CXCL10 Levels Research Articles

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 .

CXCL13 levels in cerebrospinal fluid in relapsing-remitting multiple sclerosis: The role of Borrelia in neuroinfections.

This study was compared the Borrelia antibodies and chemokine ligand 13 (CXCL13) levels in cerebrospinal fluid (CSF) samples from cases diagnosed with relapsing-remitting multiple sclerosis (RRMS), radiologically isolated syndrome (RIS), and pseudotumour cerebri (PTC). A total of 43 CSF samples were collected from patients diagnosed with RRMS, RIS and PTC. We prospectively investigated Borrelia IgG and IgM antibodies in the CSF samples of the cases by enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) method, and CXCL13 levels by ELISA. Data were statistically analysed using the the Spearman rank correlation test. Five antigens (protein 19, 20, 21, 58, and outer surface protein C (OspC)) were positive due to confirmation of the positive samples for Borrelia antibodies by the WB method. There were no significant differences in CSF CXCL13 levels between the three groups. The CXCL13 level was found to be statistically higher in the demyelinating group compared to the PTC group (p=0.001). The Borrelia antibodies were found positive in CSF samples of RRMS patients. The coexistence of high CXCL13 (may be a potential biomarker) suggests that LNB may also play a role in the etiopathogenesis of RRMS. In addition, the positive detection of OspC and p58 WB bands in most cases suggests that these protein bands can be used as in the differential diagnosis of neurodegenerative diseases and Lyme disease.

Imbalanced VWF-ADAMTS13 axis contributes to the detrimental impact of a preceding respiratory tract infection on stroke.

Respiratory tract infections (RTIs) caused by bacteria or viruses are associated with stroke severity. Recent studies have revealed an imbalance in the von Willebrand factor (VWF)-ADAMTS13 axis in patients with RTIs, including COVID-19. We examined whether this imbalance contributes to RTI-mediated stroke severity. Wild-type (WT), Vwf -/-, or Adamts13-/- mice with respective littermate controls (Vwf +/+, or Adamts13+/+) were infected intranasally with sublethal doses of S. aureus (on days 0, 2, and 5) or mouse-adapted SARS-CoV-2 (on day 0) and subjected to transient (30 or 45 min) cerebral ischemia followed by reperfusion. In S. aureus-infected mice, infarct volumes were assessed on day 2 and functional outcomes on weeks 1 and 4 post-reperfusion. In SARS-CoV-2-infected mice, infarct volumes and functional outcomes (Bederson score) were assessed on day 1 post-reperfusion. We demonstrated that S. aureus or SARS-CoV-2 RTI was accompanied by an imbalance in the VWF-ADAMTS13 axis and an increase in plasma levels of IL-6, CXCL1, and MCP-1, which was associated with larger infarcts and worse functional outcomes (P<0.05 vs. mock-infection). S. aureus- or SARS-CoV-2-infected Vwf -/- mice exhibited reduced infarcts and improved functional outcomes, while infected Adamts13-/- mice displayed greater stroke severity (P<0.05 vs. control). In the models of RTI preceding stroke, VWF contributes to stroke severity, while ADAMTS13 is protective.

CXCL12 ameliorates neutrophilia and disease severity in SARS-CoV-2 infection.

Neutrophils, particularly low-density neutrophils (LDNs), are believed to contribute to acute COVID-19 severity. Here, we showed that neutrophilia can be detected acutely and even months after SARS-CoV-2 infection in patients and mice, while neutrophil depletion reduced disease severity in mice. A key factor in neutrophilia and severe disease in infected mice was traced to the chemokine CXCL12 secreted by bone marrow cells and unexpectedly, endothelial cells. CXCL12 levels were negatively correlated with LDN numbers in longitudinal analyses of patient blood samples. CXCL12 blockade in SARS-CoV-2-infected mice increased blood/lung neutrophil numbers thereby accelerating disease progression without changing lung virus titers. The exaggerated mortality caused by CXCL12 blockade can be reversed by neutrophil depletion. In addition, blocking interactions between SARS-CoV-2 and Angiotensin-Converting Enzyme 2 (ACE2) reduced CXCL12 levels, suggesting a signal transduction from virus-mediated ACE2 ligation to increased CXCL12 secretion. Collectively, these results demonstrate a previously unappreciated role of CXCL12 in diminishing neutrophilia, including low density neutrophilia, and its deleterious effects in SARS-CoV-2 infections. The results also support the involvement of SARS-CoV-2-endothelial cell interactions in viral pathogenesis.

