CCL2 Levels Research Articles

P0198 Comparative Transcriptomic Analysis of Tonsil-derived and Adipose-derived Mesenchymal Stem Cells: Proliferation, Migration, and Anti-inflammatory Potential in Murine Colitis Model

Abstract Background Tonsil-derived mesenchymal stem cells (TMSCs) have emerged as a promising alternative to traditional MSCs due to their rapid proliferation and accessibility. This study explores the unique transcriptomic and cellular properties of TMSCs compared with adipose-derived MSCs (AMSCs). Methods TMSCs and AMSCs were cultured under identical conditions in low-glucose DMEM with 10% FBS. Proliferation was evaluated via CCK-8 assays, and migration potential was assessed using scratch assays. Transcriptomic differences were analyzed using RNA sequencing, with key findings validated by qPCR. Results TMSCs displayed distinct morphological features compared to AMSCs, with TMSCs showing a spindle shape, while AMSCs appeared more irregular and rounded shape. In terms of proliferation, TMSCs demonstrated a significantly higher proliferation rate by day 7 (245.3% of control vs. 134.4% of control, p < 0.001). However, there was no significant difference in migration capacity between TMSCs and AMSCs over a 24-hour period (50.8 ± 8.2 vs. 53.8 ± 9.8, p = 0.537). RNA sequencing analysis revealed notable differences in gene expression, with TMSCs showing enrichment in pathways related to extracellular matrix, cell adhesion, and immune response, particularly with upregulation of the MAPK signaling pathway compared with AMSCs. For anti-inflammatory cytokines, IL-33, FOXF1, and HGF were upregulated in TMSCs relative to AMSCs. In relation to proliferation and migration, CCL2, CXCL12, and STC1 levels were also higher in TMSCs. Additionally, TMSCs exhibited increased expression of RAB27B, suggesting a greater potential for exosome release. In a DSS-induced acute colitis model, both TMSCs and AMSCs alleviated colitis symptoms with significant improvements in colon length (p < 0.05). Real-time PCR analysis of mouse colonic tissue also demonstrated a reduction in pro-inflammatory cytokines, such as IL-1β, with TMSCs showing a more pronounced reduction. Conclusion This study demonstrates that TMSCs possess unique characteristics, including a higher proliferation rate and distinct gene expression profiles compared to AMSCs. While both MSC types exhibited similar anti-inflammatory effects in murine colitis model, the enhanced anti-inflammatory potential and increased exosome release capacity of TMSCs, combined with their easy availability, highlight their promise for clinical application.

P0130 Assessment of macrophage types in the intestinal tissue of treatment naïve Inflammatory Bowel Disease patients in relation to their disease and activation of the CCL2/CCR2 pathway

Abstract Background Resident intestinal macrophages play a key role in maintaining gut homeostasis and exhibit a hypoactive tolerogenic phenotype. Recently, several subsets of intestinal macrophages have been identified based on CD11c expression which differ in their inflammatory and functional profiles. Notably, pro-inflammatory CD11c+ monocyte-like macrophages are found elevated in patients with inflammatory bowel disease (IBD), specifically Ulcerative colitis (UC) and Crohn’s disease (CD). These cells are recruited from blood monocytes via the CCR2/CCL2 chemokine pathway. Our study is focused on observing the patterns of CCL2-driven blood monocyte migration and their translation into subsets of intestinal tissue macrophages that can determine the degree of gut inflammation in newly diagnosed patients. Methods Patients referred with suspected IBD were seen in an inception clinic. Intestinal biopsies and matched blood samples were obtained from pre-treatment UC and CD patients during their index colonoscopy. Those with inflammation excluded during this procedure formed a ‘non-IBD’ control group. Biopsy segments were cultured to generate conditioned media and also digested for flow cytometry analysis. Plasma concentrations of CCL2 and intestinal secretome were measured using ELISA or Olink proteomic analysis respectively. Efficacy of blood monocyte chemotaxis towards CCL2 was assessed in the presence or absence of CCL2/CCR2 antagonists and the profile of monocyte subsets in whole blood were determined using whole blood flow cytometry analysis. Results As previously described, we observed distinct CD11c+ and CD11c- macrophage populations in IBD intestinal tissue, with expansion of the CD11c+ subset within inflamed regions which expressed higher levels of the chemokine receptors CCR2 and CX3CR1. Higher concentrations of CCL2 were observed in conditioned media from inflamed IBD tissue compared to non-inflamed regions and this correlated with elevated numbers of circulating blood monocytes. Chemotaxis assays revealed a corresponding increase in CCL2-induced monocyte migration in patients with IBD compared to the ‘non-IBD’ control group. Conclusion Our study provides further insights into the role of the CCL2/CCR2 chemokine axis in driving monocyte recruitment and their differentiation into pro-inflammatory intestinal macrophages in IBD. We demonstrate that inflamed intestinal regions exhibit elevated levels of CCL2 correlating with an expansion of the CD11c⁺ macrophage subset and increased CCL2-driven monocyte migration in these subjects. These findings underscore the significance of targeting the CCL2/CCR2 pathway and that it might be an important driver in the development IBD.

