Relationship between CCL21 and TREM2/DAP12 signaling pathways in spinal dorsal horn in remifentanil-induced hyperalgesia in mice with incisional pain / 中华麻醉学杂志

Objective:To evaluate the relationship between CCL21 and triggering receptor expressed on myeloid cells 2 (TREM2)/DNAX-activating protein of 12 kDa (DAP12) signaling pathways in the spinal dorsal horn in remifentanil-induced hyperalgesia in mice with incisional pain.Methods:Thirty-two SPF healthy male C57BL/6J mice, weighing 18-22 g, aged 8-10 weeks, were divided into 4 groups ( n=8 each) using a random number table method: control group (group C), CCL21 neutralizing antibody group (group anti-CCL21), remifentanil + incisional pain group (group R+ I), and CCL21 neutralizing antibody + remifentanil + incisional pain group (group anti-CCL21+ R+ I).A CCL21 neutralizing antibody 0.3 μg (diluted to 10 μl in normal saline) was intrathecally injected in anti-CCL21 and anti-CCL21+ R+ I groups twice a day.Normal saline 10 μl was intrathecally injected at the same time point twice a day in C and R+ I groups.Fifteen min after intrathecal injection, normal saline 0.1 ml was injected via the caudal vein for 4 consecutive times at an interval of 15 min in C and anti-CCL21 groups, and remifentanil 10 μg/kg (diluted to 0.1 ml in normal saline) was injected via the caudal vein for 4 consecutive times at an interval of 15 min in R+ I and anti-CCL21+ R+ I groups.The tail-flick latency (TFL) and mechanical paw withdrawal threshold (MWT) were measured at 24 h before remifentanil or normal saline injection (T 0) and 3, 6, 24 and 48 h after stopping injection of remifentanil or normal saline (T 1-4).The mice were sacrificed after the last measurement of pain threshold, and L 4-6 segments of the spinal cord were removed for determination of the expression of TREM2 and DAP12 protein and mRNA (by Western blot or quantitative real-time polymerase chain reaction). Results:Compared with group C, TFL was significantly shortened and MWT was decreased at T 1-4, and the expression of TREM2 and DAP12 protein and mRNA was up-regulated in group R+ I and R+ I+ anti-CCL21 ( P<0.05), and no significant change was found in the parameters mentioned above in group anti-CCL21 ( P>0.05).Compared with group R+ I, TFL was significantly prolonged and MWT was increased at T 1-4, and the expression of TREM2 and DAP12 protein and mRNA was down-regulated in group anti-CCL21+ R+ I ( P<0.05). Conclusions:CCL21 is involved in remifentanil-induced hyperalgesia by activating TREM2/DAP12 signaling pathways in the spinal dorsal horn of mice with incisional pain.

Association of serum chemokine ligand 21 levels with asthma control in adults

OBJECTIVES: The chemokine ligand (CCL) 21 regulates the maturation, migration, and function of dendritic cells, and has been implicated in the pathogenesis of asthma. This study aimed to investigate the association between serum CCL21 levels and asthma control. METHODS: The serum levels of CCL21 and other inflammatory cytokines were analyzed in patients with asthma (n=44) and healthy controls (n=35) by enzyme-linked immunosorbent assay. IgE levels and eosinophil counts were determined by turbidimetric inhibition immunoassay and fully automatic blood analysis, respectively. The Asthma Control Test (ACT) questionnaire was used, and spirometry and fractional exhaled nitric oxide (FENO) measurements were performed. A multiple unpaired Student's t-test was performed to analyze the differences in CCL21 and interleukin levels between patients with asthma and healthy controls. The correlation of CCL21 levels with disease severity was evaluated using the Pearson's rank correlation test. RESULTS: Serum CCL21 levels were lower in patients with asthma (254.78±95.66 pg/mL) than in healthy controls (382.95±87.77 pg/mL) (p<0.001). Patients with asthma had significantly higher levels of IL-1β (19.74±16.77 vs. 2.63±5.22 pg/mL), IL-6 (7.55±8.65 vs. 2.37±2.47 pg/mL), and tumor necrosis factor-α (12.70±12.03 vs. 4.82±3.97 pg/mL) compared with the controls. CCL21 levels were positively correlated with the ACT score (rs=0.1653, p=0.0062), forced expiratory volume in 1s (FEV1)/forced vital capacity (rs=0.3607, p<0.0001), and FEV1 (rs=0.2753, p=0.0003), and negatively correlated with FENO (rs=0.1060, p=0.0310). CCL21 levels were negatively correlated with serum IgE levels (rs=0.1114, p=0.0268) and eosinophil counts (rs=0.3476, p<0.0001). CONCLUSIONS: Serum CCL21 levels may be a new biomarker for assessing asthma control.

