bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dnase1L3 levels from macrophages in providing protection against SLE development
Authors: Hernandez Colton J , Engavale Minal B , Keyel Peter A 
Published: January 2023

Abstract
A hallmark symptom of SLE is elevated autoantibodies, especially anti-dsDNA antibodies in multiple organs and tissues. Deficiency of Dnase1L3, a serum endonuclease, allows the accumulation of self-DNA leading to autoantibody generation. The source of Dnase1L3 is primarily expressed by macrophages and dendritic cells. Complete Dnase1L3 deficiency is known to promote the SLE phenotype, however, the dosage required to prevent disease onset is unknown. We hypothesize that macrophages provide sufficient Dnase1L3 expression to regulate autoantibody levels in the host’s system. We generated conditional, macrophage specific Dnase1L3 knockout mice to assess disease progression. We collected serum samples from 6–50 weeks of age and measured total IgG and anti-dsDNA levels by ELISA to develop a longitudinal analysis of autoantibody elevation. At sacrifice, we observed pathology and measured IgG deposition in the kidney and heart by immunofluorescence. We observed that the elimination of Dnase1L3 in macrophages increased anti-dsDNA antibodies in the conditional knockout mice. In addition, we observed minor abnormalities in the glomeruli size for the conditional knockout mice as compared to the wild type mice. Our results indicate that macrophages are capable of providing protection against the development of SLE.
Human gingiva-derived mesenchymal stem cells ameliorate rheumatoid arthritis through directly targeting of rheumatoid arthritis synovial fibroblasts
Authors: Zheng Song Guo , Zhang Ximei , Jarjour Wael , Wang Julie 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is a systemic-autoimmune-mediated disease characterized by synovial hyperplasia and progressive destruction of joint. The aim of this study is to evaluate whether human gingiva derived MSCs (GMSCs) can alleviate rheumatoid arthritis through directly targeting of rheumatoid arthritis synovial fibroblasts (RA-SF) and underlying mechanism(s).
GMSCs or Primary Dermal Fibroblast (PDF) were co-cultured with LPS-stimulated RA-SF. The proliferation, migration and invasion abilities of RA-SF or GMSCs treated RA-SF were examined for determining the suppressive effects of GMSCs on RA-SF. To learn whether GMSCs suppress RA-SF migration and invasion in vivo, GMSCs were subcutaneously co-transplanted with RA-SF and health cartilage into SCID mice to develop a humanized synovitis model, H&E or immunofluorescence staining were employed to determine the effects. Moreover, GMSCs were transferred into collagen induced arthritis model (CIA), proliferation, migration and invasion capacities of synovial fibroblasts from model mice or GMSCs treated mice were compared.
We found that the proliferation, migration and invasion capacities of GMSCs treated RA SF were significantly declined. Importantly, the expression of IL-6, IL-8, MMP1, MMP2, MMP3 and MMP13 were dramatically reduced in GMSCs treated RA-SF (qPCR results). Administration of GMSCs was therapeutic in collagen induced arthritis synovial fibroblasts pathology (proliferation, migration and invasion). Furthermore, GMSCs directly suppress RA-SF invasion in a humanized synovitis model.
We conclude that the manipulation of GMSCs can ameliorate rheumatoid arthritis (RA) through directly targeting of rheumatoid arthritis synovial fibroblasts.
IRAK-M Down-regulates the Activation of Microglial NLRP3 Inflammasome and GSDMD-Mediated Pyroptosis in Microglial through Inhibiting IRAK1/TRAF6 Combination During Experimental Autoimmune Encephalomyelitis
Authors: Zhang Jun , Pei Shanshan , Ding Yuewen , Peng Yu , Chen Jinyu , Wang Haitao , Wang Honghao 
Published: January 2023

