breaks. Previously, we showed in pre-B cells that full length RAG2, a nuclear protein, will export to the cytoplasm with DNA damage in an ATM-dependent manner with eventual recovery back into the nucleus following DNA repair. We propose that re-localization of RAG2 prevents RAG-mediated DNA cleavage in the presence of excess DNA ends. Here, by using fluorescently tagged RAG proteins, we show that DNA damage-induced export of the V(D)J recombinase can be recapitulated in non-lymphoid cells. Further, we determined that mutations in the intrinsically-disordered region of RAG2 have distinct effects on this process. Specifically, a basic region mutant of RAG2 (K/R 518–524 A) alters the localization pattern of the V(D)J recombinase, and yet its pre-DNA damage localization pattern is re-established upon DNA repair. Conversely, a T490A mutant blocks relocalization of RAG2 following genotoxic stress for both the single T490A mutant and the combined T490A/basic region mutant. Together, these results demonstrate the role of the intrinsically-disordered region in RAG2 in regulating the mobility and subcellular localization of the V(D)J recombinase upon cellular conditions that may be deemed too risky for continued DNA rearrangements.
Supported by NIA T32AG052363, NIH AI128137, OCAST
Sex-Dependent Differences in Serum Autoantibody Levels in the 3×Tg-AD Model of Alzheimer’s Disease
Authors: Ma Donglai , Song Wei , Michalski Bernadeta , Sakic Boris , Fahnestock Margaret 
Published: January 2023

Abstract
Sex-dependent discrepancies in disease prevalence and serum autoantibody levels are observed in patients and animal models of Alzheimer’s disease (AD). The present study examines whether gonadal hormones play a role in sex differences in serum autoantibody levels in the 3×Tg-AD mouse model of AD. 3×Tg-AD and wild-type (WT) mice were gonadectomised or sham-operated at 3 months of age. After behavioural phenotyping at 6 months of age, the animals were assessed for serum autoantibodies by indirect immunofluorescence for antinuclear antibodies (ANA) and by line-immunoblot assay for an additional 16 monospecific autoantibodies including anti-nucleosome antibodies. There were significant differences between the strains in ANA levels, with the major target antigens confirmed as nucleosomes. The results of ANA and anti-nucleosome assays were combined for further analysis. Further analysis revealed: 1) the level of serum autoantibodies in male 3×Tg-AD mice was higher than in female 3×Tg-AD animals, and this was not altered by orchiectomy. 2) sham-operated 3×Tg-AD female mice displayed a significantly lower level of serum autoantibodies than sham-operated WT females. 3) ovariectomy further reduced the level of serum autoantibodies in female 3×Tg-AD mice. The results suggest that dissimilar levels of serum autoantibodies in 3xTg-AD mice are a sex-dependent phenomenon and that female hormones play a role in regulation of their synthesis.
Funded by grant #SVB-158618 from the Canadian Institutes of Health Research to MF.
Interleukin-22 plays a protective role in animal model of experimental colitis by signaling on Math1+ cell.
Authors: Singh Ankita , Beaupre Michael , Lin Xun , Gaudino Stephen , Kumar Pawan 
Published: January 2023

Abstract
Interleukin-22 plays both beneficial and deleterious role in intestinal inflammation. Further investigation is required to understand cell target and mechanism of action of IL-22 in large intestine.
By using various intestinal cell specific IL-22Ra1 knockout mice models, such as entire intestinal epithelial cell (IL-22Ra1ΔIEC), Paneth (IL-22Ra1ΔDefa6), tamoxifen inducible Lgr5+ intestinal stem cell (IL-22Ra1ΔLgr5) and RU486 inducible secretory cell (IL-22Ra1ΔMath1-PGR). We tried to identify specific cell target of IL-22 in intestine. To induce experimental colitis, 8 days dextran sulphate sodium (DSS, 2%) was provided in drinking water followed by two days of normal water. On assessing colitis severity, more weight loss, histopathological changes and reduced colon length was observed in IL-22ra1ΔIEC mice. Increased expression of inflammatory cytokine genes and Lgr5+ stem cell related Olfm4 transcript were observed in distal colon of IL-22ra1ΔIEC as compared to IL-22ra1fl/fl mice. IL-22Ra1 signaling in IL-22Ra1ΔLgr5 and IL-22Ra1ΔDefa6 mice were dispensable in DSS induced colitis. Although goblet cell number was unaltered, expression of selective glycosyltransferases (causing O-glycosylation of mucin) were significantly reduced in colon of DSS treated IL-22Ra1ΔIEC as compared to IL-22Ra1fl/fl mice. Type-O glycan profile analysis of colon mucin using MALDI-TOF reveal premature termination of core 1 and 2 structure in IL-22Ra1ΔIEC mice as compared to IL-22Ra1fl/fl mice at steady state. Our preliminary data show that IL-22Ra1ΔMath1-PGR mice are more susceptible to DSS induced colitis.
IL-22 may act by inducing expression of glycosyltransferases in Math1+ cell to abrogate development of experimental colitis.
Arthroplasty surgery alters the local immune composition of the knee joint by causing increases in specific lymphocyte populations.
Authors: Cichos Kyle H , Raman Chander , Ghanem Elie S 
Published: January 2023

