Serum Levels Of CXCL10 Research Articles

Malassezia-derived aryl hydrocarbon receptor ligands enhance the CCL20/Th17/soluble CD163 pathogenic axis in extra-mammary Paget's disease.

Malassezia yeast play a role in the pathogenesis of chronic dermatitis, especially in apocrine areas, by polarizing the local immunologic background to a Th2/Th17 state through aryl hydrocarbon receptor (AhR)-dependent pathways. Extra-mammary Paget's disease (EMPD) is an adenocarcinoma of apocrine origin, and except for cases associated with Malassezia yeast and their metabolites, the lesions typically develop in areas not exposed to environmental material. The purpose of this study was to investigate (a) the immunomodulatory effects of Malassezia metabolites on normal human keratinocytes (NHKCs), focusing on interleukin (IL)-17 and related cytokines/chemokines (IL-23, IL-36γ, CCL20), (b) the expression of these factors in lesion-affected skin in EMPD and (c) the activation of tumor-associated macrophages (TAMs) by these factors. Malassezia metabolites augmented the expression of cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1), CCL20 and IL-36γ mRNA in NHKCs in vitro. In lesion-affected skin of patients with EMPD, epidermal keratinocytes expressed CYP1A1 and CCL20. In addition, Paget cells expressed CCL20 and IL-23. IL-17-producing cells were distributed adjacent to Paget cells. Compared to healthy donors, patients with EMPD exhibited significantly increased serum levels of soluble (s)CD163, CXCL5, CXCL10 and CCL20. In addition, serum levels of sCD163 decreased significantly following tumor resection. Our study demonstrates a possible mechanism for the development of EMPD involving AhR-mediated signalling by epidermal keratinocytes and RANKL-induced recruitment of Th17 cells and TAMs.

1736-P: Significance of Anti-insulin Immune Responses in Autoimmune Diabetes with HLA DR9

Previously we reported that CXCL10 in type 1 diabetes (T1D) is higher than controls, and the level reflects islet associated antigen specific T cell responses and disease activity in T1D (Diabetes Care, 2001). However, little is known regarding the relationship between CXCL10 level and autoantibody levels in T1D. Therefore, we investigated relationship between the marker of cellular immunity including CXCL10 levels and islet associated autoantibody levels in T1D. We obtained informed consent and blood samples from 101 T1D patients, and examined HLA (DRB1*04:05, *09:01; susceptible HLA types in Japanese), serum CXCL10, insulin peptide (B9-23, 10-24, 11-25, 12-26) and PPD reactive T cell responses by using ELISPOT (IFNg), GAD, IA-2, ZnT8 Abs and insulin antibody. Because it has been reported that BCG administration might affect the autoimmune responses in T1D, we assessed the PPD responses in these patients as well. Among them, we focused to GAD Ab positive T1D including acute onset (n=28, 13 male and 15 female, mean age 43.1 years, disease duration 5.9 years), and slowly progressive type (n=17, 9 male and 8 female, mean age 56.9 years, disease duration 9.1 years); i.e., autoimmune diabetes subjects. We found significant positive correlation between serum CXCL10 and insulin antibody levels in the autoimmune diabetes with DRB1*09:01 (p<0.05), but it was not the case in those with DRB1*04:05. Interestingly, there was significant positive correlation between insulin peptide reactive T cells and PPD responses in the autoimmune diabetes with DRB1 09:01 (p<0.05), but it was not the case in those with DRB1 04:05. The observation was still true when we focused to acute onset subjects. These results suggest that insulin antibody levels seem to reflect higher disease activity in the autoimmune diabetes with DR9, and modulation of PPD responses may affect insulin peptide T cell responses in those patients. Disclosure A. Shimada: Advisory Panel; Self; Terumo Medical Corporation. Speaker's Bureau; Self; Abbott, Eli Lilly and Company, Novo Nordisk A/S, Sanofi-Aventis. A. Satomura: None. A. Haisa: None. Y. Oikawa: None.

Chemokine CXCL13 as a New Systemic Biomarker for B-Cell Involvement in Acute T Cell-Mediated Kidney Allograft Rejection.

