Anti-PD-L1 antibody enhances curative effect of cryoablation via antibody-dependent cell-mediated cytotoxicity mediating PD-L1highCD11b+ cells elimination in hepatocellular carcinoma

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1highCD11b+ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8+ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1highCD11b+ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1highCD11b+ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.

The CXCL9/10/11-CXCR3 axis as a predictor of COVID-19 progression: a prospective, case-control study

ABSTRACT Background: This study examined the relationship between levels of the chemokines CXCL9, CXCL10, CXCL11, and CXCR3 and mortality in patients with COVID-19.. Methods: A total of 71 patients hospitalized with COVID-19 and 35 health workers with no symptoms and negative SARS-CoV-2 PCR results were included in the study. CXCL9, CXCL10, CXCL11, and CXCR3 levels were measured in blood samples using enzyme-linked immunosorbent assays. Participants were divided into three groups: healthy individuals, patients with mild to moderate pneumonia, and patients with severe pneumonia. Patients were also divided into sub-groups according to the outcome: dead and survived. Results: Serum CXCL9, CXCL10, CXCL11, and CXCR3 levels were significantly higher in patients with severe COVID-19 than in those with non-severe COVID-19; were higher in both patient groups than in the control group; and were higher in patients who died than in those who survived. Lymphocyte counts, and fibrinogen and PaO2/FiO2 levels were significantly lower in patients with severe COVID-19 than in those with moderate disease. Patients with COVID-19 also had elevated neutrophil/lymphocyte ratios, neutrophil counts, and lactate dehydrogenase, C-reactive protein, D-dimer, and ferritin levels. Conclusions: This study confirmed that CXCL9, CXCL10, CXCL11, and CXCR3 levels are associated with disease severity in patients with COVID-19. These laboratory parameters can help to estimate disease severity and predict outcomes, and are useful in clinical decision-making.

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.

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.

CXCL9 mRNA expression in ovarian tumor tissue and its relations with prognosis and characteristics of immune microenvironment / 上海交通大学学报（医学版）

Objective : To investigate the effects of CXCL9 (C-X-C motif chemokine ligand 9) mRNA expression on the overall survival of ovarian cancer patients, and to explore its relations with immune-related pathways and gene expression in tumor microenvironment, so as to re-veal the significance of CXCL9 in the prognosis of ovarian cancer. Methods ¡¤ Kaplan-Meier method was used to analyze the relationship be-tween CXCL9 mRNA expression and the survival of ovarian cancer patients in The Cancer Genome Atlas (TCGA) database. Gene Set Enrich-ment Analysis (GSEA) was used to assess the biological function of CXCL9 mRNA in ovarian cancer. The correlation of CXCL9 mRNA with cluster of differentiation 8A (CD8A) and immune checkpoint mRNA expression was analyzed. Results ¡¤ The high expression of CXCL9 mRNA was significantly associated with better prognosis in patients with ovarian cancer. GSEA showed that CXCL9 mRNA was enriched in the immune response-related pathway. In addition, Pearson correlation analysis showed that CXCL9 mRNA expression was positively correlated with the mRNA expression of CD8A and immune checkpoint. Conclusion ¡¤ The high expression of CXCL9 mRNA in ovarian tumor tissue is a good predictor of prognosis, and the mRNA expression of CXCL9 may be closely related to the recruitment of lymphocytes from tumor margin to the tumor microenvironment to exert the function of anti-tumor immune response.

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.

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.

Clinical significance of CXCL9 expression in peripheral blood of patients with systemic lupus erythematosus / 重庆医学

Objective To investigate the expression and significance of CXCL 9 in systemic lupus erythematosus (SLE ) . Methods The level of CXCL9 in peripheral blood from 20 patients with SLE and 20 normal controls were measured by enzyme linked immunosorbent assay .Compare the difference of the expression level of CXCL 9 in peripheral blood between two groups and analyzed their correlation with ages ,duration ,SLE diseases activity index (SLEDAI) .Results The level of CXCL9 in peripheral blood in patients with systemic lupus erythematosus was (1 549 .14 ± 362 .74)pg/L ,but in normal controls was(602 .54 ± 83 .70)pg/L .The level of CXCL9 in peripheral blood in patients with systemic lupus erythematosus was obviously higher than that in controls by statistical analysis ,there was significant difference between the two groups (P<0 .01) .The expression level of CXCL9 in pe‐ripheral blood of patients with systemic lupus erythematosus was positively correlated with SLEDAI (r=0 .892 ,P<0 .01) .Conclu‐sion CXCL9 may be involved in the pathogenesis of systemic lupus erythematosus .