Abstract

Objective
We aimed to determine how the innate and adaptive immune cell composition and cytokine profiles of the knee joint microenvironment are altered in replaced knees compared to native knees.

Methods
Upon IRB approval, we obtained peripheral blood (PBMC) and synovial fluid (SF) from 14 native knees with osteoarthritis (OA) and 12 replaced knees at least 6 months after arthroplasty surgery, comprising 26 total patients. All PBMC and SF samples were analyzed by Flow Cytometry, and utilizing multidimensional reduction analysis via TriMap and clustering analysis by FlowSOM, cell clusters were phenotyped for identification of unique populations of cells. Additionally all samples were analyzed for cytokine/chemokine profiles using 65-Plex ProcartaPlex assays.

Results
Overall, there was no significant different in any of the innate cell populations studied, including neutrophils, monocytes (including G-MDSCs), macrophages (M1 of M2 phenotypes), and dendritic cells. B cells, CD8 effector T cells, CD4 T cells, Naive CD4 T cells, Th17 cells, and CD4 T cells with Th1-phenotypes s were all significantly increased in replaced knees compared to native knees (p &lt; 0.02 in each case). The replaced knee group showed elevated levels of CXCL5, HGF, IFN-gamma, IL-6, IL-16, CXCL10, CCL2 (MCP-1), CCL8, MDC, TNF-alpha, and VEGF-A compared to the native knee group (p &lt; 0.01 for all) and a decrease in CX3CL1, CXCL11, IL-3, IL-5, CXCL9, TRAIL (p &lt; 0.02 in all cases).

Conclusions
Arthroplasty surgery results in a lymphocytic shift in immune cell populations and a generally pro-inflammatory cytokine/chemokine profile. Overall, this lymphocytic shift may have implications for development of periprosthetic infections and warrants further study.
Interrogating mechanisms of CD8 T cell dysfunction in obese asthma
Authors: Hay Ceire , Sayed Samir , Conrey Peyton , Campos Jose , Pastore Christopher F , Sengupta Shaon , Herbert De’Broski , Henrickson Sarah E 
Published: January 2023

Abstract
Pediatric obese atopic asthma (OA) is a complex and poorly understood disease at the intersection of two of the most common chronic inflammatory diseases of childhood. Pediatric OA patients often experience severe asthma exacerbations caused by respiratory viral infections. However, the immunological mechanisms by which obesity modifies immune function in asthma remain poorly understood. Using high dimensional immune profiling, we have demonstrated CD8 T cells from pediatric OA patients have an exhausted-like immunophenotype compared to non-obese asthmatics. To more deeply characterize the role of CD8 T cell dysfunction in OA, we developed a mouse model of OA using high fat diet (HFD) to induce obesity and house dust mite (HDM) to induce asthma. We found that non-naïve CD8 T cells isolated from the lungs of OA mice demonstrate increased expression of multiple activation and inhibitory receptors (e.g., PD-1, CD39 and Tim3) as well as TOX, all consistent with T cell exhaustion, compared to healthy controls. We hypothesize the exhausted-like state of CD8 T cells in OA impairs anti-viral responses, thereby increasing severity of asthma exacerbation during respiratory viral infections. Ongoing work is testing antigen specific responses to influenza infection. Overall, this reveals a previously unrecognized role for exhausted-like CD8 T cell responses in mediating the immunopathogenesis of pediatric obese asthma.
This work was supported by the Translational Research Training Program in Environmental Health Sciences Pre-Doctoral Fellowship to C.A.H. and K08-AI135091 to S.E.H.
The NLRP3 Inflammasome Inhibitor OLT1177® Ameliorates Experimental Allergic Asthma in Mice
Authors: Lunding Lars P , Skouras Damaris B , Vock Christina , Dinarello Charles A , Wegmann Michael 
Published: January 2023

Abstract

Background
The symptoms of asthma arise on the basis of a chronic inflammation of the airways. The NLRP3 inflammasome plays a central role in regulating inflammation e.g. by activating interleukin (IL) 1 cytokines, which in turn promote allergic inflammation. This study aims to evaluate the therapeutic potential of the specific NLRP3 inhibitor OLT1177® on experimental allergic asthma (EAA) in mice.