CXCL5 as a biomarker for early diagnosis and prognosis of sepsis: A comprehensive clinical evaluation.

Sepsis, a critical condition caused by a dysregulated host response to infection, has high morbidity and mortality rates. Timely diagnosis and treatment are vital for improving patient outcomes. This study explores the potential role of CXCL5 in the diagnosis, severity assessment, and prognosis of sepsis. We included 147 sepsis patients, 50 patients with systemic inflammatory response syndrome (SIRS) and 120 healthy controls. Serum CXCL5 levels, inflammation scores (APACHE II, SOFA), and other laboratory indicators were recorded. Univariate and multivariate logistic regression analyses were conducted to assess the relationship between CXCL5 and sepsis diagnosis, severity, and prognosis. A prognostic nomogram was constructed and evaluated using receiver operator characteristic curves, calibration curves, and clinical decision curves. Serum CXCL5 levels in sepsis patients were significantly higher than those in patients with SIRS and healthy controls. CXCL5 was identified as a risk factor for sepsis diagnosis. CXCL5 levels were significantly elevated in patients with septic shock (P=0.04) and in deceased patients compared to survivors (P<0.001). The prognostic model, incorporating CXCL5, lactate, APACHE II scores, C-reactive protein levels, and respiratory rate, demonstrated high predictive accuracy with an area under the curve of 0.873. Calibration and decision curve analyses demonstrated the model's good predictive performance and potential clinical value. Serum CXCL5 concentration is a promising biomarker for enhancing the diagnostic accuracy and prognostic evaluation of sepsis. The constructed multivariate prediction model offers new insights into sepsis prognosis, but its direct application in clinical practice requires further validation.

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.

Investigating the Role of Quercetin, an Active Ingredient in Bazhen Decoction, in Targeting CXCL8 to Inhibit Macrophage M2 Polarization and Reshape the Immunological Microenvironment of Colorectal Cancer.

Bazhen Decoction (Eight Treasures Decoction) has demonstrated efficacy in the treatment of colorectal cancer (CRC), yet the active ingredients in it and the mechanisms underlying their anti-cancer properties are not well understood. Through network pharmacology, the effective components of Bazhen Decoction against CRC and their corresponding key genes were delineated. Molecular docking was executed to identify the active component targeting the key gene CXCL8, which led to the discovery of Quercetin. The cellular thermal shift assay method was then used to verify the binding interaction. CRC cells were treated with incremental concentrations of Quercetin, cell viability was evaluated by the Cell Counting Kit-8 assay to calculate the IC50, and apoptosis rates were determined by flow cytometry. Expression of the apoptosis-related proteins Bcl-2 and Cleaved caspase-3 was measured using western blot. The impact of Quercetin on macrophage polarization was studied by co-culturing the treated CRC cells with macrophages, assessing M1 and M2 macrophage distribution via flow cytometry, and quantifying cytokine levels (IL-6, IL-10, IL-12, and CXCL8) with enzyme-linked immunosorbent assay (ELISA). The active ingredient Quercetin from Bazhen Decoction exhibited a targeted binding affinity with the key gene CXCL8, which enabled it to inhibit the proliferation of CRC cells and induce cell apoptosis. The overexpression of CXCL8 was associated with the promotion of CRC malignancy, yet the presence of Quercetin could lessen the impact of CXCL8 overexpression on CRC cells. Moreover, the treatment with Quercetin leads to a diminished abundance of M2 macrophages and an increase in the levels of cytokines IL-6 and IL-12, while reducing the levels of IL-10 and CXCL8, which indicates that Quercetin has an inhibitory effect on macrophage M2 polarization. Quercetin, the active component in Bazhen Decoction that is known for anti-CRC effects, targets and inhibits CXCL8 to impede the malignant behaviors and the M2 polarization of macrophages. Thus, Quercetin may be utilized as an immunomodulatory agent in CRC treatment.