ARTN and CCL23 predicted chemosensitivity in acute myeloid leukemia: an Olink® proteomics approach

BackgroundThe standard “7 + 3” induction results in 30% of de novo acute myeloid leukemia (AML) patients not achieving complete remission (CR). We aimed to utilize the Olink® platform to compare the bone marrow plasma proteomic profiles of newly diagnosed de novo AML patients who did and did not achieve CR following “7 + 3” induction treatment.MethodsThis prospective study included 43 untreated AML patients, stratified into CR (n = 29) and non-CR (n = 14) groups based on their response to “7 + 3” induction therapy. We employed the Olink® Explore-384 Inflammation platform for proteomic analysis to investigate differences in bone marrow plasma protein levels between the CR and non-CR groups.ResultsProteomic analysis demonstrated that the CR group exhibited significantly higher bone marrow plasma levels of ARTN and CCL23 than did the non-CR group. Immunohistochemical staining confirmed a higher proportion of tissue samples with intense staining for ARTN (25.40% vs. 7.05%, p = 0.013) and CCL23 (24.14% vs. 14.29%, p = 0.039) in the CR group. These findings were corroborated by bulk-RNA-seq, which indicated significantly elevated mRNA expression levels of ARTN (1.93 vs. -0.09; p = 0.003) and CCL23 (1.50 vs. 0.12; p = 0.021) in the CR group. The Human Protein Atlas provided external support for our findings.ConclusionsThe results suggest that ARTN and CCL23 may serve as biomarkers for predicting responsiveness to the “7 + 3” induction in untreated AML. Using an enzyme-linked immunosorbent assay to identify the roles of ARTN and CCL23 in predicting AML chemosensitivity may enhance clinical applicability in the future.

IL-25 Enhances B Cell Responses in Type 2 Inflammation Through IL-17RB Receptor.

Alarmin cytokine IL-25 promotes type 2 inflammatory responses in disorders such as asthma and chronic rhinosinusitis with nasal polyps (CRSwNP) and known targets include ILC2 and Th2 cells. However, other cellular targets for IL-25 remain poorly defined. To investigate induction and expression of IL-25 receptor (IL-17RB) by B cells and evaluate responsiveness of IL-17RB-expressing B cells to IL-25 invitro. IL-17RB expression, regulation and function on B cells were evaluated in peripheral blood-derived B cells by flow cytometry and RT-PCR, including in response to IgE-inducing stimuli (anti-CD40 mAb and IL-4). Single-cell RNA sequencing was used to compare IL-17RB+ and IL-17RB-activated peripheral blood-derived B cells. To evaluate B cell IL-17RB expression within type 2 inflamed tissue, B cells were compared from nasal polyps, control turbinate tissue and matched peripheral blood. Activation of B cells with anti-CD40 and IL-4 increased IL-17RB expression at both protein and mRNA level, which was further upregulated by IL-25. B cells induced to express IL-17RB responded to IL-25 with enhanced antibody production. Single-cell RNA-sequencing showed that IL17RB+ activated B cells expressed higher levels of IGHE, CCL17 and CCL22 compared to IL17RB- B cells. B cells from nasal polyp tissue expressed higher levels of surface IL-17RB compared with control tissue, correlating with patient-reported CRSwNP severity (SNOT-22). Peripheral blood B cells activated under IgE-inducing conditions express surface IL-17RB, and tissue IL-17RB+ B cells are increased in type 2 inflammation. IL-17RB+ cells have a distinct transcriptional profile and respond to IL-25 with enhanced antibody production, highlighting the IL-25/IL-17RB pathway as a potential therapeutic target for CRSwNP and other type 2 inflammatory disorders.

Downregulation of CCR2 reduces ventricular remodeling after myocardial infarction by splenic nerve neuromodulation in acute and chronic rat models.

