The serum levels of FCGBP, SAA1, CXCL10 in children with mycoplasma pneumoniae pneumonia and their relationship with prognosis / 中国医师杂志

Objective:To investigate the serum levels and clinical significance of Fc fragment of the IgG-binding protein (FCGBP), serum amyloid protein A1 (SAA1), and CXC chemokine ligand 10 (CXCL10) in children with mycoplasma pneumoniae pneumonia (MPP) and their relationship with prognosis.Methods:A prospective study was conducted on 122 children with MPP admitted to the department of pediatrics of the 970th Hospital of the Joint Logistics Support Force of the Chinese People′s Liberation Army from January 2019 to December 2021. According to the severity and prognosis of MPP, they were divided into mild and severe groups, good prognosis group, and poor prognosis group. Forty healthy children who underwent physical examination during the same period were set as the control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum levels of FCGBP, SAA1, and CXCL10 in each subject, and to compare the differences in serum levels of FCGBP, SAA1, and CXCL10 among different groups. Multivariate logistic regression analysis was used to investigate the influencing factors of poor prognosis in MPP patients. The diagnostic value of individual and combined detection of serum procalcitonin (PCT), FCGBP, SAA1, and CXCL10 for poor prognosis in MPP children by analyzing the receiver operating characteristic (ROC) curve.Results:The levels of serum FCGBP [(115.68±10.57)ng/ml, (78.41±6.73)ng/ml, (12.55±3.25)ng/ml], SAA1 [(34.18±3.72)mg/L, (25.54±2.63)mg/L, (6.74±0.82)mg/L], and CXCL10 [(714.26±55.64)ng/L, (353.74±42.67)ng/L, (106.25±12.92)ng/L] in the severe MPP group were significant higher than those in the mild MPP group and the control group, with statistical significance (all P<0.05). The white blood cell (WBC), neutrophil percentage, C reactive protein (CRP), erythrocyte sedimentation rate (ESR), PCT, lactate dehydrogenase (LDH), D-dimer (D-D), FCGBP, SAA1, CXCL10 of the children in the poor prognosis group were significantly higher than those in the good prognosis group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis showed that increased PCT ( OR=1.603, 95% CI: 1.190-2.160), FCGBP ( OR=1.757, 95% CI: 1.115-2.770), SAA1 ( OR=1.900, 95% CI: 1.327-2.720) and CXCL10 ( OR=1.704, 95% CI: 1.212-2.397) were independent risk factors for poor prognosis of MPP children (all P<0.05). The combined detection of serum PCT, FCGBP, SAA1, and CXCL10 had a significantly higher diagnostic value for the risk of poor prognosis in children with MPP than a single indicator. Conclusions:The elevated levels of serum FCGBP, SAA1, and CXCL10 in children with MPP are associated with the severity of MPP and are independent risk factors for poor prognosis in MPP patients.

Construction of interferon alpha/beta receptor subunit 1 gene knockout Caco-2 cell line based on CRISPR/Cas9 system / 中国生物制品学杂志

@#Objective To knockout interferon alpha/beta receptor subunit 1(IFNAR1) gene in human colorectal adenocarcinoma cells Caco-2 using clustered regularly interspaced short palinmic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)system to construct IFNAR1 knockout Caco-2 cell line.Methods The single guide RNA(sgRNA)sequence was designed to specifically recognize the exon region of IFNAR1 gene using CRISPR/Cas9 technology,and the LentiCRISPRv2-IFNAR1-sgRNA recombinant plasmid was constructed.Caco-2 cells were infected with the plasmid packaged by lentivirus and screened by puromycin resistance.The obtained monoclonal cell lines were cultured by limited dilution method,which were verified for the effect of IFNAR1 gene knockout by target gene sequencing and Western blot,and detected for the mRNA levels of CXC chemokine ligand 10(CXCL10)and interferon-stimulatd gene 20(ISG20)in IFNAR1knockout cells by adding exogenous IFNβ.Results Sequencing results of plasmid LentiCRISPRv2-IFNAR1-sgRNA showed that the insertion sites were all located at the sticky end of BsmBⅠenzyme digestion.Two IFNAR1 knockout monoclonal cell lines were obtained.The sequencing results showed that Caco-2-IFNAR1-KO1 had 5 bp deletion in the sixth exon of IFNAR1,and Caco-2-IFNAR1-KO2 had 18 bp deletion and 1 bp insertion in the seventh exon.Compared with wild-type Caco-2 cells,Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells showed no expression of IFNAR1 protein.Compared with no IFNβ stimulation,the mRNA levels of CXCL10 gene(t = 0.566 and 1.268 respectively,P>0.05)and ISG20 gene(t =1.522 and 1.733 respectively,P>0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells stimulated by 50 ng/mL IFNβ showed no significant increase.While compared with those of wild-type Caco-2 cells,the mRNA levels of CXCL10gene(t = 6.763 and 6.777 respectively,P<0.05)and ISG20 gene(t = 5.664 and 5.65 respectively,P<0.05)in Caco-2-IFNAR1-KO1 and Caco-2-IFNAR1-KO2 cells decreased significantly under the stimulation of 50 ng/mL exogenous IFNβ.Conclusion Caco-2 cell line with IFNAR1 knockout was successfully constructed by using CRISPR/Cas9 technology,and the downstream molecules activated by IFNAR(interferon alpha/beta receptor)in this cell line were obviously inhibited,which provided a powerful tool for further exploration of the innate immune response and replication packaging mechanism of Caco-2 cells after virus infection.