DOC2b enrichment mitigates proinflammatory cytokine-induced CXCL10 expression by attenuating IKKβ and STAT-1 signaling in human islets.

Type 1 diabetic human islet β-cells are deficient in double C 2 like domain beta (DOC2b) protein. Further, DOC2b protects against cytokine-induced pancreatic islet β-cell stress and apoptosis. However, the mechanisms underpinning the protective effects of DOC2b remain unknown. Biochemical studies, qPCR, proteomics, and immuno-confocal microscopy were conducted to determine the underlying protective mechanisms of DOC2b in β-cells. DOC2b-enriched or -depleted primary islets (human and mouse) and β-cell lines challenged with or without proinflammatory cytokines, global DOC2b heterozygous knockout mice subjected to multiple-low-dose-streptozotocin (MLD-STZ), were used for these studies. A significant elevation of stress-induced CXCL10 mRNA was observed in DOC2b-depleted β-cells and primary mouse islets. Further, DOC2b enrichment markedly attenuated cytokine-induced CXCL10 levels in primary non-diabetic human islets and β-cells. DOC2b enrichment also reduced total-NF-κB p65 protein levels in human islets challenged with T1D mimicking proinflammatory cytokines. IKKβ, NF-κB p65, and STAT-1 are capable of associating with DOC2b in cytokine-challenged β-cells. DOC2b enrichment in cytokine-stressed human islets and β-cells corresponded with a significant reduction in activated and total IKKβ protein levels. Total IκBβ protein was increased in DOC2b-enriched human islets subjected to acute cytokine challenge. Cytokine-induced activated and total STAT-1 protein and mRNA levels were markedly reduced in DOC2b-enriched human islets. Intriguingly, DOC2b also prevents ER-stress-IKKβ and STAT-1 crosstalk in the rat INS1-832/13 β-cell line. The mechanisms underpinning the protective effects of DOC2b involve attenuation of IKKβ-NF-κB p65 and STAT-1 signaling, and reduced CXCL10 expression.

PDGFRα-positive cell-derived TIMP-1 modulates adaptive immune responses to influenza A viral infection.

Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a physiologic inhibitor of the matrix metalloproteinases (MMPs), but little is known about the role of TIMP-1 in regulating the pathogenesis of influenza A virus (IAV) infection. Here, we performed both in vivo and in vitro experiments to investigate the regulation and function of TIMP-1 during IAV infection. Specifically, plasma levels of TIMP-1 are significantly increased in human subjects and wild-type (WT) mice infected with 2009 H1N1 IAV compared with levels in uninfected controls. Also, TIMP-1 is strikingly upregulated in PDGFRα positive (PDGFRα+) cells in IAV-infected murine lungs as demonstrated using conditional KO (cKO) mice with a specific deletion of Timp-1 in PDGFRα+ cells. Our in vitro data indicated that TIMP-1 is induced by transforming growth factor-β (TGF-β) during lipofibroblasts (lipoFBs)-to-myofibroblast (myoFB) transdifferentiation. Timp-1 deficiency protects mice from H1N1 IAV-induced weight loss, mortality, and lung injury. IAV-infected Timp-1-deficient mice showed increased macrophages, and B and T cell counts in bronchoalveolar lavage (BAL) on day 7 postinfection (p.i.), but reduced BAL neutrophil counts. Increased Cxcl12 levels were detected in both BAL cells and lungs from Timp-1-deficient mice on day 3 p.i. Taken together, our data strongly link TIMP-1 to IAV pathogenesis. We identified that PDGFRα-lineage cells are the main cellular source of elevated TIMP-1 during IAV infection. Loss of Timp-1 attenuates IAV-induced mortality and promotes T and B cell recruitment. Thus, TIMP-1 may be a novel therapeutic target for IAV infection.NEW & NOTEWORTHY Our data strongly link tissue inhibitor of metalloproteinases-1 (TIMP-1) to influenza A virus (IAV) pathogenesis. TIMP-1 is highly increased in PDGFRα-lineage cells during IAV infection. Transforming growth factor-β (TGF-β) induces TIMP-1 during lipofibroblast (lipoFB)-to- myofibroblast (myoFB) transdifferentiation. Timp-1 deficiency protects mice from H1N1 IAV-induced weight loss, mortality, and lung injury. TIMP-1 may be a novel therapeutic target for IAV infection.