Pathological remodeling after myocardial infarction (MI) confers the development of heart failure. Our prior research has indicated that splenic nerve neuromodulation mitigates myocardial ischemia-reperfusion injury (IRI) by reducing levels of proinflammatory factors. This study aims to explore the potential therapeutic benefits of splenic nerve neuromodulation in MI and the underlying mechanism. Splenic nerve neuromodulation was performed through electrical splenic nerve stimulation (SpNS). In the acute myocardial IRI model, post-mortem analyses encompassed RNA sequencing and a range of molecular biology techniques, with the application of CCR2 antagonists (RS-504393) to inhibit the CCR2. In the chronic MI model, rats underwent echocardiographic assessment four weeks post-MI, after which tissues were harvested. In the acute IRI model, the negative regulation of chemokines production pathway was enriched by RNA-seq, and SpNS reduced the levels of CCR2, CCL2, and CCL7. The administration of RS-504393 decreased cardiomyocyte apoptosis, reduced myocardial damage, and lowered proinflammatory cytokines levels following myocardial IRI. Additionally, SpNS was shown to inhibit oxidative stress, proinflammatory cytokine levels, and cardiac collagen deposition, as observed four weeks post-MI. SpNS also restrained sympathetic nerve remodeling and improved left ventricular function, in part by downregulating CCR2 in the chronic MI model. SpNS demonstrated significant improvements in cardiac function, reductions of cardiac remodeling and inhibitions of excessive sympathetic activation in the chronic MI model by downregulation of CCR2. Our study provides novel evidence that splenic nerve neuromodulation may serve as a potential therapeutic intervention in MI patients.

Diagnostic Markers of Severe COVID-19 and Community-Acquired Pneumonia in Children From Southern India.

COVID-19 severely impacts children in India, with many developing severe pneumonia or multisystem inflammatory syndrome (MIS-C). Concurrently, non-COVID-19 respiratory viruses causing community-acquired pneumonia (CAP) have resurged. These conditions present similarly, challenging accurate diagnosis. This study aims to compare inflammatory markers and clinical parameters in children with severe COVID-19 pneumonia, non-COVID-19 CAP, and COVID-associated MIS-C. We assessed 12 mediators in serum from 14 children with severe COVID-19 pneumonia, 16 with severe non-COVID-19 CAP, and 9 with MIS-C. Clinical characteristics and routine laboratory findings at admission were recorded. Children with MIS-C had significantly higher levels of IL-1RA, IL-8, and TNF compared with those with severe COVID-19 pneumonia; and higher levels of CCL2, HGF, M-CSF, and IL-8 compared with severe non-COVID-19 CAP. GROα levels tended to be higher in severe COVID-19 pneumonia. Clinical presentations were similar, but MIS-C patients had distinct laboratory findings, including lower platelet counts and albumin levels, and higher creatinine and liver enzyme levels. MIS-C exhibited a unique inflammatory profile. IL-8 emerged as a potential biomarker for MIS-C, while increased GROα levels in severe COVID-19 pneumonia merit further exploration. Combining inflammatory markers with routine laboratory parameters may improve the diagnosis and differentiation of these conditions, enhancing patient management.

Identification of Immune Infiltration-Associated CC Motif Chemokine Ligands as Biomarkers and Targets for Colorectal Cancer Prevention and Immunotherapy.

Colorectal cancer (CRC) is the third most common cancer globally, with limited effective biomarkers and sensitive therapeutic targets. An increasing number of studies have highlighted the critical role of tumor microenvironment (TME) imbalances, particularly immune escape due to impaired chemokine-mediated trafficking, in tumorigenesis and progression. Notably, CC chemokines (CCLs) have been shown to either promote or inhibit angiogenesis, metastasis, and immune responses in tumors, thereby influencing cancer development and patient outcomes. However, the diagnostic and prognostic significance of CCLs in CRC remains unclear. In this study, multiple online tools for bioinformatics analyses were utilized. The findings revealed that the mRNA expression levels of CCL3, CCL4, and CCL26 were significantly elevated in CRC tissues compared to normal tissues, whereas CCL2, CCL5, CCL11, CCL21, and CCL28 mRNA levels were markedly downregulated. Additionally, dysregulation of CCL4, CCL5, and CCL21 was strongly associated with clinical staging, and elevated levels of CCL4, CCL11, and CCL28 were linked to significantly prolonged survival in CRC patients. Functional enrichment analysis indicated that the cellular roles of CCLs were predominantly associated with the chemokine, Wnt, and Toll-like receptor signaling pathways, as well as protein kinase activity. Furthermore, transcriptional regulation of most CCLs involved RELA and NFKB1. Key downstream targets included members of the SRC family of tyrosine kinases (HCK, LYN, and LCK), serine/threonine kinases (ATR and ATM), and others such as CSNK1G2, NEK2, and CDK2. Moreover, CCLs (CCL2, CCL3, CCL4, CCL5, CCL11, CCL21, and CCL28) exhibited strong correlations with major infiltration-related immune cells, including B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils, and dendritic cells. In conclusion, our study provides novel insights into the potential utility of CCLs as biomarkers and therapeutic targets for CRC prevention and immunotherapy.

