Effect of siRNA interference with C-C chemokine ligand 5 on biological characteristics of laryngeal carcinoma cells / 中国生物制品学杂志

@#Objective To investigate the expression of C-C chemokine ligand 5(CCL5) in head and neck squamous cell carcinoma(HNSCC),and explore the effect of CCL5 on the biological characteristics of laryngeal carcinoma cells.Methods Gene Expression Profiling Interactive Analysis(GEPIA) database was used to investigate the expression of CCL5 in HNSCC.The laryngeal carcinoma cells TU177 were transfected with siRNA(siRNA group),and the control(NC) group was set up.The cell proliferation,migration,cycle and apoptosis of each group were detected by CCK8 assay,cell scratch test and flow cytometry respectively.RT-PCR and Western blot were used to detect the knock-down efficiency of CCL5 and the mRNA transcription and protein expression of multidrug resistance protein 2(MRP2) and bcl-2-associated x protein(Bax).Results The expression of CCL5 in HNSCC was higher than that in normal tissues(P <0.05).Compared with NC group,siRNA showed higher knock-down efficiency(t=12.898 and 22.656 respectively,each P <0.01);siRNA interference with CCL5 inhibited the proliferation and migration of laryngeal carcinoma cells,and promoted the late apoptosis of laryngeal carcinoma cells and the expression of apoptosis protein Bax(t=2.600～11.667,each P <0.05).Conclusion CCL5 was highly expressed in HNSCC,while siRNA interference with CCL5 inhibited the proliferation,migration and promoted apoptosis of laryngeal carcinoma cells TU177 by up-regulating the expression of Bax,which laid a foundation of the possibility of CCL5 as a new target for the treatment of laryngeal carcinoma.

Construction and functional analysis of EGFRvIII CAR-T cells co-expressing IL-15 and CCL19 / 生物工程学报

The aim of this study was to investigate the functional characteristics and in vitro specific killing effect of EGFRvIII CAR-T cells co-expressing interleukin-15 and chemokine CCL19, in order to optimize the multiple functions of CAR-T cells and improve the therapeutic effect of CAR-T cells targeting EGFRvIII on glioblastoma (GBM). The recombinant lentivirus plasmid was obtained by genetic engineering, transfected into 293T cells to obtain lentivirus and infected T cells to obtain the fourth generation CAR-T cells targeting EGFRvIII (EGFRvIII-IL-15-CCL19 CAR-T). The expression rate of CAR molecules, proliferation, chemotactic ability, in vitro specific killing ability and anti-apoptotic ability of the fourth and second generation CAR-T cells (EGFRvIII CAR-T) were detected by flow cytometry, cell counter, chemotaxis chamber and apoptosis kit. The results showed that compared with EGFRvIII CAR-T cells, EGFRvIII-IL-15-CCL19 CAR-T cells successfully secreted IL-15 and CCL19, and had stronger proliferation, chemotactic ability and anti-apoptosis ability in vitro (all P < 0.05), while there was no significant difference in killing ability in vitro. Therefore, CAR-T cells targeting EGFRvIII and secreting IL-15 and CCL19 are expected to improve the therapeutic effect of glioblastoma and provide an experimental basis for clinical trials.

Small molecule CCR5 antagonists in clinical trials and their drug resistances:research advances / 国际药学研究杂志

C-C chemokine receptor 5 (CCR5), one of the major co-receptors of HIV-1, can mediate the fusion of HIV-1 to cell membranes. CCR5 antagonists can bind to CCR5 and cause conformational changes in CCR5, thus blocking HIV-1 infection. Several small molecule CCR5 antagonists with strong activity and good tolerance have been screened and entered the clinical trials. With the widespread use of CCR5 antagonists, drug resistance has gradually emerged. There are many reports about of drug-related failure in vivo and in vitro. Therefore, this review summarizes the mechanism of CCR5-mediated HIV-1 infection, the research progress in maraviroc and other CCR5 antagonists which have entered clinical trials and their drug resistance.