Methods
The impact of OLT1177® on NLRP3 expression, caspase-1 activation, and IL-1β release were assessed in vivo and in allergen-restimulated mononuclear cells (MNC) from ovalbumin (OVA)-sensitized mice. The effect of intra-peritoneal OLT1177®-treatment on OVA- and house dust mite (HDM) EAA and on poly(I:C)-triggered exacerbation of EAA were assessed. Furthermore, the effect of oropharyngeal aspiration of OLT1177® and oral OLT1177® via an enriched diet were also evaluated in mice with OVA-induced EAA.

Results
OLT1177® reduced NLRP3 expression, caspase-1 activation, and levels of activated IL-1β in vitro and in mice with OVA-induced EAA. Intra-peritoneal treatment with OLT1177® significantly diminished allergic airway inflammation, mucus hyperproduction, and airway hyperresponsiveness (AHR) in mice with OVA- and HDM-induced EAA as well as in mice with poly(I:C)-triggered exacerbation of EAA. While oropharyngeal aspiration of OLT1177® had no effect on EAA, oral OLT1177® via feeding an enriched diet markedly reduced all pathophysiologic hallmarks of EAA in mice.

Conclusion
Intra-peritoneal and oral delivery of OLT1177® is effective to inhibit the NLRP3 inflammasome in vivo and to treat EAA in mice.
This study was mainly supported by the German Center for Lung Research (DZL). The study was also supported by Olatec Therapeutics LLC and the Interleukin Foundation.
GPR43 signaling in lung eosinophils suppresses neutrophilic airway inflammation in asthma
Authors: Yu Jihyun , Kim Seongryong , Kim You-Me 
Published: January 2023

Abstract
Asthma is caused by chronic inflammation in airways. It typically manifests type 2 immune activation and eosinophilia, while more severe, steroid-resistant asthma involves additional activation of Th17 cells and neutrophils. Even though pathogenic roles of eosinophils in asthma have been extensively studied, regulatory mechanisms curving down their overactivation are not well known. We found that eosinophils in asthmatic lung highly upregulate expression of GPR43, a receptor for short-chain fatty acid mediating various anti-inflammatory responses. In eosinophil-specific GPR43-deficient (GPR43ΔEOS) mice, asthma induction by Ova/alum or house dust mite extracts resulted in increased activation of Th17 cells and neutrophils in the lung compared to wild type mice. Consequently, GPR43ΔEOS mice showed more severe lung pathology and disease phenotype. Moreover, we identified a distinct Siglec-F+ neutrophil subset is enriched in GPR43ΔEOS mice by scRNA-seq analysis of the asthmatic lungs. Siglec-F+ neutrophils were recently shown to mediate Th17 cell activation and aggravation of asthma in vivo. We also found that Siglec-F+ neutrophils promote in vitro Th17 differentiation. Co-culture experiments of lung eosinophils and bone marrow neutrophils revealed that GPR43-deficient eosinophils directly mediate conversion of Siglec-F-neutrophils into Siglec-F+ neutrophils via GM-CSF secretion. Combined, our results demonstrate that GPR43 signaling in lung eosinophils prevents generation of Siglec-F+ neutrophils and Th17 cell activation, thereby suppressing severe asthma pathology.
Effect of HIV status on Th17/IL-17 immune function in subjects undergoing tuberculosis treatment in Kenya
Authors: Ongaya Asiko , Files Megan , Martinez-Martinez Yazmin Berenice , Chacha Robi , Kyalo Jeremiah , Mwangi Paul , Huante Matthew B , Navqi Kubra , Amukoye Evans , Endsley Janice J 
Published: January 2023