Chemokine CCL21 in anterior cingulate cortex is involved in chronic neuropathic pain in a rat model / 中国组织工程研究

BACKGROUND: The pathogenesis of chronic pathological pain is yet unknown. Some studies have shown that after spinal cord injury, CCL21 can activate microglia in the central nervous system and is expressed only in damaged neurons, promoting the formation of chronic pathological pain. OBJECTIVE: To investigate whether the anterior cingulate cortex is involved in the formation of chronic pathological pain after inferior orbital nerve ligation in rats, and whether blocking chemokine CCL21 in the anterior cingulate cortex can reduce the chronic neuropathic pain. METHODS: A total of 80 male Sprague-Dawley rats were randomly divided into 4 groups with 20 rats in each group. In the sham group, only the infraorbital nerve of the rats was exposed; in the model group, the left infraorbital nerve was ligated; in the anti-CCL21 group, CCL21 neutralizing antibodies was administered to the anterior cingulate cortex of the rats on the 7th day after surgery; and in the PBS control group, PBS solution was given into the anterior cingulate cortex of rats on the 7th day after surgery. Rats in the sham and model groups were subjected to behavioral tests on the 3rd, 5th, 7th, and 14th days after surgery, and those in the anti-CCL21 and PBS control groups were subjected to the behavioral test at 6 hours after administration. All rats were sacrificed under anesthesia after behavioral tests. The cortical tissues were taken from the anterior cingulate, and the protein content of CCL21 was determined by western blot and immunofluorescence. RESULTS AND CONCLUSION: The pain threshold of the rats in the model group was lower than that in the sham group, and the expression of CCL21 in the anterior cingulate cortex was significantly higher in the model group than the sham group. After the administration of CCL21 neutralizing antibody, the expression of CCL21 was reduced to some extents, and the rat pain threshold was increased accordingly. These findings reveal that the anterior cingulate cortex of rats may be involved in the production of chronic pathological pain, and the administration of CCL21 neutralizing antibody can relief the pain.

Progress in targeting therapy of cancer metastasis by CCL21/CCR7 axis / 生物工程学报

Metastasis is the leading cause of mortality for cancer patients, and lymphatic metastasis is one of the main ways of tumor metastasis. The role of CCL21 and its receptor CCR7 in lymphatic metastasis has been increasingly concerned in recent years. CCR7 is mainly expressed by both dendritic cells and T cells for immune responses. CCL21, the chemokine ligand for CCR7, secreted from lymphatic endothelial cells binds CCR7 and recruits immune cells toward lymphatic vessels and lymphatic nodes. CCR7 expressed tumor cells can also metastasize to lymphatic system by the similar way as immune cells. Targeting CCL21/CCR7 axis to inhibit lymphatic metastasis but remain potent anti-tumor immune response has increasingly become a spot light of tumor immunotherapy. In this review, we summarize the role of CCL21/CCR7 axis in lymphatic metastasis, as well as preclinical trials and clinical trials in targeting CCL21/CCR7 axis for tumor metastasis therapy, hoping to accelerate the progress on tumor metastasis therapy by targeting CCL21/CCR7 axis.