Constructing the Well Regenerated Decellularized Adipose Tissue Using External Volume Expansion Device.

External volume expansion (EVE) devices has been demonstrated to enhance the survival of fat grafts. Decellularized adipose tissue (DAT) serves as a promising scaffold for adipose regeneration; however, the effectiveness of adipose regeneration in DAT remains limited, and the underlying mechanisms of its regeneration require further investigation. This study explores the potential of EVE technology to enhance DAT-mediated adipogenesis by facilitating cellular recruitment and establishing a microenvironment conducive to adipose tissue regeneration. DAT was injected into the dorsal area of rats, followed by daily treatment with an EVE suction device for 10 hours per day over 14 days. Control groups underwent transplantation without suction. After the treatment period, tissue samples were collected and analyzed. This included volume measurement, H&E staining, immunofluorescence staining for CD34, CD90, CD68, CD31, and perilipin, electron microscopy for microscopic analysis, and ELISA analysis for IL1, TNFα, CCL2, and CXCL12. Fourteen days post-transplantation, the volume of DAT significantly increased in the EVE group compared to the control group. Histological H&E staining revealed a higher peripheral region in the EVE group. Electron microscopy examination showed that EVE suction led to increased porosity in the DAT material, with a greater number of cells adhering to the material. Immunofluorescence staining for CD34/CD90 adipose-derived stem cells also showed a significant increase in the EVE group. The presence of CD68-positive macrophages increased after EVE suction. Evaluation of vascularization using CD31 staining showed a higher level of vascularization in the EVE group compared to the control group. ELISA analysis of IL-1, TNF-α, CCL2, and CXCL12 levels demonstrated that the EVE group effectively increased the levels of adipogenic factors within the DAT. EVE enhances DAT-mediated adipogenesis by promoting stem cell recruitment, macrophage activation, and adipogenesis-related cytokine expression, ultimately improving the regeneration of functional adipose tissue. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

CCL24 and Fibrosis: A Narrative Review of Existing Evidence and Mechanisms.

Tissue fibrosis results from a dysregulated and chronic wound healing response accompanied by chronic inflammation and angiogenesis. Regardless of the affected organ, fibrosis shares the following common hallmarks: the recruitment of immune cells, fibroblast activation/proliferation, and excessive extracellular matrix deposition. Chemokines play a pivotal role in initiating and advancing these fibrotic processes. CCL24 (eotaxin-2) is a chemokine secreted by immune cells and epithelial cells, which promotes the trafficking of immune cells and the activation of profibrotic cells through CCR3 receptor binding. Higher levels of CCL24 and CCR3 were found in the tissue and sera of patients with fibro-inflammatory diseases, including primary sclerosing cholangitis (PSC), systemic sclerosis (SSc), and metabolic dysfunction-associated steatohepatitis (MASH). This review delves into the intricate role of CCL24 in fibrotic diseases, highlighting its impact on fibrotic, immune, and vascular pathways. We focus on the preclinical and clinical evidence supporting the therapeutic potential of blocking CCL24 in diseases that involve excessive inflammation and fibrosis.

5β-hydroxycostic acid from Laggera alata ameliorates sepsis-associated acute kidney injury through its anti-inflammatory and anti-ferroptosis effects via NF-κB and MAPK pathways.