The presence of B-cell clusters in allogenic T cell-mediated rejection (TCMR) of kidney allografts is linked to more severe disease entities. In this study we characterized B-cell infiltrates in patients with TCMR and examined the role of serum CXCL-13 in these patients and experimentally. CXCL-13 serum levels were analyzed in 73 kidney allograft recipients at the time of allograft biopsy. In addition, four patients were evaluated for CXCL13 levels during the first week after transplantation. ELISA was done to measure CXCL-13 serum levels. For further mechanistic understanding, a translational allogenic kidney transplant (ktx) mouse model for TCMR was studied in BalbC recipients of fully mismatched transplants with C57BL/6 donor kidneys. CXCL-13 serum levels were measured longitudinally, CD20 and CD3 composition and CXCL13 mRNA in tissue were examined by flow cytometry and kidneys were examined by histology and immunohistochemistry. We found significantly higher serum levels of the B-cell chemoattractant CXCL13 in patients with TCMR compared to controls and patients with borderline TCMR. Moreover, in patients with acute rejection within the first week after ktx, a >5-fold CXCL13 increase was measured and correlated with B-cell infiltrates in the biopsies. In line with the clinical findings, TCMR in mice correlated with increased systemic serum-CXCL13 levels. Moreover, renal allografts had significantly higher CXCL13 mRNA expression than isogenic controls and showed interstitial CD20+ B-cell clusters and CD3+ cell infiltrates accumulating in the vicinity of renal vessels. CXCL13 blood levels correlate with B-cell involvement in TCMR and might help to identify patients at risk of a more severe clinical course of rejection.

An unusual presentation of purine nucleoside phosphorylase deficiency mimicking systemic juvenile idiopathic arthritis complicated by macrophage activation syndrome

BackgroundSystemic juvenile idiopathic arthritis (sJIA) is an inflammatory condition that presents with fever, rash and arthritis. At onset systemic features are predominant and the diagnosis may be a challenge. Secondary hemophagocytic lymphohistiocytosis (sHLH) forms may be associated with different disorders, including rheumatic diseases, and this form is called macrophage activation syndrome (MAS). CXCL9 levels, a chemokine induced by IFNγ, are significantly elevated in patients with sHLH or MAS and are correlated with laboratory features of disease activity. High levels of IL-18 have been reported in patients with MAS during sJIA, as well as in some patients with sHLH and IL-18 is indeed known to induce IFNγ production.FindingsWe report a patient with a clinical presentation highly suggestive for systemic juvenile idiopathic arthritis (sJIA) onset complicated by MAS, and was later diagnosed with purine nucleoside phosphorylase (PNP)-deficiency with HLH. Some unusual features appeared when HLH was controlled and further investigations provided the correct diagnosis. Serum CXCL9 and IL-18 levels were found markedly elevated at disease onset, during the active phase of MAS and decreased progressively during the course.ConclusionThe reported case underlines the potential difficulties in discriminating sJIA from other causes of systemic inflammation. Furthermore, this supports the notion that especially in young children with a sJIA-like disease other mimicking conditions should be actively sought for. CXCL9 and IL-18 levels suggested that patients with PNP-deficiency may have a subclinical activation of the IFNγ pathway and indeed they are predisposed to develop sHLH.

Potential Effects of CXCL9 and CCL20 on Cardiac Fibrosis in Patients with Myocardial Infarction and Isoproterenol-Treated Rats.

The chemokines CXCL9 and CCL20 have been reported to be associated with ventricular dysfunction. This study was aimed to investigate the effects of CXCL9/CCL20 on cardiac fibrosis following myocardial infarction (MI). Blood samples of patients with MI were obtained to determine the serum CXCL9, CCL20, tumor necrosis factor-α (TNF-α), and transforming growth factor-β (TGF-β). The expression of CXCL9 and CCL20 in hypoxia-incubated H9c2 cells and TNF-α/TGF-β-activated peripheral blood mononuclear cells (PBMCs) were examined. The experimental MI of rats was produced by the intraperitoneal injection of isoproterenol (ISO) (85 mg/kg/day) for two consecutive days. The growth and migration of CXCL9/CCL20-incubated cardiac fibroblasts in vitro were evaluated. TNF-α/TGF-β-activated PBMCs showed an enhanced expression of CXCL9 and CCL20, while hypoxic H9c2 cells did not. Patients with MI had significantly enhanced levels of serum TGF-β and CXCL9 compared to healthy subjects. ISO-treated rats had increased serum CXCL9 levels and marked cardiac fibrosis compared to control rats. The trend of increased serum CCL20 in patients with MI and ISO-treated rats was not significant. CXCL9-incubated cardiac fibroblasts showed enhanced proliferation and migration. The findings of this study suggest that an enhanced expression of CXCL9 following MI might play a role in post-MI cardiac fibrosis by activating cardiac fibroblasts.