Abstract
The NOD-like receptor family pyrin domain-containing protein 3 (NLRP3) is a protein complex which mediates pro-inflammatory response and pyroptosis. The activation of NLRP3 inflammasome triggers pyroptosis via cleavage of the pore-forming protein gasdermin D. Previous studies have demonstrated that NLRP3 inflammasome activation in microglial contributes to the pathogenesis of EAE, while Inhibition of microglial NLRP3 inflammasome and the subsequent pyroptosis attenuates the severity of EAE. In addition, Interleukin-1 receptor-associated kinase (IRAK)-M, mainly expressed in microglial in the central nervous system, negatively regulates the TLR4/IL-1R signaling pathway through inhibiting the phosphorylation of IRAK1 and dissociation of IRAK1 from MyD88/IRAK-4/IRAK-1 complex as well as the formation of IRAK-1 and TRAF6 complex. Our recent study suggests the protective effect of microglial IRAK-M in EAE mice. Therefore, we hypothesize that IRAK-M may alleviate IRAK1-induced NLRP3 inflammasome activation and pyroptosis through inhibiting dissociation of IRAK1 from MyD88/IRAK-4/IRAK-1 complex and the following association of IRAK1 and TRAF6. Here, we used IRAK-M knockout mice and their microglia to clarify the role of IRAK-M in EAE. We demonstrated that IRAK-M decreased the incidence rate and improved the clinical symptoms accompanied by mild demyelination in EAE mice, and which was parallel to the inhibition of microglial NLRP3 inflammasome activation, pyroptosis, and IRAK1/TRAF6 formation in EAE. Our findings fill the gap in knowledge of IRAK-M on activation of NLRP3 inflammasome and pyroptosis and provide a potential novel target for the therapeutic strategy of multiple sclerosis and NLRP3-related diseases.
Retention and egress of Th1 cells in the inflamed skin
Authors: Salama Noah Alexander , Fowell Deborah J 
Published: January 2023

Abstract
The inappropriate trafficking and retention of immune subsets in tissue is a hallmark of many diseases prevalent in the developed world. These issues include autoimmunity, chronic inflammation, and unbalanced tissue repair responses which represent a significant disease burden: diabetes, coronary artery disease, inflammatory bowel disease, hepatitis, rheumatoid arthritis, and asthma. Although knowledge about cell trafficking to inflamed sites has grown in the past decade, there is a current gap in knowledge in identifying when and how immune cells decide to stay or leave these tissues.
To determine phenotypic and functional differences between those Th1 cells that remain at the inflamed site and those that exit, we employed the photoconvertible Kaede protein system to enable the labelling of Th1 cells in time and space. Kaede+OT-II Th1 T cells were adoptively transferred into WT mice immunized with CFA/OVA. Th1 cells recruited to the inflamed ear skin were photoconverted from Kaede green to Kaede red. Kaede red Th1 cells retained at the inflamed site could be distinguished from newly recruited Kaede green cells, while Kaede red Th1 cells that subsequently exited the inflamed skin were readily detected in the draining lymph node. Using this system, we are able to analyze the phenotypic (multicolor flow cytometry) and transcriptional signatures (RNA-seq) of Th1 cells that are retained at the inflamed site and those Th1 cells that exit (tissue-migrants). Given the temporal component of our model, we are gaining new insight into when and where expression of conventional markers of trafficking and activation are regulated (CD62L, CD44, and CD69) and seek to identify new pathways that dictate T cell retention and egress at inflammatory sites.
Epithelial cytokine IL-33 and sub-epithelial matrix protein tenascin C compartmentalize eosinophil phenotype and function
Authors: Nagasaka Reina , Doan Ton , Coden Mackenzie , Jeong Brian , Berdnikovs Sergejs 
Published: January 2023