The whole plant of Laggera alata is frequently utilize to remedy inflammatory diseases including nephritis as a traditional Chinese medicine. However, its active ingredients and mechanism of action against sepsis-associated acute kidney injury (SA-AKI) are unknown. This study aimed to identify active compounds from L. alata that inhibit renal inflammation and ameliorate SA-AKI, and to elucidate their mechanisms of action. The chemical constituents were separated from the ethyl acetate layer of L. alata methanol extract by column chromatography over silica gel, medium-pressure liquid chromatography and semipreparative high-performance liquid chromatography. Extensive spectroscopic techniques were applied to determine the chemical structures. The anti-inflammatory efficiency was measured by analyzing the NO production in RAW 264.7 cells. The levels of IL-6, IL-1β, CCL-2 and CCL-5 mRNA were determined by qRT-PCR. Cecal ligation and puncture (CLP) surgery is a frequently applied method to establish the mouse sepsis model. Sepsis was thus induced in mice via CLP. The effect in the treatment of SA-AKI was evaluated by H&E staining and ELISA detection. Western blotting was used to evaluate the protein levels involved in ferroptosis, NF-κB and MAPK signaling pathways. Twelve compounds were obtained from L.alata including four unreported sesquiterpenoids (1-4). Compound 5 exhibited the most significant inhibitory effect on NO production with the IC50 value of 6.034 μM and could restrain the mRNA expression of inflammatory factors IL-6, IL-1β, CCL-2 and CCL-5. The in vivo results demonstrated that compound 5 alleviated the renal injury by decreasing the serum IL-6, IL-1β, Cr, and BUN levels, reducing the kidney contents of Cys-C and KIM-1, and regulating the kidney levels of MDA, GSH, ferrous iron, GPX4, FTH1, and SLC7A11. Furthermore, Compound 5 also repressed the NF-κB and MAPK pathways in vitro and in vivo. This study revealed that compound 5 could ameliorate SA-AKI through exerting its anti-inflammatory and anti-ferroptosis effects via NF-κB and MAPK pathways. The current research supported the traditional use of L.alata in the treatment of renal diseases.

Trabectedin enhances the antitumor effects of IL-12 in triple-negative breast cancer.

Interleukin-12 (IL-12) is a potent NK cell-stimulating cytokine, but the presence of immunosuppressive myeloid cells such as myeloid-derived suppressor cells (MDSC) can inhibit IL 12-induced NK-cell cytotoxicity. Thus, we hypothesized that trabectedin, a myeloid cell-depleting agent, would improve the efficacy of IL-12 in triple-negative breast cancer (TNBC). In vitro treatment of healthy donor NK cells with trabectedin increased expression of the activation marker CD69 and mRNA expression of T BET (Tbx21), the cytotoxic ligands TRAIL (TNFSF10) and Fas ligand (FASLG) and the dendritic cell (DC)-recruiting chemokine lymphotactin (XCL1). The combination of IL-12 and trabectedin increased NK-cell cytotoxicity, activation and production of IFN-γ, TNF-α and granzyme B in the presence of human TNBC cells. Treatment of 4T1 and EMT6 tumor-bearing mice with IL-12 and trabectedin led to a significant reduction in tumor burden compared to single-agent controls, and the highest levels of plasma IFN-γ, intratumoral CD8+ T cells and conventional type 1 DC. MDSC and M2-like macrophages were significantly decreased with combination therapy. NK-cell depletion abrogated the effects of combination therapy, as did elimination of CD8+ T cells. NK-cell depletion led to lower levels of the NK cell-derived chemokine CCL5 and the DC-derived chemokine CXCL10, higher tumor burden, and decreased intratumoral CD8+ T cells. IL 12 and trabectedin also significantly enhanced the response of TNBC to anti-PD-L1 therapy. These data suggest that MDSC depletion augments the ability of IL-12-activated NK cells to drive the infiltration of DC and CD8+ T cells into TNBC for an antitumor effect.

Inflammatory changes in the choroid plexus following subarachnoid hemorrhage: the role of innate immune receptors and inflammatory molecules.