Research progress on CXCR3 variants and their ligands in tumor microenvironment / 中国肿瘤生物治疗杂志

@#[摘 要] 恶性肿瘤是严重威胁人类生命的疾病之一，近年来免疫治疗已经成为肿瘤治疗的焦点，解决免疫治疗只对部分患者有效的问题迫在眉睫。在肿瘤微环境（tumor microenvironment，TME）中趋化因子介导细胞的定向移动，同时具有多种调节功能，既可以作用于免疫细胞，也可直接作用于肿瘤细胞，发挥了复杂的生物学作用。CXC趋化因子受体3（CXC chemokine receptor 3，CXCR3）通过与其同源CXC趋化因子配体9（CXC chemokine ligand 9，CXCL9）/10/11结合，不仅参与了肿瘤发生、侵袭并促进肿瘤相关血管的形成，同时也介导了免疫细胞向肿瘤组织中浸润，为无免疫反应性或免疫反应性差的“冷肿瘤”转变为免疫反应性的“热肿瘤”提供了新的思路，并且可能成为治疗的新靶点。这种抗肿瘤和促肿瘤的双重作用，似乎与CXCR3变体（CXCR3-A、CXCR3-B）发挥相反的作用密切相关。本文就近年来CXCR3变体CXCR3-A、CXCR3-B及其配体CXCL9/10/11在TME中作用的研究进展展开综述。

Effect of chemokines CXCL9 and CXCL10 on bone erosion in patients with rheumatoid arthritis / 北京大学学报(医学版)

OBJECTIVE@#To detect the serum level of soluble chemokines CXCL9 and CXCL10 in patients with rheumatoid arthritis (RA), and to analyze their correlation with bone erosion, as well as the clinical significance in RA.@*METHODS@#In the study, 105 cases of RA patients, 90 osteoarthritis (OA) patients and 25 healthy controls in Peking University People's Hospital were included. All the clinical information of the patients was collected, and the serum CXCL9 and CXCL10 levels of both patients and healthy controls were measured by enzyme-linked immune sorbent assay (ELISA). CXCL9 and CXCL10 levels among different groups were compared. The correlation between serum levels with clinical/laboratory parameters and the occurrence of bone erosion in RA were analyzed. Independent sample t test, Chi square test, Mann-Whitney U test, Spearman's rank correlation and Logistic regression were used for statistical analysis.@*RESULTS@#The levels of CXCL9 and CXCL10 were significantly higher in the RA patients [250.02 (126.98, 484.29) ng/L, 108.43 (55.16, 197.17) ng/L] than in the OA patients [165.05 (75.89, 266.37) ng/L, 69.00 (33.25, 104.74) ng/L] and the health controls [79.47 (38.22, 140.63) ng/L, 55.44 (18.76, 95.86) ng/L] (all P < 0.01). Spearman's correlation analysis showed that the level of serum CXCL9 was positively correlated with swollen joints (SJC), rheumatoid factor (RF) and disease activity score 28 (DAS28) (r=0.302, 0.285, 0.289; P=0.009, 0.015, 0.013). The level of serum CXCL10 was positively correlated with tender joints (TJC), SJC, C-reactive protein (CRP), immunoglobulin (Ig) A, IgM, RF, anti-cyclic citrullinated peptide antibody (ACPA), and DAS28 (r=0.339, 0.402, 0.269, 0.266, 0.345, 0.570, 0.540, 0.364; P=0.010, 0.002, 0.043, 0.045, 0.009, < 0.001, < 0.001, 0.006). Serum CXCL9 and CXCL10 levels in the RA patients with bone erosion were extremely higher than those without bone erosion [306.84 (234.02, 460.55) ng/L vs. 149.90 (75.88, 257.72) ng/L, 153.74 (89.50, 209.59) ng/L vs. 54.53 (26.30, 83.69) ng/L, respectively] (all P < 0.01). Logistic regression analysis showed that disease duration, DAS28 and serum level of CXCL9 were correlated with bone erosion in the RA patients (P < 0.05).@*CONCLUSION@#Serum levels of CXCL9 and CXCL10 were remarkably elevated in patients with RA, and correlated with disease activities and occurrence of bone erosion. Chemokines CXCL9 and CXCL10 might be involved in the pathogenesis and bone destruction in RA.