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 &lt; 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 &lt; 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.

Distinct pathophysiological pathways support stratification of Sjögren's disease based on symptoms, clinical, and routine biological data.

Recently, three distinct phenotypes of Sjögren's disease (SjD) patients have been described, based on cluster analysis: B-cell active with low symptoms (BALS), high systemic activity (HSA), and low systemic activity with high symptoms (LSAHS). We aimed to assess whether these clusters were associated with distinct biomarkers and the prognostic value of IFN signature. The ASSESS cohort is a 20-year prospective cohort of SjD patients. The following biomarkers were compared: IFN-α2, IFN-γ, CXCL10, CXCL13, BAFF, IL7, FLT3, CCL19, and TNFRII. IFN signature was assessed using transcriptomic analysis. We then compared systemic and symptomatic evolution, and the risk of new immunosuppressant prescription and of lymphoma, according to the IFN signature across the three clusters. 395 patients (94% female, median age 53 [43-63] years) were included. Higher levels of CXCL-13, IL7, and TNF-RII levels were found in the BALS and HSA clusters compared to the LSAHS cluster. A high IFN signature was mainly found in the BALS cluster (57%, vs. 48%, and 38% in the HSA and LSAHS clusters, respectively). This IFN signature was mainly driven by type I IFN, with higher levels of IFN- α2. In the BALS cluster, a high IFN signature was associated with a higher risk of new immunosuppressant treatment (HR 9.38; 95% CI 1.22-72.16). All lymphoma occurred in patients with high IFN signature. The three SjD clusters displayed distinct expression of IFN signature, and markers of T- and B-cell activation, confirming distinct pathophysiological mechanisms. High IFN signature could predict systemic evolution in the BALS cluster.

Evaluation of the effectiveness of combined therapy with intravenous immunoglobulin and plasmapheresis in patients with steroid-resistant form of pemphigus based on the cytokine and chemokine profiles assessment