The choroid plexus is located in the cerebral ventricles. It consists of a stromal core and a single layer of cuboidal epithelial cells that forms the blood-cerebrospinal barrier. The main function of the choroid plexus is to produce cerebrospinal fluid. Subarachnoid hemorrhage due to aneurysm rupture is a devastating type of hemorrhagic stroke. Following subarachnoid hemorrhage, blood and the blood degradation products that disperse into the cerebrospinal fluid come in direct contact with choroid plexus epithelial cells. The aim of the current study was to elucidate the pathophysiological cascades responsible for the inflammatory reaction that is seen in the choroid plexus following subarachnoid hemorrhage. Subarachnoid hemorrhage was induced in rats by injecting non-heparinized autologous blood to the cisterna magna. Increased intracranial pressure following subarachnoid hemorrhage was modeled by using artificial cerebrospinal fluid instead of blood. Subarachnoid hemorrhage and artificial cerebrospinal fluid animals were left to survive for 1, 3, 7 and 14 days. Immunohistochemical staining of TLR4, TLR9, FPR2, CCL2, TNFα, IL-1β, CCR2 and CX3CR1 was performed on the cryostat sections of choroid plexus tissue. The level of TLR4, TLR9, FPR2, CCL2, TNFα, IL-1β was detected by measuring immunofluorescence intensity in randomly selected epithelial cells. The number of CCR2 and CX3CR1 positive cells per choroid plexus area was manually counted. Immunohistochemical changes were confirmed by Western blot analyses. Immunohistochemical methods and Western blot showed increased levels of TLR9 and a slight increase in TLR4 and FRP2 following both subarachnoid hemorrhage as well as the application of artificial cerebrospinal fluid over time, although the individual periods were different. The levels of TNFα and IL-1β increased, while CCL2 level decreased slightly. Accumulation of macrophages positive for CCR2 and CX3CR1 was found in all periods after subarachnoid hemorrhage as well as after the application of artificial cerebrospinal fluid. Our results suggest that the inflammation develops in the choroid plexus and blood-cerebrospinal fluid barrier in response to blood components as well as acutely increased intracranial pressure following subarachnoid hemorrhage. These pro-inflammatory changes include accumulation in the choroid plexus of pro-inflammatory cytokines, innate immune receptors, and monocyte-derived macrophages.

Cytokine Profile in Children Following SARS-CoV-2 Infection: Preliminary Findings.

We provide preliminary evidence that, also in children, Long coronavirus disease (COVID) may be characterized by a proinflammatory signature. Ten Long COVID patients, 7 convalescent subjects after COVID infection and 4 healthy controls were enrolled. When adjusted for sex, children with long COVID had statistically significant differences in the levels of Flt3L, CD5, uPA, CCL23, CD40 and TGFα. When adjusted for age, CCL23 levels remained statistically significant.

Single-nucleus transcriptome profiling provides insights into the pathophysiology of adhesive arachnoiditis.

Adhesive arachnoiditis (AA) is a rare form of chronic degenerative pathology associated with persistent inflammation in the arachnoid matter of the spinal cord. Despite the existing knowledge, the detailed pathological mechanisms underlying AA are not fully understood. This study aimed to elucidate through comprehensive single nuclei RNA sequencing (snRNA-seq) to delineate the transcriptomic landscape of AA. From six arachnoid membrane samples, a total of 52,886 cells met the quality control standards for analysis. The main cell populations identified with specific gene markers were as follows: fibroblasts, glial cells, microglial cells, endothelial cells, mural cells, plasma cells, and T cells. Downstream analysis of fibroblasts, glial cells, and microglial cells was performed. Notably, fibroblast subsets 1 and 3 demonstrated a strong association with AA. Among them, subcluster 3 demonstrated elevated expression of genes COL1A1, COL3A1, and FN1, indicative of enhanced Wnt/β-catenin and extracellular matrix (ECM) synthesis pathways. Subcluster 3 was predicted to progressively transform into subcluster 1. In subcluster 1, there was a significant upregulation of genes such as BMP and ALPL, signaling enhanced activation of calcification-related pathways. This was highly relevant to end-stage arachnoid ossification formation. After being activated, microglial cells transformed into inflammatory disease-associated microglial cells and continued to express high levels of chemokines CCL2, CCL4, IL-1β, and other inflammatory factors NAMPT, INPP5D and NLRP3. This might be the main reason why AA recurrence is frequently observed in patients. These insights enhance our understanding of the pathological progression of AA and may contribute to the identification of novel therapeutic targets.

Differentiating multiple sclerosis with predominant spinal cord and optic nerve involvement from other autoimmune demyelinating diseases using B cell immunophenotyping and gene expression profiling.

Multiple sclerosis (MS) may present with predominant involvement of the spinal cord and optic nerve (MS/w-SCON) and mimic other autoimmune inflammatory demyelinating disorders (AIDD) such as neuromyelitis optica spectrum disorder (NMOSD), and relapsing inflammatory optic neuritis (RION). Thus, biomarkers are required for effective differential diagnosis of AIDD. Patients with MS/w-SCON (n = 20), MS without involvement of SCON (MS/wo-SCON) (n = 22), NMOSD (n = 16), RION (n = 15) and healthy individuals (n = 21) were included. Peripheral blood cell immunophenotypes were analyzed by flow cytometry and gene expression levels of isolated B cells were assessed by microarray analysis and quantitative real-time PCR. Significantly increased Breg, plasmablast, and plasma cell ratios distinguished MS/w-SCON from other groups. Most differentially expressed B cell genes were subjected to protein-protein interaction yielding a network of 13 immune mediators (IL18, IL18R1, P2RY12, CSF3R, TNFSF4, TNFRSF13C, TNFRSF1A, CCL20, CCL5, CCR4, CCL4L2, CXCL5, CXCL10). CCL4L2 and CCR4 expression levels distinguished MS/w-SCON. IL18/IL18R1 and CCL5 were distinguishing factors for NMOSD and RION, respectively. Peripheral blood B cell expression levels of CCL4L2, CCR4, IL18, IL18R1 and CCL5 are candidate biomarkers for differential diagnosis of AIDD of CNS. Our results substantiate the significance of B cells in the pathogenesis of these disorders.