CXCL10/CXCR3 Signaling in the DRG Exacerbates Neuropathic Pain in Mice / 神经科学通报·英文版

Chemokines and receptors have been implicated in the pathogenesis of chronic pain. Here, we report that spinal nerve ligation (SNL) increased CXCR3 expression in dorsal root ganglion (DRG) neurons, and intra-DRG injection of Cxcr3 shRNA attenuated the SNL-induced mechanical allodynia and heat hyperalgesia. SNL also increased the mRNA levels of CXCL9, CXCL10, and CXCL11, whereas only CXCL10 increased the number of action potentials (APs) in DRG neurons. Furthermore, in Cxcr3

Effect of exogenous chemokines on the function of human umbilical vein endothelial cells under the environment of high glucose / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the effects of exogenous CXCL9, CXCL10 and CXCL11 on human umbilical vein endothelial cells under a high glucose environment and explore its mechanisms.<p>METHODS: The cells in logarithmic growth stage were divided into control group(glucose concentration 5.5mmol/L), low glucose group(glucose concentration 5mmol/L), high glucose group(glucose concentration 30mmol/L). CXCL9(100ng/mL), CXCL10(10ng/mL)and CXCL11(100ng/mL)were added respectively, cultured for 24, 48 and 72h. CCK-8 method was used to detect cell proliferation, RT-PCR was used to detect CXCR3 mRNA expression, and immunofluorescence was used to detect Ki-67 expression.<p>RESULTS: The results of CCK-8 method showed that the absorbance value of the control group increased gradually with the increase of time after adding three exogenous chemokines. The absorbance value of the low glucose group increased first and then decreased, reaching the peak at 48h. The absorbance value of the high glucose group decreased generally. The results of RT-PCR showed that the expression of CXCR3 mRNA in low glucose group and high glucose group was higher than that in 24h after adding CXCL9, CXCL10 and CXCL11 for 48 and 72h. The results of immunofluorescence showed that the fluorescence intensity of Ki-67 decreased in the low and high glucose 72h after adding CXCL9, CXCL10 and CXCL11. The change in the high glucose group is more obvious.<p>CONCLUSION: Exogenous CXCL9, CXCL10 and CXCL11 can decrease the activity of human umbilical vein cell under high glucose environments and induce the increase in CXCR3 expression. The increase of CXCR3 reached the highest after adding exogenous CXCL10 and CXCL11, suggesting a target for clinical intervention of diabetic retinopathy.

Correlation of miRNA-181c expression in peripheral blood mononuclear cells with interferon-γ, chemokine (C-X-C motif) ligand 10, and Toll-like receptor 4 in children with autoimmune hepatitis / 临床肝胆病杂志