BACKGROUND: Pemphigus is a serious life-threatening disease characterized by the formation of IgG autoantibodies against the cell membranes, leading to the formation of intraepidermal blisters. AIM: To evaluate the effectiveness of combined therapy with intravenous immunoglobulin and plasma exchange for steroid-resistant patients with pemphigus based on the cytokine, chemokine and granulysin profiles investigation. MATERIALS AND METHODS: The group of patients receiving systemic glucocorticoid monotherapy (Group 1; control group) consisted of 26 patients with pemphigus vulgaris. The group of steroid-resistant patients (Group 2; main group) who received combined therapy with systemic glucocorticoid, intravenous immunoglobulin (IVIg), and plasmapheresis included 15 people. The presence of steroid resistance was assessed by Murrell consensus (2008). All pemphigus patients received the initial dose of systemic glucocorticoids of 80–100 mg/day with subsequent slow tapering, according to guidelines. The treatment protocol for combined therapy included four sessions of discrete plasma exchange per week every other day. Immediately after completion of the plasma exchange cycle, IVIg was added to the ongoing treatment, with a total dose of 2 g/kg per cycle. The levels of IL-4, IL-10, IL-15, TNF-α, chemokines CXCL8, CCL11, and granulysin were assessed via ELISA method. RESULTS: We observed some discrepancies in cytokine profiles in both groups of patients. In patients who received combined therapy, there was a statistically significant decrease in the levels of IL-4, IL-15, TNF-α compared to those in patients on systemic glucocorticoid monotherapy ― IL-4, IL-15 TNF-α (p 0.01). Notably, that the level of CCL11 in serum of steroid-resistant patients before the IVIg therapy was significantly higher (Me=51 pg/ml) compared to systemic glucocorticoid monotherapy group (Me=10 pg/ml; p 0.01). The level of granulysin after the treatment with IVIg and plasma exchange in group 2 was also significantly lower (Me=0 ng/ml) compared to the group of control (Me=2700 ng/ml respectively; p 0.01). CONCLUSION: We found a trend towards higher serum levels of IL-4, IL-15, and CCL11 in steroid-resistant pemphigus patients who received combined therapy with IVIg and plasma exchange compared to the control group. Moreover, these cytokines can be considered as the potential biomarkers for refractory disease course, and might be used as therapeutic targets in the future. It should be also noted that the prolonged remission of patients receiving combined therapy with systemic glucocorticoid, IVIg, and plasma exchange, was on average two years.

Inflammaging Markers in the Extremely Cold Climate: A Case Study of Yakutian Population.

Yakutia is one of the coldest permanently inhabited regions in the world, characterized by a subarctic climate with average January temperatures near -40 °C and the minimum below -60 °C. Recently, we demonstrated accelerated epigenetic aging of the Yakutian population in comparison to their Central Russian counterparts, residing in a considerably milder climate. In this paper, we analyzed these cohorts from the inflammaging perspective and addressed two hypotheses: a mismatch in the immunological profiles and accelerated inflammatory aging in Yakuts. We found that the levels of 17 cytokines displayed statistically significant differences in the mean values between the groups (with minimal p-value = 2.06 × 10-19), and 6 of them are among 10 SImAge markers. We demonstrated that five out of these six markers (PDGFB, CD40LG, VEGFA, PDGFA, and CXCL10) had higher mean levels in the Yakutian cohort, and therefore, due to their positive chronological age correlation, might indicate a trend toward accelerated inflammatory aging. At the same time, a statistically significant biological age acceleration difference between the two cohorts according to the inflammatory SImAge clock was not detected because they had similar levels of CXCL9, CCL22, and IL6, the top contributing biomarkers to SImAge. We introduced an explainable deep neural network to separate individual inflammatory profiles between the two groups, resulting in over 95% accuracy. The obtained results allow for hypothesizing the specificity of cytokine and chemokine profiles among people living in extremely cold climates, possibly reflecting the effects of long-term human (dis)adaptation to cold conditions related to inflammaging and the risk of developing a number of pathologies.

Treatment Response Biomarkers for Systemic Sclerosis-Associated Interstitial Lung Disease.

This studied investigated whether changes in circulating biomarkers predict progressive pulmonary fibrosis (PFF) in patients with systemic sclerosis-associated interstitial lung disease (ILD) receiving treatment. Participants of Scleroderma Lung Study (SLS) II, which compared mycophenolate (MMF) versus cyclophosphamide (CYC) for SSc-ILD, who had blood samples at baseline and 12-months were included. Levels for C-reactive protein (CRP), interleukin (IL)-6, chemokine ligand 4 (CXCL4), chemokine ligand 18 (CCL18)and Krebs von den Lungen 6 (KL-6) were measured, and a logistic regression model evaluated relationships between changes in these biomarkers and the development of PPF by 24 months. Ninety-two of the 142 randomized participants had longitudinal biomarker measurements and the required clinical outcome data, with 19 (21%) meeting criteria for PPF. In the whole cohort, changes in KL-6 levels were significantly correlated with PPF. KL-6 increased in patients who developed PPF and decreased in patients who did not (Mean change 365.68 [SD 434.41]) vs -207.45 [SD 670.26]; P<0.001). In the MMF alone arm, changes in CRP and CXCL4 were also significantly correlated with PPF. When added to an existing prediction model based on baseline factors associated with PPF in this cohort (sex, baseline reflux severity and CXCL4 levels), the change in KL-6 remained significantly associated with PFF (OR 1.4; P=0.0002). Changes in the circulating levels of KL-6 after treatment with MMF or CYC predicted PPF, even after adjusting for baseline factors associated with PPF. Measuring longitudinal KL-6 in patients with SSc-ILD may improve how we personalize therapy in SSc-ILD.