Abstract
In allergic inflammation, eosinophils interacting with epithelial barriers experience exposure to epithelial cytokines and the extracellular milieu of airway remodeling. To better understand eosinophil activation biology in allergic tissue, we modeled the synergistic and opposing regulation of naïve eosinophils by both sub-epithelial extracellular matrix protein tenascin C (TNC), a key player in epithelial regeneration, and IL-33, an epithelial damage-induced cytokine. IL-5-differentiated bone marrow-derived murine eosinophils were cultured with recombinant TNC (25 ug/ml), IL-33 (100 ng/ml), or both for 24 hours and profiled by flow cytometry, qPCR, and DiffQuick staining. IL-33 alone significantly induced IL-4 and integrin CD11b expression, which was antagonized by co-culture with TNC. Conversely, TNC alone significantly promoted CD34 and intra-epithelial docking integrin CD103 expression, which was suppressed by IL-33 addition. Both treatments synergized in promoting expression of trans-epithelial migration marker CD11c. TNC promoted resident morphology while IL-33 induced an activated phenotype. This illustrates how microenvironmental cues may progressively shape eosinophils trafficking from sub-epithelial space across epithelial barriers.
Formin-like-1 mediates effector T cell extravasation through restrictive endothelial barriers enabling T cell entry into peripheral tissues and autoimmunity induction
Authors: Jacobelli Jordan , Thompson Scott B. , Sandor Adam M. , Lui Victor , Chung Jeffrey W. , Waldman Monique M. , Friedman Rachel S. 
Published: January 2023

Abstract
To perform their functions, T cells migrate from the blood vasculature into tissues by traversing the vascular wall through a process known as transendothelial migration (TEM). While many of the extracellular cues that guide this process have been identified, the role of downstream cytoskeletal effectors that mediate force generation and morphological changes required for TEM remains an area of active research. Formin-like-1 (FMNL1) is a cytoskeletal effector that mediates cytoskeletal remodeling. However, the role of FMNL1 in T cell functions, and migration in particular, is mostly unknown. The goal of our study was to examine the role of FMNL1 in T cell extravasation and trafficking. We generated a new FMNL1 knock-out (KO) mouse and identified a novel role for FMNL1 in promoting T cell migration through restrictive endothelial barriers during TEM. In vitro, FMNL1-deficient T cells had reduced actin polymerization in response to chemokine signaling. Furthermore, FMNL1 KO T cells had a selective impairment in chemotaxis through restrictive 3μm pores vs permissive 5μm pores. Using time-lapse microscopy, we determined that FMNL1-deficient T cells are severely impaired in their capacity to complete TEM through endothelial barriers. In vivo, we found that FMNL1 was dispensable for T cell extravasation through permissive high endothelial venules and homeostatic trafficking to lymphoid organs. Instead, FMNL1 regulated the ability of activated T cells to extravasate into inflamed peripheral tissues and enabled self-reactive T cells to induce autoimmune disease. Overall, our data identify FMNL1 as a key regulator of lymphocyte migration through restrictive barriers and as a potential target for modulating effector T cell trafficking.
Unraveling the role of β2 integrins in neutrophil migration to the lung and subsequent acute lung injury in sepsis
Authors: Koutsogiannaki Sophia , Kho Alvin T. , Yuki Koichi 
Published: January 2023