Loss of SMAD4 Promotes Colorectal Cancer Progression by Recruiting Tumor-Associated Neutrophils via the CXCL1/8-CXCR2 Axis.

SMAD4 is a key transcriptional factor of TGFβ signaling and acts as a tumor suppressor in colorectal cancer. In the present study, we explored the immunologic effect of SMAD4 on the tumor microenvironment. Using 99 clinical specimens and human colorectal cancer cell lines, we investigate the relationship between SMAD4 expression and neutrophil accumulation. We immunohistochemically analyzed expression of SMAD4, CXCL1, CXCL8, CXCR2, and other proteins with clinical specimens. Finally, we determined the serum levels of CXCL1 and CXCL8 in 125 patients with colorectal cancer. SMAD4 knockdown from human colorectal cancer cells upregulated the expression of CXCL1 and CXCL8, which recruited neutrophils to colorectal cancer tumor via CXCR2. In turn, when neutrophils were exposed to the supernatant of SMAD4-negative colorectal cancer cells, they produced a large amount of CXCL1 and CXCL8 by themselves in vitro. In human clinical specimens, we found that neutrophil infiltration into the peritumoral stroma was more marked in SMAD4-negative colorectal cancer compared with that in SMAD4-positive colorectal cancer, and that both CXCL1 and CXCL8 were abundantly expressed in the tumor-infiltrating neutrophils. Neutrophils isolated from primary colorectal cancer expressed significantly higher levels of CXCL1 and CXCL8 than did those isolated from peripheral blood. Furthermore, tumor-infiltrating neutrophils expressed MMP2 and MMP9 in addition to ARG1 and IDO. Serum CXCL8 level was significantly higher in colorectal cancer patients, especially those at stage II/III, and statistical analysis indicated a high CXCL8 level was associated with a shorter overall survival and relapse-free survival. Blockade of the CXCL1/8-CXCR2 axis could be a novel therapeutic approach against SMAD4-negative colorectal cancer.

Association of Baseline Serum Levels of CXCL5 With the Efficacy of Nivolumab in Advanced Melanoma.

Anti-programmed cell death protein 1 (PD1) antibodies are in wide use for the treatment of various cancers. PD1 antibody-based immunotherapy, co-administration of nivolumab and ipilimumab, is one of the optimal immunotherapies, especially in advanced melanoma with high tumor mutation burden. Since this combined therapy leads to a high frequency of serious immune-related adverse events (irAEs) in patients with advanced melanoma, biomarkers are needed to evaluate nivolumab efficacy to avoid serious irAEs caused by ipilimumab. This study analyzed baseline serum levels of CXCL5, CXCL10, and CCL22 in 46 cases of advanced cutaneous melanoma treated with nivolumab. Baseline serum levels of CXCL5 were significantly higher in responders than in non-responders. In contrast, there were no significant differences in baseline serum levels of CXCL10 and CCL22 between responders and non-responders. These results suggest that baseline serum levels of CXCL5 may be useful as a biomarker for identifying patients with advanced cutaneous melanoma most likely to benefit from anti-melanoma immunotherapy.

The role of high serum CXCL13 level in Waldenström macroglobulinemia

PurposeTo explore the value of elevated CXCL13 levels in Waldenström macroglobulinemia (WM). MethodsWe collected serum samples from 41 patients and bone marrow tissues from 14 patients with newly diagnosed symptomatic WM. Serum and bone marrow samples from patients with other indolent B-cell lymphomas and MGUS were also collected for comparison. Serum CXCL13 levels were measured by enzyme-linked immunosorbent assay, and bone marrow tissues were examined by immunohistochemistry. ResultsThe median serum level of CXCL13 in patients with symptomatic WM was 2483.0 (range 36.8–5644.0) pg/ml, which was significantly higher than in patients with other indolent B-cell lymphomas with monoclonal IgM (median 380.9 pg/ml, range 23.8–5518.0 pg/ml) (p = 0.01). Serum CXCL13 >3250 pg/ml and serum M-protein >38 g/l diagnosed WM with a sensitivity of 98% and specificity of 85%. Serum CXCL13 was strongly correlated with hemoglobin levels (ρ=−0.46, p = 0.002), serum M-protein (ρ=0.47, p = 0.002), and IgM levels (ρ=0.30, p = 0.05) in patients with symptomatic WM. Immunohistochemistry analysis indicated that CXCL13 and activated mast cell levels were also higher in the bone marrow of WM patients compared to patients with IgM-MGUS or other indolent B-cell lymphomas with monoclonal IgM. ConclusionsSerum CXCL13 levels were significantly elevated in patients with WM and correlated with tumor load. Detection of serum CXCL13 may therefore be helpful in the differential diagnosis of WM.