Innate immune response after BNT162b2 COVID-19 vaccination associates with reactogenicity

BackgroundThe innate immune response is important for the development of the specific adaptive immunity, however it may also be associated with reactogenicity after vaccination. We explore the association between innate responsiveness, reactogenicity, and antibody response after first COVID-19 vaccination. MethodsWe included 146 healthy Dutch individuals aged 12–59 who received their first BNT162b2 (Comirnaty, Pfizer) COVID-19 vaccination. Data on reactogenicity were collected for each individual through daily questionnaires from day 0–5 after vaccination. From 60 participants, serum (adults) and plasma (adolescents) samples were collected before and/or 2 ± 1 days after vaccination to measure cytokines/chemokines as markers for innate responsiveness. Each individual was categorised into innate low, intermediate and high responder based on above or below the median value for each analyte detected after vaccination. For 137 participants, serum was collected at day 28 after vaccination for Spike S1- and RBD-antibody concentration. The associations between reactogenicity and/or innate responsiveness and/or log-transformed antibody concentration were explored using logistic and linear regressions. ResultsMost participants (85 %) reported both local and systemic symptoms after vaccination. Two participants reported no symptoms. More than half (54 %) reported one or more moderate symptoms. Significantly higher levels of pro-inflammatory mediators CXCL9, CXCL10, CXCL11, IFNγ and CCL20 in adults, and CXCL9, CXCL10 and CXCL11 in adolescents, were found after vaccination. Participants who showed high innate immune responsiveness had higher odds (OR 6.0; 95 % CI 1.4–33) of experiencing one or more moderate symptoms. No association was found between innate responsiveness or having one or more moderate symptoms with Spike S1- or RBD-antibody concentration at day 28 after vaccination. ConclusionOur results suggest an association between the strength of the innate immune response and the severity of reactogenicity to SARS-CoV-2 vaccination. However, more research is needed to understand the relation between reactogenicity and immunogenicity of COVID-19 vaccines.

The Role of CCL18 in Rheumatoid Arthritis Diseases

Rheumatoid Arthritis (RA) is a chronic auto-immune disease that mostly occurs in joint space with remarkably painful swelling. RA detail pathologies remain unclear, but several C-C motif chemokines factors and Interleukin have been identified as associated factors and correlated disease-associate activities (such as CCL2, CCL18, IL-6). Among these factors, increasing evidence indicates CCL18 could be an appropriate drug target, as it is highly correlated with disease-associated activities (DAS28) (elevated level of CCL18 in both patient serum and local synovial fluids). High-concentration CCL18 in synovial fluids has been reported to facilitate pro-inflammation factor production, finally resulting in painful swelling in joint space. In vivo studies showed significant drug efficiency with anti-TNF-a treatment (CCl18 suppression). Most current approved drugs target TNF, IL-6, or other factors, but do not target CCL18 directly or indirectly. From this perspective, CCL18 has been believed to be a new target for RA therapeutic drug development.

Regulation and Function of CCL2 and N-Myc in Retinoic Acid-treated Neuroblastoma Cells.