Effects of Danzhi Jiangtang Capsule on CXCL9 and IL-8 in Diabetic Model Rats / 中国中医药信息杂志

Objective To investigate the effects of Danzhi Jiangtang Capsule on CXCL9 and IL-8 in type 2 diabetic model rats, and discuss the mechanism in TCM intervention in type 2 diabetes. Methods Fifty male SD rats were randomly divided into 5 groups:normal control group, high-dosage group of Danzhi Jiangtang Capsule, low-dosage group of Danzhi Jiangtang Capsule, pioglitazone group and model group. Normal control group was fed with basal diet, while the other groups were fed with high fat diet. A single intraperitoneal injection of STZ of 35 mg/kg was employed to establish the animal model of diabetes mellitus. When the diabetic rodent model was built, high-dosage group of Danzhi Jiangtang Capsule and low-dosage group of Danzhi Jiangtang Capsule were treated with 1.08 g/(kg?d) and 0.54 g/(kg?d) by gavage, pioglitazone group was treated with 10 mg/(kg?d) by gavage, the blank group and the model group were treated with physiological saline 5 mL/(kg?d) by gavage for 8 weeks. CXCL9, IL-8 and blood glucose, and lipid in serum of rats were determined with double antibody sandwich method. Results Expressions of CXCL9 and IL-8 in model group increased significantly, compared with the normal control group (P<0.05, P<0.01). Expressions of CXCL9 and IL-8 in high and low dosage Danzhi Jiangtang Capsule group significantly decreased, the blood glucose and lipid also decreased, compared with model group (P<0.01). Conclusion Danzhi Jiangtang Capsule has inhibitory effects on the high expression of inflammatory factors in diabetes, and can also lower bloodglucose, and regulate blood lipid.

Evaluating the potential value of MIG and IFN-γ as biomarkers for the differential diagnosis of active pulmonary tuberculosis / 中国医师杂志

Objective To study the feature of MIG and IFN-γ obtained from PBMCs stimulated with Mtb specific antigens and the potential value in the differential diagnosis of active pulmonary tuberculosis from bacterial pneumonia and primary lung cancer. Methods 90 patients with active pulmonary tuberculosis and 31 patients with bacterial pneumonia and primary lung cancer were enrolled. MIG and IFN-γin supernatants from PBMCs stimulated with Mycobacterium tuberculosis-specific antigens were analyzed with Bender Flowcytomix on flow cytometry. The diagnostic values were established based on receiver operating characteristic curve analysis. Results PBMCs stimulated with Mtb-specific antigens produced significantly higher levels of MIG compared with IFN-γ The level of MIG in active pulmonary TB patients was significantly higher than in controls(3023.0 pg/ml vs 112.5 pg/ml, P ＜0.0001). The MIG and IFN-γtests were positive in 96. 8 and 86. 7% of the TB patients, the specificity was up to 94. 4 and 87. 1%. With combination of MIG and IFN-γtests, the positive rate increased among TB patients to 97. 8% without a significant decrease in specificity. Conclusions The responses of the MIG and IFN-γagainst to Mtb-specific antigens could be used to discriminate newly-treated active pulmonary tuberculosis fiom bacterial pneumonia and primary lung cancer. Combination of MIG and IFN-γ might be a simple and quick approach to diagnosis newly-treated active pulmonary tuberculosis.

Expression of Th1/Th2 chemokines and their receptors in the lesions of bullous pemphigoid / 中国免疫学杂志

Objective:To investigate the expression of Th1 chemokine CXCL9,CXCL10,CXCL11,Th2 chemokine CCL22 and their receptors in the lesions of bullous pemphigoid (BP).Methods:Immunohistochemical assay was performed to detect the expression of CXCL9,CXCL10,CXCL11,CCL22 and their receptors CXCR3 and CCR4 in BP lesions and normal control skin.Results:CXCL9,CXCL10,CXCL11,CCL22,CXCR3 and CCR4 were overexpressed in BP lesions than those in normal control skin (P＜0.01).The positive rates of CXCL9,CXCL10,CXCL11 and CXCR3 in BP lesions were 50%(15/30),46.7%(14/30),46.7%(14/30) and 53.3%(16/30),respectively.The positive rates of CCL22 and CCR4 were 66.7% (20/30) and 56.7% (17/30).Conclusion:The overexpression of Th1 chemokine CXCL9,CXCL10,CXCL11,Th2 chemokine CCL22 and their receptors may play important roles in the pathogenesis of BP.

Progress in the Study of Chemokine CXCL9/Mig / 中国生物工程杂志

Chemokine CXCL9/Mig (monokine induced by IFN-?) belongs to the subfamily of chemotactic cytokines known as CXC-chemokines. In vivo CXCL9 is mainly induced by IFN-? in macrophages and primary glial cells. In vitro, CXCL9 can be secreted by cells such as macrophages, microvascular endothelial cells and neutrophils, in response to the synergy of IFN-? and TLR(toll-like receptor) ligands. CXCL9 is a chemoattractant for activated T lymphocytes, tumor-infiltrating T-lymphocytes, but not for neutrophils or monocytes. The receptor specific for CXCL9 is CXCR3, a G protein-coupled protein which has seven transmembrane domain. The structure and the chemical characterization of CXCL9, as well as its effects on autoimmune deseases, allograft rejection, cancer therapy were reviewed.