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Peri-implantitis is one of the leading causes of implant failure and loss, and its early diagnosis is not currently feasible due to the low sensitivity of currents methods. In the current exploratory cross-sectional study, we explored the diagnostic potential of lymphocyte B and Th17-chemotactic cytokine levels in peri-implant crevicular fluid (PICF) in 54 patients with healthy, peri-mucositis, or peri-implantitis implants. Peri-implant crevicular fluid was collected, and the levels of the molecules under study were quantified by Luminex assay. The concentrations of CCL-20 MIP-3 alpha, BAFF/BLYS, RANKL and OPG concentration in PICF were analyzed in the context of patient and clinical variables (smoking status, history of periodontitis, periodontal diagnosis, implant survival, suppuration, bleeding on probing, periodontal probing depth, clinical attachment level, mean of implant probing depth, and plaque index). Patients with peri-implantitis, appear to have an overregulation of the RANKL/BAFF-BLyS axis. This phenomenon needs to be investigated in depth in further studies with a larger sample size.
La periimplantitis es una de las principales causas de falla y pérdida del implante, y su diagnóstico temprano no es factible debido a la baja sensibilidad de los métodos actuales. En este estudio transversal exploratorio, se estudió el potencial diagnóstico de los niveles de citocinas quimiotácticas de linfocitos B y Th17 en el líquido crevicular periimplantario (LCPI) en 54 pacientes con implantes sanos, peri-mucositis o periimplantitis. Se recogió líquido crevicular periimplantario y se cuantificaron los niveles de las moléculas estudiadas mediante Luminex assay. Las concentraciones de CCL-20 MIP-3 alfa, BAFF/BLYS, RANKL y la concentración de OPG en LCPI se analizaron en el contexto de las variables clínicas y del paciente (tabaquismo, antecedentes de periodontitis, diagnóstico periodontal, supervivencia del implante, supuración, sangrado al sondaje, profundidad de sondeo periodontal, nivel de inserción clínica, media de la profundidad de sondeo del implante e índice de placa). Los pacientes con periimplantitis parecen tener una sobrerregulación del eje RANKL/BAFF-BLyS. Este fenómeno debe investigarse en profundidad en futuros estudios con un tamaño de muestra mayor.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Dental Implants/adverse effects , Peri-Implantitis/diagnosis , Biomarkers , Chile , Cross-Sectional Studies , Gingival Crevicular Fluid , Mucositis , RANK Ligand , Chemokine CCL20ABSTRACT
The role of chemokines/receptors in the pathophysiological activities such as inflammation and tissue injury has been clarified .In recent years, there have been increasing studies on their effect on tumors in the tumor microenvironment .This article reviews research advances in C -C motif chemokine ligand 20 (CCL20) and its receptor CCR6 in liver cancer and other tumors in recent years and analyzes the possible mechanism by which CCL20 and CCR6 promote tumor progression and their effect on tumor prognosis .Blocking CCL20/CCR6 interaction may provide a new direction for the targeted therapy for liver cancer.
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Objective To investigate the effects of narrow-band ultraviolet B (NB-UVB) therapy on the levels of plasmin and CC chemokine ligand 20 (CCL20) in peripheral blood of patients with psoriasis vulgaris.Methods A total of 60 patients with psoriasis vulgaris in progressive stage were treated with NB-UVB radiation thrice a week for 8 weeks.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of plasmin and CCL20 in the peripheral blood of the patients before and after the treatment,as well as in the peripheral blood of 50 healthy controls.Results After the treatment,psoriasis area and severity index (PASI) scores in patients were significantly decreased compared with those before the treatment (2.54 ± 1.64 vs.10.26 ± 3.14,t =17.40,P ＜ 0.05),and the response rate was up to 87％ (52/60).Before the treatment,levels of plasmin and CCL20 were both significantly higher in the patient group than in the control group (plasmin:180.07 ± 40.62 μg/L vs.76.30 ± 26.92 μg/L,t =15.45,P ＜ 0.05;CCL20:422.41 ± 129.87 pg/L vs.205.33 ± 49.89 pg/L,t =11.15,P ＜ 0.05).After the treatment,levels of plasmin (148.22 ± 40.05 μg/L) and CCL20 (329.67 ± 100.73 pg/L) in patients were significantly decreased compared with those before the treatment (t =4.97,6.44,P ＜ 0.05),but still significantly higher than those in controls (t =10.82,7.95,P ＜ 0.05).Before the treatment,the level of plasmin was positively correlated with the level of CCL20 in peripheral blood of the patients (r =0.57,P ＜ 0.05),and the levels of plasmin and CCL20 were both positively correlated with the PASI score (r =0.49,0.62,respectively,both P ＜ 0.05).Conclusion NB-UVB radiation may exert a therapeutic effect on psoriasis vulgaris by reducing levels of plasmin and CCL20 in peripheral blood of patients.