Increased serum C-C chemokine ligand 19 levels correlated with B cell abnormalities in systemic lupus erythematosus / 北京大学学报(医学版)

Objective:To detect the levels of serum C-C chemokine ligand 19 (CCL19) in patients with systemic lupus erythematosus (SLE) and to evaluate the correlation between CCL19 expression and clinical features and laboratory parameters,trying to reveal the possible role of CCL19 in the pathogenesis of systemic lupus erythematosus.Methods:The levels of serum CCL19 were measured by enzyme linked immunosorbent assay (ELISA) in 90 patients with SLE and 30 healthy controls.These SLE patients included 75 patients who received treatment with glucocorticoids and disease-modifying anti-rheumatic drug (DMARD) and 15 patients without therapy.The frequencies of peripheral blood B cells and the B cell subsets were assessed in the patients with SLE by flow cytometry.The correlation between the clinical data,laboratory parameters,B cell subset frequencies and serum CCL19 levels were analyzed.Independent samples t test,paired t test,Pearson and Spearman correlation were used for statistical analyses.Results:The levels of CCL19 were markedly higher in the SLE patients without therapy and the patients with therapy than in the health controls[(596.25 ±409.19) ng/L and (422.90 ± 395.84) ng/L vs.157.79 ± 125.23) ng/L,all P ＜ 0.001].Serum CCL19 levels in the SLE patients without therapy were higher than the SLE patients who accepted glucocorticoids and DMARD treatment (P ＜ 0.05).The levels of serum CCL19 were positively correlated with anti-double stranded deoxyribonucleic acid (dsDNA),anti-nucleosome antibody (AnuA),IgA,IgG and IgM (r =0.38,P =0.007;r =0.332,P =0.029;r =0.519,P =0.007;r =0.461,P =0.018,respectively).Serum CCL19 levels in the SLE patients with photosensitivity,arthritis and secondary Sj(o)gren's syndrome were higher than the SLE patients without photosensitivity,arthritis and secondary Sj(o)gren's syndrome,respectively [(562.25 ± 399.12) ng/L,(565.6 ± 435.24) ng/L and (694.9 ± 531.02) ng/L vs.(394.7 ± 281.42) ng/L,(385.90-± 325.33) ng/L and (424.8 ± 305.46) ng/L,all P ＜ 0.05].The levels of serum CCL19 were positively correlated with the percentage of CD27-B cells and CD27-IgD-double-negative memory B cells (r =0.519,P =0.007;r =0.461,P =0.018,respectively).However,the levels of serum CCL19 were negatively correlated with the percentage of CD27 + memory B cells and CD27 + IgD-switched memory B cells (r =-0.433,P =0.027;r =-0.616,P =0.001,respectively).Conclusion:The increased serum CCL19 levels in SLE patients were associated with the production of autoantibodies,and CCL19 might be involved in the pathogenesis of SLE by disturbing the homeostasis of B cell subsets.

Effects of antagonistic peptides binding specifically with first and second extracellular loops of CCR5 on colitis rats induced by TNBS / 中国病理生理杂志

AIM:To study the effects of antagonistic peptides binding specifically with the first and second extracellular loops (ECL1 and ECL2) of C-C chemokine receptor 5 (CCR5) on the colitis rats induced by trinitrobenzenesulfonic acid (TNBS) and the mechanisms.METHODS:The colitis model of SD rats was induced by TNBS (100 mg/kg).The effects of 2 antagonistic peptides at different doses (ECL1:25, 35 and 45 mg/kg;ECL2:15, 25 and 35 mg/kg) on the model rats including the changes of disease activity index (DAI), colon macroscopic damage index (CMDI) and histological grading were observed.The mRNA and protein expression levels of TNF-α and COX-2 in the colonic mucosa were detected by real-time PCR and Western blot, respectively.RESULTS:Compared with model group, the changes of DAI, CMDI and histopathological injury of the rats treated with ECL2 antagonistic peptide HY at an appropriate dose were significantly reduced (P<0.05), and the protein and mRNA expression levels of TNF-α and COX-2 were significantly decreased (P<0.05).However, the effects of ECL1 antagonistic peptide GH on all scores and the expression levels of TNF-α and COX-2 were not obvious.CONCLUSION:ECL2 antagonistic peptide HY relieves TNBS-induced colitis in SD rats via down-regulating the expressions of TNF-α and COX-2 in the colonic mucosa, while the effect of ECL1 antagonist peptide GH was not obvious.