GLGZD improves cerebral ischemia-reperfusion injury in rats by inhibiting microglial activation / 中国药理学通报

Aim To study whether GLGZD exerts brain protection by affecting the activation of cortical microglia in cerebral ischemia-reperfusion rats. Methods The nylon thread plug was used to establish the MCAO model. After GLGZD treatment for seven days, mNSS was used to evaluate the neurological function of each group of rats, MRI to detect cerebral infarction volume in rats, TUNEL to detect the apoptotic rate of nerve cells, immunohistochemistry to detect TNF-α protein expression in ischemic cortical brain tissues, and RT-qPCR to detect mRNA expression of neuron-microglia interaction-related factors TWEAK, Fnl4, NIK, Rel B, CCL21, CXCR3 and microglial activation marker IBA-1 in ischemic cortical brain tissues. Results GL-GZD could significantly improve the neurological function of MCAO rats, and markedly reduce the infarct volume and apoptosis of ischemic cortical neurons in MCAO rats. It also could significantly down-regulate the expressions of TNF-a protein and TWEAK, Fnl4, NIK, Rel B, CCL21, CXCR3 and IBA-1 mRNA in ischemic cortex of MCAO rats. Conclusions GLGZD can significantly improve cerebral ischemia-reperfusion injury in rats, which may be related to inhibition of microglial cell activation by affecting TWEAK/Fn14/ CCL21/CXCR3 signaling pathway.

Relationships between serum expression levels of CCL21 and CCL19 in rheumatoid arthritis and interstitial lung disease / 中国免疫学杂志

Objective:To investigate the relationships between the expression levels of chemokines CCL19 and CCL21 in serum of patients with RA and interstitial lung disease,as well as the correlation between the chemokines and the clinical parameters.Methods:Patients hospitalized in the First Hospital of China Medical University from May 2016 to March 2017 were enrolled.Serum levels of CCL21 and CCL19 in the 62 patients with RA,among them,34 cases without pulmonary interstitial lesions while 28 cases with lung interstitial diseases,18 cases ⅡP and 20 health control were detected with enzyme-linked immunosorbent assay (ELISA).One-way ANOVA,LSD-t test,Kruskal-Wallis test,Pearson and Spearman correlation analysis were used for statistical analysis.Results:The levels of CCL21 and CCL19 were both higher in the RA patients than the health controls(P＜0.05;P＜0.01),The concentrations of CCL21 in serum of the RA patients with ILD were higher than thoes without ILD,higher than the health controls (P＜0.05;P＜0.01),serum levels of CCL21 in the ⅡP patients were higher than the health controls (P＜0.01),too.The serum levels of CCL21 in men were higher than women(P＜0.05).Serum CCL21 levels were negatively correlated with albumin and BMI(r=-0.280,P＜0.05;r =-0.605,P＜0.05).Levels of CCL19 were negatively correlated with the levels of C4 (r =-0.326,P＜0.05),and positively correlated with the levels of D-Dimer(r =0.592,P＜0.05).Conclusion:Chemokines CCL21 and CCL19 are highly expressed in serum of patients with RA,they may be involved in the pathogenesis of RA.Furthermore,CCL21 may has a correlation with the lung interstitial lesion in RA.

Clinical prediction value of chemokine CCL21 level for acute coronary syndrome / 中华老年医学杂志

Objective To investigate the clinical prediction value of chemokine CCL21 level for acute coronary syndrome.Methods Totally 212 patients receiving coronary arteriography were divided into acute myocardial infarction group(AMI,n=72),unstable angina pectoris group(UAP,n=76),and stable angina pectoris group(SAP,n=64).The serum level of chemokine CCL21 was detected by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA)and the pathological changes of coronary artery were measured by Gensini scoring system.All patients were followed up for six months and the cardiovascular adverse events were recorded.Results The serum level of CCL21 was(169.72±14.64)ng/L in AMI group,(154.42±16.50)ng/L in UAP group,and(143.87±9.80) ng/L in SAP group,with statistically significant differences (F =99.818,P =0.000).Serum levels of CCL21 in ACS group and SAP group were positively correlated with Gensini score(r=0.474,P =0.000;r=0.350,P=0.049).Binary Logistic regression analysis revealed that chemokine CCL21 was an independent risk factor for predicting acute coronary syndrome (OR =1.049,P =0.022).The CCL21-judged area under the ROC curve in acute coronary syndrome group was 0.887 ± 0.028 (P =0.000),with diagnostic point of serum level of chemokine CCL21 at 159.15 ng/L,sensitivity of 0.635,specificity of 0.981.Serum level of CCL21 was higher in the patients with cardiovascular adverse events than in the patients without cardiovascular adverse events[(168.57±7.24)ng/L vs.(156.92± 6.53) ng/L],with statistically significant difference (t =16.100,P =0.000).Conclusions Serum level of chemokine CCL21 reflects the severity degree of coronary artery disease.The chemokine CCL21,as an independent and effective marker,can predict acute coronary syndrome.