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Objective To evaluate effects of interleukin-36α (IL-36α) on psoriasiform skin lesions and C-C motif chemokine ligand 20 (CCL20) expression in mice.Methods Totally,30 BALB/c female mice were randomly and equally divided into 3 groups:control group treated with topical vaseline cream on the shaved back and intracutaneous injection with phosphate buffer saline (PBS),model group treated with topical imiquimod cream on the shaved back and intracutaneous injection with PBS,experimental group treated with topical imiquimod cream on the shaved back and intracutaneous injection with IL-36α solution.Psoriasis area severity index (PASI) was used to evaluate changes of psoriasiform skin lesions in mice,and light microscopy to observe morphological changes of skin lesions and to measure the thickness of the epidermis.Real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to determine the expression of IL-36α in skin lesions in the control group and model group,and qRT-PCR,Western blot analysis and immunohistochemical study to evaluate changes of CCL20 levels in skin lesions.Results The model group showed significantly increased mRNA (△ Ct value:0.0195 ± 0.0059) and protein expression (3.922 ± 0.248) of IL-36α compared with the control group (mRNA:0.0012 ± 0.0004,P ＜ 0.05;protein:0.690 ± 0.025,P ＜ 0.05).The mRNA and protein expression of CCL20 were significantly higher in the experimental group than those in the model group (mRNA:2.152 ± 0.793 vs.0.999 ± 0.178;protein:0.397 ± 0.033 vs.0.145 ± 0.030;both P ＜ 0.05),and higher in the model group than those in the control group (mRNA:0.378 ± 0.075;protein:0.025 ± 0.009;both P ＜ 0.05).Immunohistochemical study showed that the expression intensity of CCL20 in skin lesions significantly increased in the experimental group compared with that in the model group (Z =2.294,P ＜ 0.05).Conclusion IL-36α may aggravate psoriasiform skin inflammation in mice by promoting CCL20 expression.
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Objective:To construct recombinant adenovirus vector Ad5-CCL20,and detect the expression of CCL20 after Ad5-CCL20 transfected colon cancer cells CT-26.Methods:Genes encoding CCL20 was obtained from original plasmid double-digested with EcoR I/Sal I enzymes.The CCL20 DNA segments were linked into pDC316 to recombine shuttle plasmid pDC316-CCL20.After genome sequencing,we take shuttle plasmid pDC316-CCL20 and plasmid backbone pBHGIox_E1,3Cre co-transfecting 293T cells in mediation of liposome.The constructed recombinant adenovirus vector was named Ad5-CCL20.Lastly,after Ad5-CCL20 transfected CT-26 cells in vitro,the expression of CCL20 at different time points (12h,24h,36h and48h)was detected by Western blot and Elisa.Then,Culture supernatant was added into iDC and mDC to evaluate the chemotactic activity of CCL20.Results:The recombinant adenovirus Ad5-CCL20 were successfully constructed.The expression of CCL20 was detected by Western blot and Elisa.The level of CCL20 expression was increased with prolonged incubation of the infected CT-26 cells.Chemotaxis experiments show that the chemokine CCL20 had chemotactic activity to the iDC and mDC,but more obviouly for iDC (P＜0.05).Conclusion:The construction and obtain of recombinant adenovirus vector Ad5-CCL20 provide a new method for developing tumor immunotherapy.