Correlation between CD4~+CD25~+T cells and CCR4 in human nasopharyngeal carcinoma / 西安交通大学学报(医学版)

Objective To explore the immune function of the whole body and the tumor site of human nasopharyngeal carcinoma (NPC) patients as well as its correlation with CCR4. Methods The ratios of CD4~+CD25~+T cells and CCR4+cells to lymphocytes were measured by flow cytometry in tissues (25 cases) and peripheral blood (35 cases) from nasopharyngeal carcinoma patients, and then statistical analysis was made. Results The ratios of CD4~+CD25~+T cells and CCR4+ cells in tissue and peripheral blood of NPC were higher than those in the control group (P<0.05). In NPC the ratios of the two cells in tissue were higher than in peripheral blood respectively (P<0.05), but there was no such difference between tissue and peripheral blood in the control group (P>0.05). The ratio of CD4~+CD25~+T cells in tissue at stage Ⅲ+Ⅳ was higher than that at stage Ⅰ+Ⅱ of NPC (P<0.05), but there was no such difference between the two stages in peripheral blood in NPC. There was a positive correlation between CD4~+CD25~+T cells and CCR4+ cells in tissue and peripheral blood of NPC (P<0.05). Conclusion NPC patients have different immunosuppression, and there is even more severe immunosuppression in the tumor site. The ratio of CD4~+CD25~+T cells is correlated with the stage of NPC. Therefore, as NPC progresses to later stages, the percentage of CD4~+CD25~+T cells is increased, which is correlated with poor prognosis. CCR4 plays an important role in reactant of CD4~+CD25~+T cells to tumor sites, and is resistant to immunosurveillance.

Expression of Mucosal Cyto-Chemokine mRNAs in Patients with Helicobacter pylori Infection

BACKGROUND: Helicobacter pylori-induced destruction of the gastroduodenal mucosal barrier is initiated with mucosal infiltration of inflammatory cells. Cytokines and chemokines have been suggested to play important roles in the migration and activation of these inflammatory cells into the mucosa. The present study aimed to investigate expression rates of cyto-chemokine mRNAs using gastric mucosal biopsy specimens. METHODS: In 98 patients infected with Helicobacter pylori, mucosal mRNA expression rates of cytokines (IL-1beta, IL-6, and IL-10), C-C chemokines (macrophage inflammatory protein 1alpha [MIP-1alpha], and macrophage inflammatory protein 1beta [MIP-1beta], monocyte chemotactic and activating factor [MCAF], regulated on activation, normal T cell expressed and presumably secreted [RANTES]) and C-X-C chemokines (IL-8 and growth regulated alpha [GRO-alpha]) were examined using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The expression rates of mRNA for IL-8, GRO-alpha, MIP-1alpha and RANTES were significantly more increased in H. pylori-positive patients than in H. pylori- negative patients. However, the expressions of IL-1beta, IL-6 and IL-10 mRNA were statistically not different between two groups. After eradication of H. pylori, expressions of mRNA for three cytokines (IL-1beta, IL-6 and IL-10), four C-C chemokines (MIP-1alpha, MIP-1beta, MCAF and RANTES) and two C-X-C chemokines (IL-8 and GRO-alpha) were significantly decreased. CONCLUSION: These results suggest that C-X-C chemokines and some C-C chemokines play important roles in H. pylori-associated peptic ulcer diseases.

C-C Chemokines Secretion of Lymphocyte and Monocyte in Chinese Neonate Cord Blood and Adult Peripheral Blood / 中国医师杂志

Objective To observe secretive ability of three C-C chemokines, macrophage inflammatory protein-1?(MIP-1?),MIP-1? and regulated-upon activation.normal T-cell expressed and secretory factor(RANTES)by lympocytes and moncytes of cord blood from chinese necnates and peripheral blood from chinese adults.Method Using Ficoll density-gradient centrifugation and gelatin-coated flask,the Iympocytes and monocytes were isolated from cord blood samples of seventeen Chinese neonates and peripheral blood samples of twenty Chinese adults.Purified lymphocytes and monocytes were incubated with PMA and LPS,respectively.Supernants were tested by ELISA for concentrations of MIP-1?,MIP-1? and RANTES.Results Concentrations of MIP-1?, MIP-1? and RANTES in cord blood lymphocyte and monocyte were (3920?730)pg/ml, (4910?590)pg/ml,(1470?410)pg/ml,(3240?980)pg/ml,(1960?1300)pg/ml and (240?120)pg/ml,respectively,and in adults peripheral blood were (6560?840)pg/ml,(5810?1150)pg/ml,(2250?570)pg/ml,(3010?1350) pg/ml,(2280?870)pg/ml,(690?430)pg/ml,respectively.Cord blood lymphocyte and monocyte showed diminished ability to secrete RANTES than adult peripheral blood.Conclusions The diminished RANTES secretion of cord blood lymphocyte and monocyte may be releted to the pathogenesis of Chinese neonatal HIV infection,and indicated some immunotherapies are feasible for preventing neonate from HIV mother-to-child transmission.