Abstract
Sepsis is the leading cause of death in intensive care units but not specific therapy is yet available. Acute lung injury (ALI) is a major complication in sepsis, resulting from massive migration of neutrophils to the lung, but the exact mechanism is not known. β2 (CD18) integrin family [consisting of αLβ2 (CD11a/CD18), αMβ2 (CD11b/CD18), αXβ2 (CD11c/CD18) and αDβ2 (CD11d/CD18)] and its counter-receptor on the endothelium, ICAM-1 (CD54), are critical for neutrophil migration, but their role in ALI is not yet delineated. Using a murine model of sepsis induced by cecal ligation and puncture (CLP) surgery, we showed decreased neutrophil levels and attenuated injury in the lung of β2−/− and ICAM-1−/−mice at 12h post-CLP, suggesting the importance of β2 integrins in this process. In addition, we observed decreased neutrophil levels and attenuated injury in the lung of αDβ2−/− mice but not in the lung of αLβ2−/− and αMβ2−/− mice, suggesting that among β2 integrins, αDβ2 has an orchestrating role in neutrophil migration to the lung and subsequent ALI in sepsis. In support, we found increased αDβ2 expression levels on neutrophils in the lung and blood of WT mice as sepsis progressed. RNAseq analysis in neutrophils from blood and lung of WT and αDβ2−/− mice showed that αDβ2 mediates neutrophil migration to the lung through pathways associated with antigen representation, maturation, apoptosis and NOD-like receptor signaling. αDβ2−/− neutrophils had also less Cxcr2, Ltb4r1 and Dhrs9 expression, associated with neutrophil migration and the retinoic acid pathway. Together, our results suggest a mechanism for neutrophil migration to the lung and subsequent ALI in sepsis in which αDβ2 has a major role and could be a novel target for therapeutic intervention.
BCG-Infected Dendritic Cells Induce TNF-alpha Dependent Cell Cluster Formation that Damage Brain Endothelial Cells in an In Vitro Model of the Blood Brain Barrier
Authors: Gilpin Trey E , Walter Fruzsina R , Herbath Melinda , Wigand Kelsey , Sandor Matyas , Fabry Zsuzsanna 
Published: January 2023

Abstract
Central nervous system tuberculosis (CNSTB) is the most serious manifestation of extrapulmonary tuberculosis infections representing 5–10% of all cases. How Mycobacterium tuberculosis (Mtb) crosses the highly regulated blood-brain barrier (BBB) and enters the CNS, is still not well understood. We showed previously that mycobacterium infected dendritic cells adhere to brain microvessel endothelial cells. Here, we applied a novel model of in vitro BBB consisting of primary brain astrocytes and microvessel endothelial cells to test the mechanisms of how mycobacterium infected dendritic cells (DCs) interact with peripheral blood mononuclear cells (PBMC) and affect the BBB. We show that infected DC migration was hindered but infected DCs induce cellular cluster formations containing PMBC and T cells in addition to DCs. These clusters contained 24% CD11c+, 57%CD11b+, and 16% CD4+ cells, while B220+ B cell number was negligible mimicking the cellular composition of pulmonary granulomatous-like lesions. PBMC and infected DC clusters strongly adhered to the in vitro BBB and induced cluster associated damage (CAD). CAD correlated with mitochondrial disorganization and the degradation of ZO-1, claudin-5 tight junction associated proteins, upregulation of cellular adhesion molecules VCAM-1 and ICAM-1 in endothelial cells. Anti-TNF-a inhibition reduced cluster formation as well as ICAM-1 expression on brain microvessel endothelial cells and mitigated CAD. This research will lead to better knowledge in understanding how mycobacteria affects the BBB that could lead to better therapeutic treatments for CNSTB infection.
Combining RNA in situ hybridization and spectral flow cytometry to investigate the leukocyte glycocalyx in autoimmunity
Authors: Marshall Payton L , Kaber Gernot , Linde Miles H , Barlow Graham L , Haddock Naomi L , Wagar Lisa , Nagy Nadine , Bollyky Paul L 
Published: January 2023