OR19-6 A Novel Anti-CD40 Monoclonal Antibody, Iscalimab, Successfully Treats Graves’ Hyperthyroidism

Objective: Graves’ disease (GD) is an autoimmune disorder characterized by thyrotropin receptor autoantibody (TSHR-Ab)-mediated pathology, leading to hyperthyroidism. To date there is no pharmacotherapy that addresses the underlying pathogenesis. The CD40-CD154 (CD40L) co-stimulatory pathway plays an important role in the pathogenesis of GD by promoting auto-reactive B cell activation and intrathyroidal ectopic lymphoid structure formation and function. Iscalimab (CFZ533) is a fully human, blocking and non-depleting anti-CD40 monoclonal antibody. The purpose of this study was to investigate the efficacy and safety of Iscalimab in patients with Graves’ hyperthyroidism. Methods: In total, fifteen patients with Graves’ hyperthyroidism were enrolled in an open label Phase I proof-of concept study and all completed the trial. Each patient received five doses of Iscalimab at 10 mg/kg IV over 12 weeks. We assessed the effects of Iscalimab on thyroid-related hormones (baseline TSH, free and total T3/T4) and autoantibodies (TSHR-Ab) over 36 weeks following the first dose of Iscalimab. Safety, pharmacokinetics, soluble CD40 levels in plasma, free CD40 on whole blood CD19-positive B cells, and CXCL13 were also assessed. Results: Based on the CD40 occupancy and plasma levels of soluble CD40, the intervention resulted in full CD40 engagement for up to 20 weeks (12-week treatment + 8 week follow up). A clinical response and biochemical euthyroidism was observed in seven out of 15 (46.7%) by Day 211. Free T3/T4 normalized in seven (46.7%) patients who did not received any rescue medication with anti-thyroid drugs. Two further patients (13.3%) did not normalize but did not require rescue medication. In contrast, six (40%) patients required rescue medication with anti-thyroid drugs by Day 141. No patients achieved normalization of TSH by Day 85, however, two out of 15 patients (13.3%) showed normal TSH by Day 169. The levels of free thyroid hormones and TSHR-Ab levels declined over time up to Day 211. The mean levels of TSHR-Ab declined from 15.3 IU/L (baseline) to 9.4 IU/L at Day 85 (35% reduction, P<0.001), then further decreased to 4.0 IU/L at day 141 (66% reduction, P<0.001). The TSHR-Ab levels fell in normal range in four out of 15 patients (26.7%). Treatment with Iscalimab was associated with a rapid and significant reduction in the serum levels of CXCL13 (94.1 pg/mL at baseline to 68.5 pg/mL at Day 15) and remained at similar levels during the 12 week treatment period. Twelve patients reported at least one adverse event. Most events were mild in nature (34 in 8 patients, 53.3%) and few were moderate (5 in 4 patients, 26.7%). Conclusions: Iscalimab was generally safe and clinically effective in a subgroup of patients with Graves’ hyperthyroidism. These encouraging results suggest that Iscalimab may be an attractive strategy as the immunomodulation treatment for GD.

Clinical significance of serum CXCL9 levels as a biomarker for systemic juvenile idiopathic arthritis associated macrophage activation syndrome

To clarify cytokines involved in the development of systemic juvenile idiopathic arthritis (s-JIA) associated macrophage activation syndrome (MAS) and to identify the serum biomarkers for the diagnosis of s-JIA associated MAS, we employed an antibody array that simultaneously detects 174 cytokines. Fifteen s-JIA patients including 5 patients receiving tocilizumab (TCZ) were analyzed. The levels of five cytokines were significantly elevated in MAS phase compared to those in the active phase of s-JIA. CXCL9 showed the most significant increase following the development of s-JIA associated MAS. Next, to confirm clinical significance of serum CXCL9 levels as a biomarker for s-JIA associated MAS, serum CXCL9 levels in 56 patients with s-JIA including 20 with MAS were analyzed. Results were compared with the clinical features of s-JIA associated MAS. Serum CXCL9 levels correlated positively with disease activity. Monitoring of serum CXCL9 is useful for the evaluation of disease activity in s-JIA associated MAS.