ObjectiveTo investigate the correlation of miR-181c expression in peripheral blood mononuclear cells (PBMCs) with interferon-γ (IFN-γ), chemokine (C-X-C motif) ligand 10 (CXCL10), and Toll-like receptor 4 (TLR4) in children with autoimmune hepatitis (AIH). MethodsA total of 27 children with AIH who were admitted to The Affiliated Hospital of Yanbian University from March 2015 to May 2019 were enrolled as AIH group, and 30 healthy children who underwent physical examination during the same period of were enrolled as control group. The expression of miR-181c in PBMCs and the expression of IFN-γ, CXCL10, and TLR4 were measured for the two groups. The t-test was used for comparison of normally distributed continuous data between two groups, and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. The Pearson correlation coefficient was used to investigate the correlation of miR-181c expression with each index, and a logistic regression analysis was used to investigate the influence of each factor on AIH. ResultsCompared with the control group, the AIH group had significantly higher levels of the liver function parameters aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), and total bilirubin (TBil) (t=14.445,20.064，11.728,13.822, all P＜0.001). The AIH group also had significantly higher levels of IgA, IgM, and IgG than the control group (t=7.772, 5147, and 6771, all P＜0.05). The AIH group had significantly lower relative expression of miR-181c in PBMCs than the control group (0.784±0173 vs 1.106±0.224, t=5.819, P＜0.05). Compared with the control group, the AIH group had significantly higher levels of IFN-γ and CXCL10 and mRNA expression of TLR4 (t=6.949, 12.303, and 13.835, all P＜0.05). The correlation analysis showed that in the children with AIH, the expression of miR-181c in PBMCs was negatively correlated with IFN-γ, CXCL10, TLR4, AST, ALT, GGT, TBil, and IgG (r=-0.316, -0.348, -0.322, -0.427, -0.442, -0.408, -0.396, and -0.321, all P＜0.05). The univariate logistic regression analysis showed that AST, ALT, GGT, TBil, IFN-γ, CXCL10, TLR4 mRNA, and miR-181c were all included in the regression model (all P＜0.05) and were the influencing factors for the onset of AIH. ConclusionChildren with AIH have downregulated expression of miR-181c in PBMCs, which is closely associated with IFN-γ, CXCL10, and TLR4, suggesting that miR-181c may affect the development of AIH in children by regulating the immune system.

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

BACKGROUND: Vitiligo 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). OBJECTIVE: The 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. METHODS: This 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. RESULTS: Serum 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. CONCLUSION: CXCL10 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.

LncRNA53106 regulates CXCL10 and affects the apoptosis of islet β cells / 中华内分泌代谢杂志

Objective@#To investigate the regulatory mechanism of long non-coding RNA (lncRNA) 53106 in the apoptosis model of MIN6 cells stimulated by cytokines.@*Methods@#The stimulation model of cytokines 10 ng/ml interleukin-1β, 50 ng/ml tumor necrosis factor-α, 50 ng/ml interferon-γ in MIN6 islet cell lines were established. The apoptosis rate was measured by flow cytometry and the expression levels of lncRNA53106 and (C-X-C motif) ligand (CXCL)10 were detected by realtime quantitative PCR (qPCR). LncRNA53106 smart silencer and CXCL10 siRNA were constructed, and lncRNA53106 and CXCL10 were knocked down respectively. Then inflammatory cytokines were combined to stimulate, and their roles in the apoptosis of min6 cells were detected by flow cytometry, qPCR, and Western blotting.@*Results@#In the apoptosis model of MIN6 cells stimulated by cytokines, the apoptosis rate of cytokines group was significantly increased and reached statistical significance. The apoptosis rate of the knocked down lncRNA53106 group was significantly lower than that of the control group (P<0.05). The expression of CXCL10 was also decreased in the knockdown group by qPCR and Western blotting, the expressions of the apoptosis-related factors Bax and Caspase3 mRNA were decreased. The apoptosis rate in the knocked down CXCL10 group was significantly lower than that in the control group (P<0.05), and the expression of lncRNA53106 was slightly increased, but the difference was not significant (P=0.61).@*Conclusion@#LncRNA53106 may promote the expression of apoptosis factor by upregulation of CXCL10, and promote the apoptosis of β cells of the pancreas, which may lead to the occurrence of type 1 diabetes.

Effects of tacrolimus in treating vitiligo on the secretion of local chemokine / 中华医学美学美容杂志

Objective To analyze the abnormalities of local chemokines in patients with vitiligo and to explore the effect of tacrolimus on the secretion of chemokines in keratinocytes.Methods Blister fluids of 50 patients with vitiligo were collected,including lesion areas and normal areas.Luminex was used to analyze the concentration of local chemokines in patients with vitiligo to determine whether the chemokines were closely related to vitiligo.The effect of tacrolimus on chemokine secretion of was analyzed by Western blot in HaCaT cells.Results By Luminex analysis of blister fluid,it was found that CXCL9 and CXCL10 were significantly higher in the leukoplakia of vitiligo,and there was a significant difference,compared with the blister fluid in the normal site (P＜0.01).IFN-γ significantly stimulated the keratinocyte cell line HaCat to express CXCL9 and CXCL10.After pretreatment of HaCaT cells with 20 mg tacrolimus,the expression of CXCL9 and CXCL10 was significantly decreased,compared with the blank control (P＜0.01).Conclusions The leukoplakia chemokines CXCL9 and CXCL10 are highly expressed in vitiligo patients.The tacrolimus can significantly reduce the expression of CXCL9 and CXCL10 in keratinocytes under stress,and it therefore plays a therapeutic role in vitiligo.