Abstract
Mycobacterium tuberculosis and HIV remain top global public health concern. Complexities associated with Mtb/HIV coinfections, including in those virally suppressed with anti-retroviral therapy (ART), present significant challenges to efforts to reduce both infections. The pattern of cytokines produced by T helper (Th) lymphocytes plays a central role in susceptibility, relapse and reinfection to TB. To understand how HIV and ART status affects recovery of Th cell effector function, we conducted a prospective study of HIV+ or HIV- subjects from Nairobi, Kenya. Blood samples were taken at start of TB treatment and 5 months after. Cytokine bead array was used to assess human Th cell-derived cytokines isolated from plasma from Nil, TB1 and Mitogen QuantiFERON tubes, including; IL-4, IL-9, IL-10, IL-17A, IL-17F, IL-22, IFN-γ and TNF-α. Intracellular cytokine staining (ICS) of cryopreserved peripheral blood mononuclear cells was also performed to determine changes in baseline numbers of Th1, Th2, Th17, Th22, IL-10+ Th cells, as well as HIV+ Th cells, across treatment groups. Results show reduced plasma IL-9, IFNg and IL-22 activation by mitogen in TB+HIV+ subjects that is independent of ART status, while IL-17A is moderately reduced in TB+HIV+ART- subjects. Cytokine recall to TB1 antigen was observed to be similar across all treatment groups, with the exception that a moderate reduction of IFNg and IL-17A was observed in the TB-HIV+ group. In subjects compliant for ART, ISC analysis revealed greater numbers of HIV+ cells in TB+HIV+, compared to TB-HIV+, groups. These preliminary results identify defects in effector cytokines produced by Th cells that may persist after ART and contribute to poorer TB treatment outcomes in subjects with HIV.
Supported by NIH, NIAID, 1R01AI147948
CD8+ T cell impairment drives severe human metapneumovirus respiratory disease in aged mice.
Authors: Parks Olivia Bartlett , Lan Jie , Zhang Yu , Sojati Jorna , Eddens Taylor John , Byersdorfer Craig A , Williams John V 
Published: January 2023

Abstract

Objective
Human metapneumovirus (HMPV) is a leading cause of severe respiratory disease in children &lt;5, adults &gt;65, and the immunocompromised. CD8+ T lymphocytes are the primary cell responsible for clearing HMPV infection in the lung. We tested the hypothesis that CD8+ T cell impairment in the aged host drives severe respiratory infection and delayed viral clearance.

Methods
B6 aged 72–73wk mice and young 6–7wk mice were infected with 2×106 PFU HMPV. Seven days post infection, mice were euthanized and lung, bronchoalveolar lavage (BAL), spleen, and draining lymph nodes were collected for flow cytometry. Lung and BAL supernatant was collected for viral titer, histology, and cytokine measurement.

Results
Aged mice lost more weight, had delayed viral clearance, and increased clinical scores and lung histopathology. Aged mice produced fewer HMPV-tetramer+ cells in lung and BAL. Aged CD8+ T lymphocytes in BAL co-expressed inhibitory receptors PD-1, TIM-3, LAG-3, and 2B4. Aged CD8+ T lymphocytes also upregulated senescence markers, T-bet and EOMES. Upon ex vivo peptide stimulation, aged CD8+ T cells produced significantly less granzyme B compared to young CD8+ T cells. Adoptive transfer of young T cells into aged mice rescued this severe disease phenotype. Young CD8+ cells in the aged host produced more tetramer+ cells and granzyme B and had less senescent marker expression.

Conclusion
Aged mice exhibit more severe respiratory disease when infected with HMPV. Aged CD8+ T cells appear to be impaired and upregulate senescence markers that contribute to severe HMPV infection in the aged host.
Supported by: NIH/NIAID 5R01 AI085062-09, John V. Williams, MD NIH T32GM008208 Medical Scientist Training Program, University of Pittsburgh School of Medicine
Helminth TGF-β mimic, TGM, increases leukocyte migration and activation while also enhancing cutaneous wound healing and tissue regeneration.
Authors: Lothstein Katherine E , Smyth Danielle J , Maizels Rick M , Gause William C 
Published: January 2023

Abstract
Intestinal parasites express excretory/secretory (ES) molecules, which modulate the type-2 immune response including anti-inflammatory and tissue repair mechanisms. TGF-β mimic (TGM), an ES molecule secreted by Heligmosomoides polygyrus (Hp), binds TGF-β receptors yet lacks structural homology to TGF-β and distinct receptor interactions. We now demonstrate that TGM enhanced wound healing using an in vivo wound biopsy model in which TGM, in a 1.5% carboxymethylcellulose solution, was topically administered beneath a Tegaderm layer. Through histological analysis, greater restoration of normal tissue structure in the wound beds of TGM treated mice was observed during mid to late stage wound healing. These observations included accelerated re-epithelialization as well as hair follicle regeneration, in the absence of increased scarring. Using flow cytometric, histological and gene expression analyses, differential expansion of myeloid populations at different stages of wound healing was observed. This included enhanced early accumulation and persistence of migratory Ly6C+ macrophages in TGM-treated wounds during the early inflammatory phase. Additionally, at mid-stage wound healing, we observed increased CD301b+ myeloid populations, a subset that is critical to efficient tissue restoration. In summary, TGM can enhance skin wound healing and associated recruitment of specific macrophage subsets and elevation of type 2 cytokines. This study indicates a role for TGM as a potential novel therapeutic option for enhanced wound healing.