Circulating levels of CXCL11 and CXCL12 are biomarkers of cirrhosis in patients with chronic hepatitis C infection

Background & aimsThe chemokines CXCL10 (interferon ϒ-inducible protein 10 [IP-10]), CXCL11 (Human interferon inducible T cell alpha chemokine [I-TAC]), and CXCL12 (stromal cell derived factor 1 [SDF-1]) contribute to cell recruitment, migration, activation, and homing in liver diseases and their serum levels have been shown to be associated with the degree of liver inflammation or fibrosis in various etiologies. However, the data may be contradictory or insufficient, particularly for CXCL12, in the field of chronic HCV infection. Here, we aimed to provide evidence for these chemokines as biomarkers for chronic HCV infection. MethodsWe analyzed the serum concentration of the three chemokines in healthy donors (n = 39) and patients (n = 87) with chronic HCV infection. Chemokine serum levels were compared to the stage of liver inflammation and fibrosis obtained from liver biopsies. ResultsSerum CXCL10 and CXCL11 levels were higher at advanced stages of liver inflammation than at earlier stages, but the results were only of medium significance. Both serum CXCL11 and CXCL12 levels were significantly higher in cirrhotic patients than those with low or medium stages of fibrosis. The AUROCs were 0.8167 and 0.8574, respectively, for the diagnosis of cirrhotic patients. ConclusionThese data provide evidence for the value of CXCL10, CXCL11, and CXCL12 as biomarkers of liver inflammation and fibrosis during chronic HCV infection. Serum CXCL10 and CXCL11 levels were associated with liver inflammation, but the level of significance was insufficient. However, serum CXCL11 and CXCL12 levels were elevated in cirrhotic patients, showing equivalent diagnostic accuracy as the existing established single serum fibrosis markers or algorithms.

CXCL13/CXCR5 Axis Predicts Poor Prognosis and Promotes Progression Through PI3K/AKT/mTOR Pathway in Clear Cell Renal Cell Carcinoma.

The chemokine ligands and their receptors play critical roles in cancer progression and patients outcomes. We found that CXCL13 was significantly upregulated in ccRCC tissues compared with normal tissues in both The Cancer Genome Atlas (TCGA) cohort and a validated cohort of 90 pairs ccRCC tissues. Statistical analysis showed that high CXCL13 expression related to advanced disease stage and poor prognosis in ccRCC. We also revealed that serum CXCL13 levels in ccRCC patients (n = 50) were significantly higher than in healthy controls (n = 40). Receiver operating characteristic (ROC) curve revealed that tissue and serum CXCL13 expression might be a diagnostic biomarker for ccRCC with an area under curve (AUC) of 0.809 and 0.704, respectively. CXCL13 was significantly associated with its receptor, CXCR5, in ccRCC tissues, and ccRCC patients in high CXCL13 high CXCR5 expression group have a worst prognosis. Functional and mechanistic study revealed that CXCL13 promoted the proliferation and migration of ccRCC cells by binding to CXCR5 and activated PI3K/AKT/mTOR signaling pathway. These results suggested that CXCL13/CXCR5 axis played a significant role in ccRCC and might be a therapeutic target and prognostic biomarker.

Increased Circulating CXCL10 in Non-Segmental Vitiligo Concomitant with Autoimmune Thyroid Disease and Alopecia Areata.

BackgroundVitiligo is a common acquired pigmentary disease caused by destruction of epidermal melanocytes in underlying autoimmune response. Few studies have been focused on the role of chemokines in non-segmental vitiligo (NSV) concomitant with autoimmune thyroid disease (AITD) and alopecia areata (AA).ObjectiveThe aim of this study was to determine the best serum biomarker for predictive role in the progression of vitiligo and to evaluate the influence of AA and/or AITD on vitiligo by using the biomarker.MethodsThis prospective cohort study recruited 45 NSV patients: 14 without either AITD or AA, 12 with AITD, 11 with AA, and 8 with both AITD and AA. Serum levels of CXCL1, CXCL8, CXCL9, CXCL10, CXCL12, CXCL13, and CXCL16 were analyzed by ELISA. CXCR3 mRNA expression was detected on PBMCs by RT-PCR. Improvement was evaluated using repigmentation scales.ResultsSerum CXCL10 levels, along with the expression of CXCR3 mRNA were higher in NSV patients with AITD or AA alone than in those without AITD or AA. Moreover, serum CXCL10 levels, along with the expression of CXCR3 mRNA were higher in NSV patients with both AITD and AA than in those with AITD or AA alone. Poorer repigmentation was observed in NSV patients with both AA and AITD than in those with AA or AITD alone.ConclusionCXCL10 could be a biomarker to predict the progression of NSV. Dermatologists should pay much attention to those NSV patients concomitant with AITD and/or AA, for comorbidity might lead to more active autoimmune reaction.