LncRNA53106 regulates CXCL10 and affects the apoptosis of islet β cells / 中华内分泌代谢杂志

Objective To investigate the regulatory mechanism of long non-coding RNA ( lncRNA) 53106 in the apoptosis model of MIN6 cells stimulated by cytokines. Methods The stimulation model of cytokines 10 ng/ml interleukin-1β, 50 ng/ml tumor necrosis factor-α, 50 ng/ml interferon-γin MIN6 islet cell lines were established. The apoptosis rate was measured by flow cytometry and the expression levels of lncRNA53106 and ( C-X-C motif) ligand (CXCL)10 were detected by realtime quantitative PCR (qPCR). LncRNA53106 smart silencer and CXCL10 siRNA were constructed, and lncRNA53106 and CXCL10 were knocked down respectively. Then inflammatory cytokines were combined to stimulate, and their roles in the apoptosis of min6 cells were detected by flow cytometry, qPCR, and Western blotting. Results In the apoptosis model of MIN6 cells stimulated by cytokines, the apoptosis rate of cytokines group was significantly increased and reached statistical significance. The apoptosis rate of the knocked down lncRNA53106 group was significantly lower than that of the control group ( P<0.05) . The expression of CXCL10 was also decreased in the knockdown group by qPCR and Western blotting, the expressions of the apoptosis-related factors Bax and Caspase3 mRNA were decreased. The apoptosis rate in the knocked down CXCL10 group was significantly lower than that in the control group (P<0.05), and the expression of lncRNA53106 was slightly increased, but the difference was not significant ( P=0.61) . Conclusion LncRNA53106 may promote the expression of apoptosis factor by upregulation of CXCL10, and promote the apoptosis ofβcells of the pancreas, which may lead to the occurrence of type 1 diabetes.

Chemokine Receptor CXCR3 in the Spinal Cord Contributes to Chronic Itch in Mice / 神经科学通报·英文版

Recent studies have shown that the chemokine receptor CXCR3 and its ligand CXCL10 in the dorsal root ganglion mediate itch in experimental allergic contact dermatitis (ACD). CXCR3 in the spinal cord also contributes to the maintenance of neuropathic pain. However, whether spinal CXCR3 is involved in acute or chronic itch remains unclear. Here, we report that Cxcr3 mice showed normal scratching in acute itch models but reduced scratching in chronic itch models of dry skin and ACD. In contrast, both formalin-induced acute pain and complete Freund's adjuvant-induced chronic inflammatory pain were reduced in Cxcr3 mice. In addition, the expression of CXCR3 and CXCL10 was increased in the spinal cord in the dry skin model induced by acetone and diethyl ether followed by water (AEW). Intrathecal injection of a CXCR3 antagonist alleviated AEW-induced itch. Furthermore, touch-elicited itch (alloknesis) after compound 48/80 or AEW treatment was suppressed in Cxcr3 mice. Finally, AEW-induced astrocyte activation was inhibited in Cxcr3 mice. Taken together, these data suggest that spinal CXCR3 mediates chronic itch and alloknesis, and targeting CXCR3 may provide effective treatment for chronic pruritus.

Lipopolysaccharide up-regulates IP-10 expression through the activation of NF-κB in rat peritoneal mesothelial cells / 中国医师杂志