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Objective To investigate the effects of rottlerin on in vitro proliferation of and expressions of interleukin (IL)-17C,CCL20 chemokine,and nuclear factor (NF)-κB in cultured human HaCaT keratinocytes.Methods Some HaCaT cells were divided into several test groups treated with rottlerin at concentrations of 0.5,1.0,2.0 and 4.0 μmol/L,a solvent group treated with RPMI 1640 culture solution containing the same volume of dimethyl sulfoxide (DMSO) as that of 4.0 μmol/L rottlerin,and a control group treated with RPMI 1640 culture solution.Cell counting kit-8 (CCK8) assay was conducted to estimate the proliferative activity of HaCaT cells after 24-,48-and 72-hour culture,RT-PCR to determine the mRNA expressions of IL-17C and CCL20 after 48-hour culture,and Western blot to measure the protein expressions of IL-17C,CCL20 and NF-κB after 48-hour culture.Statistical analysis was carried out by using repeated-measures analysis of variance,one-way analysis of variance and Pearson correlation analysis with the SPSS16.0 software.Results Rottlerin showed an inhibitory effect on the proliferation of HaCaT cells,and the inhibitory effect increased over time (F =126.936,P ＜ 0.05) and with the increase of rottlerin concentrations (F =838.308,P ＜ 0.05),with a significant interaction effect between rottlerin concentrations and treatment duration (F =15.961,P ＜ 0.05).After 48-hour treatment,a significant decrease was observed in the mRNA and protein expressions of IL-17C (F =206.041,233.887,respectively,both P ＜ 0.05) and CCL20 (F =143.883,162.431,respectively,both P ＜ 0.05),as well as in the protein expression of NF-κB (F =577.915,P ＜ 0.05) in the test groups with the increase in rottlerin concentrations.Conclusions Rottlerin can inhibit the proliferation of HaCaT cells in vitro,and decrease the mRNA and protein expressions of IL-17C and CCL20 likely by downregulating the protein expression of NF-κB.
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Objective To explore the expression and clinical significance of chemokine receptor 6 (CCR6)/chemokine ligand 20(CCL20) axis in hepatocellular carcinoma (HCC).Methods From March 2003 to December 2005,48 patients with HCC were selected,and one specimen of HCC tissue and one of corresponding adjacent tissue were taken from every patient.Another eight patients with benign liver lesions were selected,and one specimen of surgical sectioned normal liver tissue of each was taken.The relative expression quantity of CCR6 and CCL20 at mRNA level was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR).And the expression of CCR6 and CCL20 at protein level was determined by immunohistochemisty methods.One-way analysis of variance (ANOVA) was performed for comparison among groups of measurement data.Chi-square test was used for rate comparison.The correlation coefficient was calculated by Spearman's method.Survival curves were plotted by Kaplan-Meier method and the survival rate was compared by Log-rank test.Results The relative expression quantity of CCR6/CCL20 at mRNA level in HCC tissues (0.99±0.21 and 0.46± 0.11) were significantly higher than those of para carcinoma tissues (0.33 ± 0.09 and 0.31 ± 0.07) and normal liver tissues (0.22±0.06 and 0.28±0.05),and the differences were statistically significant (F=127.43 and 21.10,both P＜0.05).The positive percentage of CCR6 protein expression in HCC tissues (54.17％,26/48) was significantly higher than that in para carcinoma tissues (16.67％,8/48) and normal liver tissues (0/8),and the difference was statistically significant (x2 =19.55,P＜0.05).There was no statistically significant difference in the positive percentage of CCL20 protein expression among HCC tissues (50.00％,24/48),paracarcinoma tissues (33.33％,16/48),normal liver tissues (2/8) (all P＜0.05).There was correlation between the positive percentage of CCR6 protein expression and that of CCL20 protein expression in HCC tissues (r=0.42,P＜0.05).The positive percentage of CCR6 protein expression was correlated with the pTNM stage of HCC,vascular tumor thrombosis,intrahepatic metastasis and lung metastasis (x2 =5.48,4.02,5.07 and 5.19,all P＜0.05).The positive percentage of CCL20 expression was significantly correlated to tumor maximum diameter and pTNM stage (x2 =4.09 and 4.00,both P＜0.05).Both the disease-free survival (DFS) rate and overall survival (OS) rate of CCR6-positive group were significantly lower than those of negative group (x2 =4.57 and 6.57,both P＜ 0.05).There were no significant differences in DFS rate and OS rate between CCL20-positive group and negative group (both P＞0.05).Conclusion CCR6/CCL20 axis may be related with the malignant behavior and the prognosis of HCC.
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Chemokine is a kind of cytokine that induces cell transport to inflammation sites. It plays an important role in controlling the differentiation, development and directed migration of immunocytes. CCL20 is one of the important members of chemokine family, and it belongs to CC subfamily. Its receptor is CCR6. CCL20 can be expressed in activated monocytes, T lymphocytes, dendritic cells and endothelial cells, and CCR6 can be expressed in liver, lung, and lymphoid tissues. The expression of CCL20 can be induced by tumor necrosis factor α (TNFα), interleukin 1β (IL-1β), IL-17, CD40 ligand and interferon γ (IFN-gamma;). In summary, CCL20 plays an important role in tumor growth, invasion and metastasis (mainly refers to intrahepatic metastasis). Copyright© 2011 by TUMOR.