Factors regulating the activity of b-lymphocytes, as potential biomarkers of multiple sclerosis

To assess the possibility of using serum levels of chemokine CXCL13, B-cell activating factor (BAFF), interleukin 1 and 17 (IL-10, IL-17) as markers of the development/progression of multiple sclerosis (MS). Sixty-seven patients with MS in remission and 14 healthy volunteers were examined. The levels of BAFF, CXCL13 and the main products of Th17 and Treg - interleukin 17 (IL-17) and IL-10 in the serum were determined, their relationship with clinical characteristics of MS during remission was assessed. The content of IL-17 and BAFF in the serum of patients with MS did not differ significantly from the control group, and the levels of IL-10 and CXCL13 were lower than the corresponding indicators of healthy donors. In patients with MS, no correlation was found between IL-17, IL-10, BAFF and clinical characteristics of MS. The level of CXCL13 correlated with quality of life, fatigue, clinical and functional status of patients. According to the results of this study, BAFF and CXCL13 cannot be proposed as diagnostic biomarkers of MS.

Association of metabolic syndrome with inflammatory markers in a sample of community-dwelling older adults.

The study aimed to identify the cutoff points for inflammatory markers that best discriminate the occurrence of metabolic syndrome in community-dwelling older adults. Baseline data were used from the elderly cohort in the city of Bambuí, Minas Gerais State, Brazil. The target exposure was presence of metabolic syndrome, defined according to the Adult Treatment Panel III criterion, and the outcomes included the following inflammatory markers: cytokines (IL-1β, IL-6, IL-10, IL-12 e TNF), chemokines (CXCL8, CXCL9, CCL2, CXCL10, and CCL5), and C-reactive protein (CRP). Definition of the cutoff points for the inflammatory markers was based on the Classification and Regression Tree (CART) method. The associations between these markers and metabolic syndrome were estimated by logistic regression models, obtaining odds ratios and 95% confidence intervals, considering adjustment for confounding factors. Prevalence of metabolic syndrome was 49.1%, and IL-1β, IL-12, and TNF levels were not associated statistically with this exposure. After adjustment, presence of metabolic syndrome was associated with higher IL-6 and CRP levels and lower CXCL8 and CCL5. Significant associations were also observed with intermediate serum CXCL9 and CXCL10 levels. The combination of markers also showed a significant and consistent association with metabolic syndrome. In addition to demonstrating an association between metabolic syndrome and a wide range of biomarkers (some not previously described in the literature), the results highlight that this association occurs at much lower levels than previously demonstrated, suggesting that metabolic syndrome plays an important role in the inflammatory profile of the older adults.

Stromal-derived factor-1 (CXCL12) and its receptor (CXCR4) in neonatal sepsis

Background Neonatal sepsis remains a major cause of morbidity and mortality in newborns. The C-X-C motif chemokine 12 (CXCL12) and its receptor α-chemokine receptor type 4 (CXCR4) are now known to play an important role in inflammatory states and mediate lymphocyte migration in response inflammation. However, it is unclear how chemokines respond to late-onset neonatal sepsis (LOS). Objective The aim of this study was to assess the value of measuring the serum levels of stromal-derived factor-1, also named CXCL12, and lymphocyte expression levels of its receptor CXCR4 in the diagnosis of LOS in relation to clinical and hematological sepsis scores. Patients and methods Levels of CXCL12 in serum and lymphocyte expression of CXCR4 were determined using enzyme-linked immunosorbent assay technique and flow cytometry, respectively, in 38 full-term neonates; 23 cases of LOS (13 male and 10 female) and 15 healthy neonates (six male and nine female) were included in the study. Results Serum levels of CXCL12 and lymphocyte expression levels of CXCR4 were significantly higher in neonates with LOS when compared with nonseptic neonates. At a cutoff point of 30 pg/ml, the diagnostic accuracy of CXCL12 in the diagnosis of neonatal sepsis was 100%, with a sensitivity of 100% and a specificity of 100%, whereas the diagnostic accuracy of mean fluorescence intensity of lymphocyte expression of CXCR4 in the diagnosis of neonatal sepsis was 84%, with a sensitivity of 87%, specificity of 80%, positive predictive value of 87%, and negative predictive value of 80% at a cutoff point of 150. Conclusion CXCR4 and CXCL12 levels increase significantly in septic neonates and they are valuable markers in the diagnosis of neonatal sepsis.