Treatment with retinoic acid (RA) often promotes neuroblastoma differentiation and growth inhibition, including the suppression of the expression of the MYCN oncogene. However, RA also targets protumoral chemokines, such as CCL2, which may contribute to the development of resistance. The present study aimed to investigate the regulation and function of CCL2 and N-Myc in RA-treated neuroblastoma cells. In Kelly or SH-SY5Y cells, viability was quantified by cell fitness assays. Expression was analyzed using quantitative PCR and the regulation of proteins using enzyme-linked immunoabsorbent assays (ELISA) or western blots. In MYCN-amplified Kelly cells, endogenous CCL2 levels were significantly lower compared to MYCN non-amplified SH-SY5Y cells. Treatment with 5 μM RA increased CCL2 release in both cell lines, but reduced N-Myc levels and cell numbers in Kelly cells. Over-expression of MYCN enhanced viability in SH-SY5Y cells, but did not affect RA-induced CCL2 release, while supplementation of CCL2 in Kelly cells did not prevent RA-mediated growth reduction. Impaired N-Myc or CCL2 signaling reduced the survival of all RA-treated cells and inhibition of N-Myc also decreased CCL2 levels. However, attenuated survival signaling was not generally associated with reduced levels of N-Myc or CCL2. Co-application of RA and the growth factor receptor inhibitors cediranib or crizotinib decreased N-Myc levels only in Kelly cells, while CCL2 release was dependent on the cell type and stimulus. CCL2 and N-Myc promote the viability of RA-treated cells, although the levels of these mediators were not consistently correlated with cellular outcomes, especially during apoptotic signaling.

Prognostic Value of CXCL13, CCL11, and CCL20 Chemokines in Multiple Sclerosis.

Objective: The course of relapsing-remitting multiple sclerosis (RRMS) is highly variable and there is a lack of effective prognostic biomarkers. This study aimed to assess the potential prognostic value of the chemokines B lymphocyte chemoattractant molecule (CXCL13), eotaxin-1 (CCL11), and macrophage inflammatory protein 3-alpha (CCL20) in RRMS. Methods: Forty-two patients with MS were enrolled, along with 22 controls, 12 of the controls were idiopathic intracranial hypertension (IIH) patients, and 10 of the controls were other neurologic diseases (OND). Chemokine levels were measured using enzyme-linked immunosorbent assay (ELISA) in serum and cerebrospinal fluid (CSF) samples. Results: No significant differences were observed among the groups in serum levels of CXCL13, CCL11, and CCL20 (p = 0.509, p = 0.979, p = 0.169, respectively). CSF CXCL13 levels were significantly higher in the OND group (p = 0.016). A PATH analysis showed CSF CXCL13 was significantly associated with new T2 hyperintense lesions on brain magnetic resonance imaging (p < 0.001), and baseline serum CCL11 levels were associated with EDSS (p = 0.030), implying its potential role in indicating neurodegenerative processes and possible progression risk. Serum CCL20 correlated with EDSS (p = 0.002) and lesion burden (p < 0.001), reflecting disease severity. Conclusions: These findings suggest that CSF CXCL13 could serve as a useful biomarker for predicting active disease in RRMS, while follow-up serum CCL11 may assist in identifying progression. Although these chemokines are not specific to MS, higher levels may signal disease activity, severity, and transition to more progressive stages.

Prolyl hydroxylase domain inhibitors prevent kidney crystal formation by suppressing inflammation.

The early stages of kidney crystal formation involve inflammation and hypoxia-induced cell injury; however, the role of the hypoxic response in kidney crystal formation remains unclear. This study investigated the effects of a prolyl hydroxylase domain inhibitor (roxadustat) on renal calcium oxalate (CaOx) crystal formation through in vitro and in vivo approaches. In the in vitro experiment, murine renal tubular cells (RTCs) were exposed to varying roxadustat concentrations and CaOx crystals. CaOx monohydrate (COM) crystal adhesion was evaluated using fluorescent labels, whereas western blotting was used to examine protein expression. Quantitative real-time polymerase chain reaction was used to analyze gene expression changes. Macrophage responses were investigated by co-culturing them with RTCs treated with COM. In the in vivo experiment, C57BL/6J mice were injected with roxadustat or saline for 2days, followed by glyoxylate for 6days to induce renal crystal deposition. Biochemical measurements recorded plasma erythropoietin, urinary data, and pH levels. Roxadustat suppressed the adhesion of COM crystals to RTCs and the expression of proinflammatory genes, such as chemokine (C-C motif) ligand 2 (Ccl2) and secreted phosphoprotein 1 (Spp1). Roxadustat decreased the expression levels of Ccl2, tumor necrosis factor (Tnf), and interleukin6 (Il6) in co-cultured macrophages. In the in vivo experiment, the amount of renal CaOx crystal deposits was significantly lower in the roxadustat-treated group than in the vehicle group. Roxadustat treatment decreased Ccl2, Tnf, and adheision G protein-coupled receptor E1 (Adgre1) expression in the kidneys. Roxadustat reduced kidney inflammation and CaOx crystal deposition, suggesting its potential as a therapeutic option for kidney stone prevention.