Repigmentation in non-segmental vitiligo using the Janus kinase inhibitor upadacitinib, a retrospective case series.

Vitiligo is the most frequently diagnosed depigmentation disease, affecting nearly 0.5-2% of individuals worldwide. This disorder is characterized by melanocyte loss, which results in skin and hair depigmentation. Psychological problems are common in patients, especially in those with the involvement of the face and hands. Several studies have recently focused on the use of JAK inhibitors for vitiligo treatment. However, studies on the selective JAK1 inhibitor upadacitinib for vitiligo treatment are limited. This study aimed to assess the efficacy and safety of upadacitinib in the treatment of vitiligo. This retrospective case series included five patients diagnosed with non-segmental vitiligo who were treated with upadacitinib for 4 months or longer. Disease severity was assessed using the Vitiligo Area Scoring Index (VASI). Five patients took 15mg of upadacitinib orally for 4 consecutive months, and all achieved repigmentation. The plasma collected from the peripheral blood of the four patients showed that the CXCL9 level dropped after upadacitinib treatment. However, the CXCL10 level did not change significantly using enzyme-linked immunosorbent assay (ELISA). Flow cytometry revealed that the ratio of CD4 + CD3+/CD8 + CD3 + T cells in the blood samples tended to decrease through the treatment. And the Th1-like Tregs (CD4 + Foxp3 + IFN-γ + Tregs) were also downregulated in the peripheral blood. In conclusion, the JAK1 inhibitor upadacitinib was found to be effective and safe for treating non-segmental vitiligo.

Involvement of CXCL12/CXCR4 in CB2 receptor agonist-attenuated morphine tolerance in Walker 256 tumor-bearing rats with cancer pain

While low-dose cannabinoid 2 (CB2) receptor agonists attenuate morphine tolerance in cancer pain models, chemokine ligand 12 (CXCL12)/chemokine receptor 4 (CXCR4) expression induces morphine tolerance. Whether CB2 receptor agonists attenuate morphine tolerance by modulating CXCL12/CXCR4 signaling or whether CXCL12/CXCR4 signaling affects the mu opioid receptor (MOR) in the development of morphine tolerance in cancer pain remains unclear. In this study, we investigated the attenuation of morphine tolerance by a non-analgesic dose of the CB2 receptor agonist AM1241, focusing specifically on the modulation of CXCL12/CXCR4 signaling and its effect on the MOR. Rats received intrathecal Walker 256 tumor cell implantations and were treated with morphine combined with the intrathecal injection of AM1241 or the CB2 receptor antagonists AM630 and AM1241, or a CXCL12-neutralizing antibody, exogenous CXCL12, or the CXCR4 antagonist AMD3100. Our results show that CXCL12 and CXCR4 levels increased significantly in morphine-tolerant rats and were reduced by AM1241 pretreatment, which was reversed by AM630. CXCL12/CXCR4 expression accelerated the development of morphine tolerance and downregulated MOR expression. CXCR4 colocalized with MOR and CB2. Therefore, a non-analgesic dose of AM1241 attenuated morphine tolerance via CXCL12/CXCR4 signaling, whereas CXCL12/CXCR4 signaling participated in the development of morphine tolerance, potentially by modulating MOR expression in Walker 256 tumor-bearing rats.Graphical