The temporal pattern of immune and inflammatory proteins prior to a recurrent coronary event in post-acute coronary syndrome patients

Purpose: We assessed the temporal pattern of 29 immune and inflammatory proteins in post-acute coronary syndrome (ACS) patients, prior to the development of recurrent ACS.Methods: High-frequency blood sampling was performed in 844 patients admitted for ACS during one-year follow-up. We conducted a case-control study on the 45 patients who experienced reACS (cases) and two matched event-free patients (controls) per case. Olink Proteomics’ immunoassay was used to obtain serum levels of the 29 proteins, expressed in an arbitrary unit on the log2-scale (Normalized Protein eXpression, NPX). Linear mixed-effects models were applied to examine the temporal pattern of the proteins, and to illustrate differences between cases and controls.Results: Mean age was 66 ± 12 years and 80% were men. Cases and controls had similar baseline clinical characteristics. During the first 30 days, and after multiple testing correction, cases had significantly higher serum levels of CXCL1 (difference of 1.00 NPX, p = 0.002), CD84 (difference of 0.64 NPX, p = 0.002) and TNFRSF10A (difference of 0.41 NPX, p < 0.001) than controls. After 30 days, serum levels of all 29 proteins were similar in cases and controls. In particular, no increase was observed prior to reACS.Conclusions: Among 29 immune and inflammatory proteins, CXCL1, CD84 and TNFRSF10A were associated with early reACS after initial ACS-admission.

Myo-inositol in autoimmune thyroiditis, and hypothyroidism.

Myo-inositol (Myo-Ins) plays an important role in thyroid function and autoimmunity. Myo-Ins is the precursor for the synthesis of phosphoinositides, which takes part in the phosphatidylinositol (PtdIns) signal transduction pathway, and plays a decisive role in several cellular processes. In the thyroid cells, PtdIns is involved in the intracellular thyroid-stimulating hormone (TSH) signaling, via Phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P3) (PIP-3). Moreover, the phosphatidyl inositol 3 kinases (PI3K) family of lipid kinases regulates diverse aspects of T, B, and Tregs lymphocyte behaviour. Different mouse models deficient for the molecules involved in the PIP3 pathway suggest that impairment of PIP3 signaling leads to dysregulation of immune responses and, sometimes, autoimmunity. Studies have shown that cytokines modulate Myo-Ins in thyroid cells. Moreover, clinical studies have shown that after treatment with Myo-inositol plus seleniomethionine (Myo-Ins + Se), TSH levels significantly declined in patients with subclinical hypothyroidism due to autoimmune thyroiditis. The treatment was accompanied by a decline ofantithyroid autoantibodies. After treatment serum CXCL10 levels declined, confirming the immune-modulatory effect of Myo-Ins. Additional research is necessary in larger population to evaluate the effect on the quality of life, and to study the mechanism of the effect on chemokines.

Genetic Risk of Severe Chronic Graft-Versus-Host Disease Defined By Host-Derived CXCR3 Ligands