Objective To observe the expression of interferon induced protein (IP)-10 and the role of nuclear factor-kappaB (NF-κB) signaling pathway in rat peritoneal mesothelial cells (RPMCs) under the action of lipopolysaccharide (LPS).Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and cultured under defined in vitro conditions.The cells were exposed respectively to different concentrations of LPS (0,10,100,1 000,10 000 ng/ml) for 3 h or treated with LPS (100 ng/ml)for different time points (0,1,3,6,12,24,48 h).For observing the effect of LPS on the expression of p-p65 and p65,the RPMCs were treated with LPS (100 ng/ml) for different time points (0,15,30,60,120 min).For observing the effect of BAY11-7085 on the expression of IP-10 mRNA,the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μ mol/L) for2 h,then treated with LPS for another 3 h,respectively.Expression of IP-10 mRNA was examined by reverse transcription-polymerase chain reaction (RT-PCR).Expression of NF-κB and p-NF-κB protein was detected by Western blot.The secretion of IP-10 was determined by enzyme-linked immunosorbent assay (ELISA).Results Compared with the control group,stimulation of RPMCs with 10 ng/ml LPS resulted in a significant increase in the expression of IP-10 mRNA (P ＜0.05).1 000 ng/ml LPS has the strongest effect on IP-10 expression compared with that of 10 ng/ml and 100 ng/ml LPS.Treatment with 100 ng/ml LPS resuhed in time-dependent increase in the gene level of IP-10,with the peak at 3 h.However,after that time point,the gene level of them was gradually attenuated.Following treatment with LPS (100 ng/ml),the level of p-NF-κB began to increase at 15 min,gradually reached the peak at 1 hour,and then decreased.But the level of which at 2 h is still significant higher than that of medium control.5 μmol/L BAY11-7085 significantly decreased the up-regulation of IP-10 induced by LPS.Conclusions LPS enhanced the expression of IP-10 on RPMCs in a concentration-dependent and a time-dependent manner.LPS induced expression of IP-10 depended on the NF-κB signal transduction pathway.

Expression of CXCL10 ,CXCR3 and T cell subsets in peripheral blood and effect of compound Chinese medicine on it in patients with advanced vitiligo / 重庆医学

Objective To detect the expression levels of peripheral blood CXCL10 and its receptor CXCR3 and T cell subsets in of the patients with advanced vitiligo and the influence of compound Chinese medicine on it.Methods Flow cytometry was used to detect the cellular proportions of peripheral blood T cell subsets,ELISA was employed to quantify serum CXCL10 and CXCR3 expression levels before and after treatment.Results After 1 month of taking Chinese medicine,the proportions of CD3+ CD4+ cells and CD3+ CD8+ cells were increased compared before treatment(P＜0.05).The expression level of peripheral serum CXCL10 before treatment was significantly increased compare with the healthy control group(P＜0.01),and the CXCL10 level after treatment was decreased significantly compared with that before treatment(P＜0.05).The expression level of peripheral serum CXCR3 was significantly increased compared with the healthy control group(P＜0.05),while which after treatment was still significantly higher than that in the healthy control group(P＜0.05).Conclusion CXCL10,CXCR3 and T cell subsets proportion may be involved in the pathogenesis of vitiligo.The compound Chinese medicine used in this study plays the curative effect possibly by regulating T cell subsets and expression levels of CXCL10 and CXCR3.

Association of gene polymorphism of CXCL10 and CXCL12 with tuberculosis susceptibility / 天津医药

Objective To explore the relationship of chemokines CXCL10-135G/A and CXCL12 -801G/A gene polymorphisms with susceptibility to tuberculosis. Methods CXCL10-135G/A and CXCL12-801G/A polymorphisms of 102 tuberculosis patients(case group)and 115 healthy controls(control group)were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and the relationship between the two polymorphisms and susceptibility to tuberculosis were analyzed. Results The genotype analysis of CXCL10-135G/A and CXCL12-801G/A was in accord with the law of Hardy-Weinberg equilibrium in the case group and the control group. The differences of genotype and allele distribution frequency of CXCL10-135G/A were statistically significant between the case group and the control group(all P<0.05).The frequency of G allele distribution was higher in the case group than that in the control group, and the frequency of A allele distribution was lower than that in the control group.There were no significant differences in genotype and allele distribution frequency of CXCL12-801G/A polymorphism between the case group and the control group (all P>0.05).Conclusion Chemokine CXCL10-135G/A gene polymorphism is associated with susceptibility to pulmonary tuberculosis,and CXCL12-801G/A gene polymorphism may not be associated with tuberculosis infection.