Abstract
The glycocalyx surrounding leukocytes has been studied for its role in immunomodulation and its potential as a therapeutic target. 4-methylumbelliferone (4MU) is a small molecule that inhibits production of the glycocalyx’s primary polysaccharide component hyaluronan (HA), by targeting the HA synthase (HAS). Oral treatment with 4MU has proved efficacious in preventing the development and progression of autoimmunity, though the mechanism is contested.
We have previously shown that 4MU strongly inhibits T-cell mediated autoimmunity, and report here that it also attenuates formation of antigen-specific plasmablasts and subsequent autoantibody production. This suggests that the HA glycocalyx plays a role in promoting the humoral component of autoimmunity.
The HAS proteins are deeply membrane embedded, preventing the use of traditional flow cytometry antibodies to define expression levels across leukocyte compartments. This limitation has made it difficult to identify cellular targets of 4MU. Alternative measurements must be used to study the HA glycocalyx. Surface HA content of a cell can be measured with labelled HA binding protein (HABP), though this does not explain whether HA was synthesized or scavenged with HA-binding receptors. RNA measurement can also be used to look at transcript levels of the HAS proteins.
Here, we combine RNA in situ hybridization, fluorescently labeled HA binding protein (HABP), and conventional flow cytometry antibodies to define the HA glycocalyx on leukocytes during autoimmunity. Utilizing Spectral Flow Cytometry, we can assess parameters in numbers comparable to Mass Cytometry (CyTOF) to identify which cells are most likely targeted by 4MU to prevent antibody formation during autoimmunity.
GPR56 inhibits chemotaxis of human Natural Killer and TEMRA lymphocytes
Authors: Janks Laura , Habiel David , Litvak Vladimir , Lin Lih-Ling , Kwon Hakju 
Published: January 2023

Abstract
Within the human immune system, the expression of GPR56, an adhesion G-protein couple receptor, is restricted to cytotoxic Natural Killer (NK) and T lymphocytes, including T effector memory re-expressing CD45RA (TEMRA) cells. In primary NK cells, GPR56 acts as an inhibitory receptor, as it diminishes their ability to eliminate target cells. Overexpression of GPR56 in immortal NK-92 cells also leads to robust inhibition of cell chemotaxis. GPR56 is speculated to promote retention of cytotoxic lymphocytes in inflamed peripheral tissues. Thus, GPR56 has the potential to regulate immune responses that may lead to excessive inflammation and autoimmunity. The aim of the present study was, therefore, to examine whether GPR56 mediates cytotoxic immune cell functions and whether GPR56 expression is altered in autoimmune diseases. To tackle these questions, we first compared expression of GPR56 in healthy and autoimmune disease patient tissues using scRNA-seq datasets. Next, we tested whether stimulation of GPR56 with a monoclonal antibody modulated the migratory capacity of NK and CD8+ TEMRA cells. We found that chemotaxis toward CXCL12, IL-8, and CX3CL1 was strikingly inhibited by GPR56. Using bulk RNA-seq, we found GPR56-stimulation impacted a multitude of signaling cascades that regulate cell chemotaxis. Taken together, our results confirm that GPR56 is a marker for NK and TEMRA cells in healthy and autoimmune patient tissues and establish GPR56 as a key regulator of lymphocyte chemotaxis.
The α7 nicotinic acetylcholine receptor is an important determinant of monocyte-derived macrophage migration
Authors: Pavlov Valentin A , Cui Kui , Addorisio Meghan , Williams David L , Yakubenko Valentin 
Published: January 2023

Abstract
Inflammation is regulated by the nervous system. The α7 nicotinic acetylcholine receptor (α7nAChR) on macrophages plays a key role in this regulation by meditating cholinergic suppression of pro-inflammatory cytokine release within the vagus nerve-based inflammatory reflex during endotoxemia and other conditions. However, the involvement of of α7nAChR in other macrophage functions remains poorly understood. The objective of this study was to provide insight into the role of α7nAChR in macrophage migration and accumulation in organs during murine endotoxemia. Briefly, monocyte progenitors isolated from bone marrow of wild type (WT) and α7nAChR knockout (KO) mice were labeled with red PKH26 (WT) or green PKH67 (α7nAChR−/−) fluorescent dyes, mixed in equal amounts and injected in tail vein of WT mice 20 min before LPS administration. 48 hours later, lungs, livers and spleens were isolated, digested and analyzed using multicolor flow cytometry (Fortessa-X20) and imaging flow cytometry (ImageStream X Mark II). α7nAChR deficiency resulted in significantly decreased accumulation of macrophages in liver (~10 folds), lung (~4 folds) and spleen (~3 folds) (P&lt;0.01). The morphology of migrated macrophage was verified by imaging flow cytometry. These results identify α7nAChR as an important determinant of the recruitment of monocytes to the site of acute inflammation. They also provide a rationale for further studies on the molecular mechanisms, including the involvement of adhesive receptors in α7nAChR-dependent monocyte/macrophage migration.
In vivo tracking: Using a photoconvertible system to probe immune cell entry and exit in the context of inflammation
Authors: Healey Alicia Marie , Fowell Deborah J 
Published: January 2023