IntroductionSevere chronic graft-versus-host disease (cGVHD) is the leading cause of morbidity and mortality in long-term survivors after allogeneic stem cell transplantation (alloSCT). The CXCR3 signalling pathway may be implicated in cGVHD pathophysiology, since CXCR3 and its ligands (CXCL4, CXCL9, CXCL10 and CXCL11) are involved in attracting activated Th1 cells into inflamed tissues. To better understand the role of the CXCR3 axis in cGVHD, we measured serum levels of CXCR3 ligands in allograft recipients pre-transplant and on day 28 post-transplant, and correlated them to the single nucleotide polymorphisms (SNPs) in recipient CXCR3/CXCR3L genes in the context of severe cGVHD.Patients and methods:287 patients who were allografted at Heidelberg University Hospital, survived more than 6 months after alloSCT, and did not receive statin-based endothelial prophylaxis (SEP) constituted the no-SEP training cohort, whereas 401 patients who received SEP constituted the SEP cohort. DNA for genotyping was available for 545 patients (no-SEP 242, SEP 303) and sera for measuring CXCL9, CXCL10 and CXCL11 by ELISA were collected at pre-transplant for 405 patients (no-SEP 109, SEP 296) and at day +28 for 494 patients (no-SEP 152, SEP 342). The no-SEP validation cohort consisted of 202 patients who had been allografted at Berlin Charité and survived more than 6 months. CGVHD was diagnosed and graded using the National Institutes of Health's 2005 consensus criteria.Eighteen SNPs (7 in CXCL9-11, 7 in CXCL4 and 4 in CXCR3 loci) were selected and analyzed for association with severe cGVHD, treating non-relapse death and relapse without severe cGVHD as competing events. Associations of SNPs with serum chemokine levels were studied by Mann-Whitney U-test. Hazard ratios (HR) with 95% confidence interval (CI) were estimated using (cause-specific) Cox regression. Covariates considered in multivariate analysis were age, diagnosis, donor type, sex of donor and recipient and usage of ATG.Results:Overall, 50 of 287 patients (17.4%) in the no-SEP training cohort, 53 of 401 patients (13.2%) in the SEP cohort and 48 of 202 patients (23.8%) in the no-SEP validation cohort developed at least one episode of severe cGVHD. In the no-SEP training cohort, higher serum CXCL9 levels at day +28 were significantly associated with a higher risk of severe cGVHD in univariate analysis (HR 1.38 for every log2-fold change, 95% CI 1.10-1.75, P=0.01; Figure 1a). No significant association was found for serum CXCL10 and CXCL11 pre- or post-transplant with severe cGVHD.The rs884304 SNP in CXCL9-11 locus showed a significant association with severe cGVHD; patients with AA/AG genotypes carried a HR of 2.32 (95%CI 1.21-4.46, P=0.01) compared to patients with GG genotypes. In addition, 3 other SNPs (rs3733236 and rs4282209 in CXCL9-11, rs655328 in CXCL4 loci) were selected based on the effect on severe cGVHD (P <0.10) to calculate a combined genetic risk score. Patients with any low-risk genotypes (rs884304GG, rs3733236AA/AG, rs4282209AA/AG and rs655328TT) were classified as the low-risk. All others were considered as high-risk. Taken together, high-risk patients were found in 21.4% (52/242) of the no-SEP training cohort, 22.4% (68/303) of the SEP cohort and 19.8% (40/202) of the no-SEP validation cohort. Patients in the high-risk group had significantly higher serum CXCL9 levels at day +28 (Figure 1b) and a significantly higher risk of severe cGVHD (Figure 1c) on both univariate (HR 2.68, 95%CI 1.45-4.95, P=0.001) and multivariate analyses (HR 2.49, 95%CI 1.33-4.66, P=0.004). The effect of the combined score was confirmed in the no-SEP validation cohort (HR 3.02, 95%CI 1.60-5.72, P=0.001).In contrast, in the SEP cohort the adverse effect of high risk genotypes was not observed (HR 1.30, 95% CI 0.60-2.79, P=0.50). In addition, SEP reduced day +28 CXCL9 levels in patients with high-risk genotype but not in low-risk patients.Conclusion:In the absence of SEP, the risk of severe cGVHD could be predicted both by a genetic score of 4 SNPs in recipient CXCR3L genes and by serum CXCL9 levels at day +28. The genetic score influenced serum CXCL9 levels at day +28. Our results suggest that in high-risk patients, host-derived CXCR3 ligands are upregulated early after alloSCT and may promote the development of severe cGVHD. Endothelial prophylaxis may reduce the risk of severe cGVHD by regulating serum CXCL9 levels and, thus, warrants further study. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

CXCL12 and CXCR4 in the Peripheral Blood of Patients with Parkinson’s Disease

Objective: The role of CXCL12 and its receptor CXCR4 has not been fully examined in Parkinson’s disease (PD). The purpose of this study was to investigate the role of CXCL12/CXCR4 in the peripheral blood of patients with PD and healthy controls. Methods: CXCL12 serum levels and CXCR4 mRNA levels were measured in 30 PD patients and 40 controls using ELISA and real-time PCR, respectively. Results: CXCL12 serum levels were significantly higher in PD patients compared to controls (p < 0.0001). Moreover, CXCR4 expression in peripheral blood mononuclear cells (PBMC) of PD patients was significantly increased compared to controls (p < 0.0001). Conclusions: Our findings provide new information on the expression of CXCL12/CXCR4 in PD. CXCR4 expression in PBMC or CXCL12 serum levels may be potential biomarkers of inflammation in PD patients.