CD69+CD103+CD8+ tissue-resident memory T cells possess stronger anti-tumor activity and predict better prognosis in colorectal cancer

BackgroundColorectal cancer (CRC) is one of the most prevalent cancers worldwide. Despite advancements in therapeutic methodologies, it still causes a high rate of patient mortality. CD8+ tissue-resident memory T (TRM) cells are strategically positioned to mediate effective anti-tumor responses. However, the characteristic surface molecules and functions of CD8+ TRM cells exhibit significant heterogeneity.MethodsThe roles and anti-tumor biological functions of different CD8+ TRM subsets in CRC were determined by clinical CRC samples, bioinformatics analysis, and in vitro experiments including co-culture experiments and transwell migration assays. The signaling pathways that synergistically regulate the differentiation of CD8+ TRM cells were identified by in vitro CD8+ T cell activation and inhibition assays, and the functioning transcription factors were predicted using the UCSC and JASPAR databases.ResultsWe found that different CD8+ TRM subsets existed in CRC tumor tissues, which were identified as CD69−CD103−CD8+ TRM, CD69+CD103−CD8+ TRM (SP CD8+ TRM), and CD69+CD103+CD8+ TRM (DP CD8+ TRM) subsets. Compared with SP CD8+ TRM cells, increased infiltration of DP CD8+ TRM cells predicted better prognosis and played a protective role mainly in tumor invasion and lymph node metastasis of CRC. DP CD8+ TRM cells expressed higher levels of effector molecules and exerted stronger anti-tumor effects in a FAS/FASL pathway-dependent manner. Additionally, DP CD8+ TRM cells secreted higher levels of CXCL13 and recruited B cells into tumor tissues through the CXCL13/CXCR5 signaling axis to form tertiary lymphoid structures, participating in anti-tumor immune responses. Notch and TGF-β signaling pathways synergistically regulate the differentiation of DP CD8+ TRM cells.ConclusionsWe clarified the roles and mechanisms of different CD8+ TRM subsets in CRC and identified that DP CD8+ TRM cells exert stronger anti-tumor effects and predict better prognosis, which provides ideas for developing new clinically available therapeutic targets.

Fractalkine/CX3CL1 and macrophage inflammatory protein- 1β/CCL4 activity in the rat ovary with induced ovarian hyperstimulation.

Fractalkine (CX3CL1) and macrophage inflammatory protein-1β (MIP-1β)/CCL4 play a role in chemotactic activity, immune response, and inflammatory response. We aimed to investigate the effects of fractalkine and MIP-1β in the development of ovarian hyperstimulation syndrome (OHSS) by considering the inflammatory response during ovulation. Two equal groups of 20 immature female rats were created. Given that one of the rats in the group died, the control group was made up of 9 rats. Group 1 (G1) (n=9): Control group; G2 (n=10): OHSS group. Rats in the G2 group were administered 10 IU FSH for 4 days and 30 IU human chorionic gonadotropin on the fifth day. At 34 days old, all rats were sacrificed, and blood and ovarian tissue samples were collected to measure CX3CL1, CX3CL1R, MIP- 1β, tumor necrosis factor-alpha (TNF-α), interleukin (IL-8), hypoxia-inducible factor (HIF-1α), and interferon-gamma (IFN-γ) levels. Immunohistochemical scoring was performed for CX3CL1 and CX3CL1R in other ovarian tissue. Rat and ovary weights and serum CX3CL1, CX3CLR1, HIF-1α, MIP-1β, TNF-α, IFN-γ and IL-8 levels were significantly higher in G2 than in G1. Tissue IL-8, TNF-α, CX3CL1, CX3CLR1, MIP-1β levels and CX3CL1 and CX3CLR1 immunoreactivity scores were significantly higher in G2 than in G1. CX3CL1 and MIP-1β contribute to the pathophysiology of OHSS by playing a role in the development of inflammation.