Effects of tacrolimus on the secretion of chemokines CXCL9 and CXCL10 by γ-interferon-simulated HaCaT cells / 中华皮肤科杂志

Objective To analyze effects of tacrolimus on the secretion of chemokines CXCL9 and CXCL10 by γ-interferon (IFN-γ)-simulated HaCaT cells,as well as phosphorylated Janus kinase 1 (p-JAK1) and phosphorylated signal transducer and activator of transcription 1 (p-STAT1),and to explore the mechanism of tacrolimus in the treatment of vitiligo.Methods HaCaT cells were treated with l,10,20,40,60,80,100,120 mg/L tacrolimus solution separately for 4 hours,and methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the cellular proliferative activity.HaCaT cells were divided into 4 groups:blank control group receiving no treatment,IFN-γgroup treated with 500 U/ml IFN-γfor 12 or 48 hours,tacrolimus group treated with 20 mg/L tacrolimus for 4 hours,and tacrolimus + IFN-γgroup treated with 20 mg/L tacrolimus for 4 hours followed by the treatment with 500 U/ml IFN-γfor 12 or 48 hours.Real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expression of CXCL9 and CXCL10,Western blot analysis to determine the protein expression of CXCL9,CXCL10,p-JAK1,and p-STAT1,and enzyme-linked immunosorbent assay (ELISA) to detect the levels of CXCL9 and CXCL10 in the culture supernatants of HaCaT cells.Results Tacrolimus at the maximum concentration of 20 mg/L had no effect on the proliferation of HaCaT cells (P ＞ 0.05).After the pretreatment with 20 mg/L tacrolimus,the mRNA expression of CXCL9 and CXCL10 significantly decreased from 10 369.08 ± 7.99 and 290.02 ± 2.16 to 5 914.33 ± 4.59 and 114.96 ± 0.73,respectively,after the treatment with IFN-γ(both P ＜ 0.01),and the protein expression of CXCL9,CXCL10,p-JAK1,and p-STAT1 also significantly decreased from 8.47 ± 0.29,7.87 ± 0.17,4.20 ± 0.18 and 4.29 ± 0.11 to 7.36 ± 0.13,7.36 ± 0.09,2.60 ± 0.16 and 3.62 ± 0.19,respectively,after the treatment with IFN-γ (all P ＜ 0.01).Moreover,the levels of CXCL9 and CXCL10 in the culture supernatants of HaCaT cells significantly decreased in the IFN-γgroup (1 213.36 ± 0.95,1 722.41 ± 2.57,respectively) compared with the tacrolimus + IFN-γ group (426.45 ± 0.31,554.12 ± 0.56,respectively,both P ＜ 0.01).Conclusion Tacrolimus can inhibit the secretion of CXCL9,CXCL10,p-JAK1 and p-STAT1 by HaCaT cells stimulated by IFN-γ.

Analysis of the immunological biomarker profile during acute Zika virus infection reveals the overexpression of CXCL10, a chemokine linked to neuronal damage

BACKGROUND Infection with Zika virus (ZIKV) manifests in a broad spectrum of disease ranging from mild illness to severe neurological complications and little is known about Zika immunopathogenesis. OBJECTIVES To define the immunologic biomarkers that correlate with acute ZIKV infection. METHODS We characterized the levels of circulating cytokines, chemokines, and growth factors in 54 infected patients of both genders at five different time points after symptom onset using microbeads multiplex immunoassay; comparison to 100 age-matched controls was performed for statistical analysis and data mining. FINDINGS ZIKV-infected patients present a striking systemic inflammatory response with high levels of pro-inflammatory mediators. Despite the strong inflammatory pattern, IL-1Ra and IL-4 are also induced during the acute infection. Interestingly, the inflammatory cytokines IL-1β, IL-13, IL-17, TNF-α, and IFN-γ; chemokines CXCL8, CCL2, CCL5; and the growth factor G-CSF, displayed a bimodal distribution accompanying viremia. While this is the first manuscript to document bimodal distributions of viremia in ZIKV infection, this has been documented in other viral infections, with a primary viremia peak during mild systemic disease and a secondary peak associated with distribution of the virus to organs and tissues. MAIN CONCLUSIONS Biomarker network analysis demonstrated distinct dynamics in concurrence with the bimodal viremia profiles at different time points during ZIKV infection. Such a robust cytokine and chemokine response has been associated with blood-brain barrier permeability and neuroinvasiveness in other flaviviral infections. High-dimensional data analysis further identified CXCL10, a chemokine involved in foetal neuron apoptosis and Guillain-Barré syndrome, as the most promising biomarker of acute ZIKV infection for potential clinical application.