Abstract
Chronic inflammatory diseases such as Crohn’s disease and rheumatoid arthritis affect millions of people worldwide. During an inflammatory response, immune cells enter the inflamed tissue to mediate pathogen clearance, however, their activation and overaccumulation in the tissue can lead to chronic inflammation and autoimmunity. Little is known about the signals that regulate immune cell entry into and exit from these inflamed tissues or about the retention cues that play a role in their accumulation. Therefore, a better understanding of these signals would inform the design of therapeutics intended to alleviate chronic inflammation. To study immune cell movement between a site of inflammation and lymphoid organs, our lab has acquired a mouse model that allows for the in vivo labeling of a subset of cells in time and space. The cells of Kaede transgenic mice constitutively express the photoconvertible fluorescent Kaede protein. CFA/OVA-inflamed ears were photoconverted from Kaede green to red by violet light exposure and the trafficking of Kaede red cells out of the skin to the draining lymph node was assessed over the course of 24 hours. Based on preliminary data, we found that, of the subsets assessed, neutrophils and CD4+ T cells made up the largest percentages of immune cells exiting the inflamed tissue on day 5 of inflammation. On the other hand, far fewer conventional dendritic cells and macrophages were seen to enter the draining lymph node from the inflamed tissue. The role of these tissue-emigrant immune cells is not yet clear, but cells such as neutrophils may play a role in shaping adaptive immune responses in the draining lymph node.
IL-11 induces surface marker expression and migration of T-lymphocytes
Authors: Seyedsadr Maryamsadat , Li Jian , Markovic-Plese Silva 
Published: January 2023

Abstract

Objective
To study the phenotypic and migratory changes of PBMCs following IL-11 stimulation

Background
Multiple sclerosis (MS) is presumably an autoimmune disease in which immune cells mediate neuroinflammation and neurodegeneration in central nervous system. Cytokines play a significant role in orchestrating the inflammatory response during the disease. Our lab has previously shown that in untreated MS patients, IL-11 is upregulated in the CSF and serum in comparison to matched healthy controls (HCs). Previous study has shown that CD4+IL11+ cells are accumulated in the CSF and the brain MS lesions, suggesting that they play a role in the lesion formation.

Methods
14 untreated RRMS patients and age-, sex- and race-matched HCs were enrolled in the study upon signing an Institutional Review Board consent forms.

Results
By using flow cytometry we characterized IL-11-secreting CD4+ lymphocytes and show that they have increased expression of CCR6, CCR4, ICAM-1, IL-11RA, GM-CSF, and IFNγ in comparison to CD4+IL-11− cells (p values&lt;0.03). In vitro studies further showed that stimulation of PBMCs with recombinant human IL-11, upregulated the expression of CCR4 and CCR6 (p values&lt;0.03, 1.2 and 1.4 fold changes) in CD4+ gated cells. In a trans-well system we observed that IL-11 can act as a chemoattractant for anti-CD3/CD28 mAb stimulated PBMCs and increase their migration (p value=0.04, migration index=1.2) in both patients and HCs.

Conclusion
We observed that IL-11 induces migratory phenotype in CD4+ lymphocytes in patients with RRMS, which can be explored as a therapeutic target in early RRMS.