Level diversifies of plasma CCL19 and CCL21 in male drug users infected HCV / 中国免疫学杂志

Objective:To analyze the level diversifies of plasma CCL19 and CCL21 in the male drug users infected HCV.Methods: The plasma CCL19 and CCL21,anti-HCV and HCV RNA were detected by ELISA quantitation,ELISA qualitation and Real-time RT-PCR respectively.Compared with 60 healthy man,the level diversifies of plasma CCL19 and CCL21 in 391 male drug users conducted as part of HCV Surveillance Programme in Zhuhai were analyzed.Results: 180 of 391 male drug users were infected HCV and the infection rate was 46.04%.The level of plasma CCL19 and CCL21 in the male drug users[anti-HCV(-)/HCV RNA(-)] were higher than that in the healthy man (P≤0.001).The level of plasma CCL19 and CCL21 in anti-HCV(-)/HCV RNA(+) group was lower than that in the others(P≤0.05) .Compared with that in the healthy man,the level of plasma CCL19 and CCL21 in the drug abuse anti-HCV(+)/HCV RNA(-) group had significant deviation(P<0.05).Conclusion: Drug abuse can heighten the level of plasma CCL19 and CCL21 in man.Increasing the level of plasma CCL19 and CCL21 may conduce to spontaneous HCV clearance.It may prognosticate that HCV infection will be persistent and have a bad consequence when the level of plasma CCL19 and CCL21 were degraded.

Role of cemokine ligand 21 in spinal cord in tibia bone cancer pain in rats / 中华麻醉学杂志

Objective To investigate the role of chemokine ligand 21 (CCL21) in the spinal cord in tibia bone cancer pain (BCP) in rats.Methods Forty adult female SD rats weighing 160-180 g were randomly divided into 5 groups (n=8 each):sham operation group (group Ⅰ ); sham operation + CCL21 neutralizing antibody group (groupⅡ); BCP group (group [); BCP + PBS group (group Ⅳ); BCP + control IgG group (groupⅤ)and BCP + CCL21 neutralizing antibody group (group Ⅵ).BCP was induced by inoculating Walker-256 mammary gland carcinoma cells into the rat tibia medullary cavity in groups Ⅲ-Ⅵ.PBS 15 μl,IgG 10 μg and CCL21 neutralizing antibody 10 μg were injected intrathecally (IT) at 14 days after intra-tibial injection of Walker-256 mammary gland cancer cells in groups Ⅳ- Ⅵ respectively.Mechanical withdrawal threshold to yon Frey filament stimulation (MWT) was measured at 1 day before (To,baseline) ; 7 and 14 d after Walker-256 cell injection (T1,T2)and at 0.5,1,2,4,8,12,24 and 48 h after intrathecal injection (T3-10 ).Results Intra-tibial injection of Walker-256 mammary gland cancer cells significantly decreased MWT as compared with the baseline values in administration of CCL21 neutralizing antibody at T5-8 as compared with MWT before intrathecal administration at T2 in group Ⅵ.MWT was significantly lower in groups Ⅲ- Ⅳ than in groups Ⅰ and Ⅱ.MWT was significantly higher at T5-8 in group Ⅵ than in groups Ⅲ - Ⅴ.Conclu]sion CCL21 in the spinal cord is involved in the maintenance of tibia BCP in rats.

CCL21/CCR7 axis and mechanisms of tumor invasion and metastasis / 国际肿瘤学杂志

Secondary lymphoid tissue chemokine (CCL21) is a double-edged sword, which exerts antitumor, anti-infection immune response by binding to the receptor CCR7 on the surface of the multiple immune cells. However, a variety of tumor cells also express the receptor CCR7, the combination of CCL21 with CCR7promotes the invasion and metastasis of tumor cells, leading to the facilitation of tumor development. Therefore,exploring the mechanism(s) of tumor invasion and metastasis might be helpful for use of CCL21 as tumor gene therapy through blocking of CCL21's promotion of tumor invasion and metastasis.