Protection mediated by chemokine CXCL10 in BALB/c mice infected by Leishmania infantum

BACKGROUND Visceral leishmaniasis (VL) caused by Leishmania infantum is characterised by the loss of the ability of the host to generate an effective immune response. Chemokines have a direct involvement in the pathogenesis of leishmaniasis, causing a rapid change in the expression of these molecules during infection by Leishmania. OBJECTIVES Herein, it was investigated the role of CXCL10 in controlling infection by L. infantum. METHODS RAW 264.7 macrophages were infected with L. infantum in vitro and treated or not with CXCL10 (25, 50 and 100 ng/mL). Parasite load, as well as nitric oxide (NO), IL-4 and IL-10 production were assessed at 24 and 48 h after infection. In vivo, BALB/c mice were infected and treated or not with CXCL10 (5 μg/kg) at one, three and seven days of infection. Parasite load, IFN-g, IL-4, TGF-β and IL-10 were evaluated one, seven and 23 days post treatment. FINDINGS In vitro, CXCL10 reduced parasitic load, not dependent on NO, and inhibited IL-10 and IL-4 secretion. In vivo, CXCL10 was able to reduce the parasite load in both liver and spleen, four weeks after infection, representing a higher decrease in the number of parasites in these organs, also induced IFN-γ at day 23 after treatment, correlating with the decrease in parasite load, and reduced IL-10 and TGF-β. MAIN CONCLUSIONS This study suggests a partial protective role of CXCL10 against L. infantum, mediated by IFN-g, not dependent on NO, and with suppression of IL-10 and TGF-β. These data may provide information for the development of new approaches for future therapeutic interventions for VL.

Liver-targeted expression of chemokine CXCL10 inhibits ConA-induced liver injury / 中国免疫学杂志

Objective:To explore the effect and mechanism of CXCL10 on immune-mediated liver injury.Methods:The CXCL10 expression vector was injected into mice by hydrodynamic injection.The levels of CXCL10 in the liver were measured 72 hours after injection.Concanavalin A (ConA) was injected into mice to induce immune-mediated liver injury.The levels of alanine a minotransferase (ALT),tissue damage,IFN-γ,TNF-α and percentages and numbers of regulatory T cells were tested and compared between CXCL10 expression group and control group.Results:The levels of CXCL10 in the liver were significantly increased at 72 hours after CXCL10 expression vector was injected.The pretreatment of CXCL10 expression vector markedly reduced ConA-induced ALT levels,IFN-γlevels and TNF-α levels.Additionally,the pretreatment of CXCL10 expression vector increased the percentages and numbers of regulatory T ceils in the liver.Conclusion:The pretreatment of CXCL10 expression vector decreases the levels of inflammatory cytokines and attenuates ConA-induced liver injury,which might be beneficial for the treatment for immune-mediated liver injury.

Regulation of salvianolic acid A,C molecular pair-drug compatibility on CCL2 and CXCL10 in HK-2 cells and antioxidation / 重庆医学

Objective To study the regulation effect of salvianolic acid A and C molecular compatibility on inflammatory chemokine monocyte chemotaxis protein 1(CCL2)and CXC chemokine ligand 10(CXCL10)in human renal tubular epithelial(HK-2)cells intervened by human serum albumin(HSA),and to explore the anti-inflammatory and antioxidation effects of salvianolic acid A,C molecular medicine-pair compatibility.Methods The cultured human renal tubular epithelial cells were randomly divided into 5 groups:control group,model group,salvianolic acid A group(20 μmol/L),salvianolic acid C group(20 μmol/L),salvianolic acid A+C group(10 μmol/L salvianolic acid A+10 μmol/L salvianolic acid C).Except for the control group,the other groups were added with HSA intervention for 24,48,72 h,then the each treatment group was simultaneously added with the drug treatment.The enzyme linked immunosorbent assay(ELISA)method was used to detect the expression of CCL2,CC chemokine 7(CCL7)and CXCL10,and SOD,GSH and MDA were detected by WST-1,DTNB,TBA.Results Compared with the control group,the levels of CCL2,CCL7,CXCL10 and MDA at 3 time points of 24,48 72 h in other groups were significantly increased,while the GSH and SOD levels were significantly decreased(P＜0.05).Compared with the model group,the levels of CCL2,CCL7,CXCL10 and MDA in each treatment group were relatively decreased,while the GSH and SOD levels were relatively increased(P＜0.05).In the comparison of various treatment groups,the levels of CCL2,CCL7,CXCL10 and MDA in the salvianolic acid A+C group were lower than those in the salvianolic acid A group and salvianolic acid C group(P＜0.05),while the GSH and SOD levels were higher than those in the salvianolic acid A group and salvianolic acid C group(P＜0.05).Conclusion The salvianolic acid A and B molecular medicine-pair compatibility may improve renal fibrosis by decreasing the expression of inflammatory chemokines and antioxidation.