Quercetin alleviates ulcerative colitis through inhibiting CXCL8-CXCR1/2 axis: a network and transcriptome analysis

IntroductionUlcerative colitis (UC) is a chronic inflammatory condition of the intestinal tract in which mucosal healing is a crucial measure of therapeutic efficacy. Quercetin, a flavonoid prevalent in various foods and traditional Chinese medicines, exhibits notable pharmacological properties, including antioxidant and anti-inflammatory activities. Consequently, it warrants investigation to determine its potential therapeutic effects on UC. The objective of this study was to investigate the effects and underlying mechanisms of quercetin in a murine model of UC.MethodsA comprehensive approach integrating network predictions with transcriptomic analyses was employed to identify the potential targets and enriched pathways associated with quercetin in UC. Subsequently, the effects of quercetin on pathological morphology, inflammatory mediators, and mucosal barrier-associated proteins, as well as the identified potential targets and enriched pathways, were systematically investigated in a murine model of dextran sulfate sodium (DSS)-induced UC.ResultsNetwork analyses identified CXCL8 and its receptors, CXCR1 and CXCR2, as primary target genes for therapeutic intervention in UC. Further validation through transcriptomic analysis and immunofluorescence staining demonstrated significant upregulation of the CXCL8-CXCR1/2 axis in the intestinal tissues of patients with UC. Experimental investigations in animal models have shown that quercetin markedly alleviates DSS-induced symptoms in mice. This effect includes the restoration of colonic crypt architecture, normalization of goblet cell structure and density, reduction of inflammatory cell infiltration, and decreased concentrations of inflammatory mediators. Quercetin enhanced the expression of tight junction (TJ) proteins, including ZO-1, MUC2 (Mucin 2), and occludin, thereby preserving the integrity of the intestinal mucosal barrier. Additionally, it significantly diminished the levels of IL-17A, NF-κB, CXCL8, CXCR1, and CXCR2 in the colonic tissues of mice with UC.DiscussionThe ameliorative effects of quercetin on colon tissue damage in DSS-induced UC mice were significant, possibly due to its ability to inhibit the CXCL8-CXCR1/2 signaling axis. These findings provide a solid foundation for the clinical application and pharmaceutical advancement of quercetin.

Performance evaluation of CXCL10 ELISA “cosmic” kit to measure CXCL10 in cerebrospinal fluid of patients with HTLV‐1‐associated myelopathy

AbstractObjectivesThis study aimed to validate the clinical utility of cerebrospinal fluid (CSF) CXCL10 measurements in HTLV‐1‐associated myelopathy (HAM) using a CXCL10 ELISA “Cosmic” kit, a more widely applicable method than cytometric bead array (CBA).MethodsCSF CXCL10 levels were measured in 165 samples from 111 patients with HAM and 18 controls using a CXCL10 ELISA “Cosmic” kit. We assessed the following: (1) CSF CXCL10 concentrations by HAM activity level (high, moderate, and low) versus controls; (2) correlation with CBA; (3) cutoff values, sensitivity, and specificity for differentiating among HAM activity levels; (4) changes in HAM activity after steroid therapy; and (5) relationship between HAM activity and prognosis in patients undergoing steroid therapy. A correlation coefficient of ≥0.9 with CBA was the primary endpoint.ResultsThe median CSF CXCL10 levels in the high, moderate, low, and control groups were 4016.0, 841.0, 112.8, and 102.5 pg/mL, respectively. The ELISA findings were highly correlated with the CBA findings (r = 0.99). Cutoff values were set at 2500 pg/mL (sensitivity, 93.3%; specificity, 100%) to distinguish between high and moderate activity and 180 pg/mL (sensitivity, 81.8%; specificity, 100%) for low to moderate activity comparable to CBA. The new cutoffs enabled the detection of HAM activity changes and prediction of motor disability progression under steroid therapy.ConclusionCXCL10 ELISA “Cosmic” kit findings were strongly correlated with CBA findings, meeting the primary endpoint and demonstrating comparable sensitivity and specificity for distinguishing HAM activity. This product shows a promising ability to determine the therapeutic strategy.