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A doença enxerto contra hospedeiro (GVHD) é uma complicação comum nos pacientes submetidos ao transplante de células-tronco hematopoiéticas (TCTH), sendo considerada a maior causa de morbidade e mortalidade nesses pacientes. O principal objetivo do presente estudo foi relacionar a concentração de células de Langerhans em mucosa bucal de pacientes com GVHDc bucal com a expressão da quimiocina CCL20 e de seu receptor CCR6 no epitélio bucal, a fim de elucidar os mecanismos biológicos envolvidos no recrutamento das células de Langerhans na GVHDc. Foram selecionados fragmentos obtidos por biópsia de mucosa bucal de 60 pacientes onco-hematológicos e hematológicos submetidos previamente ao transplante de células tronco hematopoiéticas no Hospital Amaral Carvalho, Jaú SP, onde 30 pacientes desenvolveram GVHDc em mucosa bucal (Grupo 1) e 30 não desenvolveram GVHDc (Grupo 2). Amostras obtidas a partir de 30 biópsias de lesões não inflamatórias em mucosa bucal constituíram o Grupo Controle (Grupo 3). Cortes microscópicos foram avaliados em coloração de rotina Hematoxilina e Eosina, e submetidos à técnica imuno-histoquímica, utilizando-se anticorpos monoclonais anti-CD1a e anti-CCR6, e anticorpos policlonais anti-CCL20. As células de Langerhans CD1a+ foram quantificadas no epitélio da mucosa bucal, e os resultados demonstraram um maior número destas células nos pacientes com GVHDc quando comparados àqueles sem GVHDc e ao Grupo Controle (p<0,001). A análise da imunomarcação das moléculas CCR6 e CCL20 foi subjetiva com aplicação de escores. Quanto à molécula CCR6, houve maior expressão no Grupo 1 (p<0,001) em comparação aos outros Grupos; porém, quanto à expressão de CCL20, não houve diferença estatística entre os três Grupos (p=0,108). Estes resultados sugerem que o aumento das células de Langerhans, na doença enxerto contra hospedeiro crônica, em mucosa bucal, pode estar associado a maior expressão do receptor CCR6. Possivelmente, o maior recrutamento de células de...
The graft versus host disease (GVHD) is a common complication in patients undergoing hematopoietic stem cell transplantation (HSCT), and considered a major cause of morbidity and mortality in these patients. The main objective of this study was to compare the concentration of Langerhans cells in oral mucosa of patients with oral chronic GVHD (GVHDc) with the expression of the chemokine CCL20 and its receptor CCR6 in oral epithelium, in order to clarify the biological mechanisms involved in the recruitment of Langerhans cells in GVHDc. We selected 60 biopsies of oral mucosa from onco-hematological and hematological patients submitted to prior hematopoietic stem cell transplantation at Hospital Amaral Carvalho, Jaú - SP from which 30 patients developed GVHDc in the oral mucosa (Group 1) and 30 did not develop GVHDc (Group 2). The Control Group (Group 3) was obtained from 30 biopsies of non-inflammatory lesions of oral mucosa. Microscopic sections were evaluated in routine Hematoxylin and Eosin staining, and submitted to immunohistochemistry using anti-CD1a and anti-CCR6 monoclonal antibodies, and anti-CCL20 polyclonal antibody. The Langerhans cells (CD1a+) were quantified in the epithelium of the oral mucosa, and the results showed a greater number of these cells in patients with GVHDc compared to those without GVHDc and the Control Group (p<0.001). Analysis of immunostaining of molecules CCL20 and CCR6 were subjective with application of scores. The expression of CCR6 molecule was more significant in Group 1 (p<0.001) compared to other groups, but in relation to CCL20 expression, there was no statistical difference between the three groups (p=0.108). These results suggest that the increase of Langerhans cells in GVHDc affecting oral mucosa may be associated with increased expression of the receptor CCR6. We suggest that the increased recruitment of Langerhans cells to the oral mucosa in patients with transplanted bone marrow contributes...
Subject(s)
Humans , Male , Female , Adult , Langerhans Cells/pathology , Graft vs Host Disease/pathology , Mouth Mucosa/pathology , Chemokines/biosynthesis , /biosynthesis , Biopsy , Graft vs Host Disease/metabolism , Biomarkers/metabolism , Mouth Mucosa/metabolism , Sex Distribution , Statistics, Nonparametric , Hematopoietic Stem Cell Transplantation/adverse effectsABSTRACT
Objective To investigate the expression of CCL20 and CCR6 in the patients with gastric cancer and To examine the relationship between chemokine expression and the occurrence and development of Gastric Cancer. Methods Real-time PCR , flow cytometry and ELISA are used to measure the gene transcription and protein expression levels of chemokine CCL20 and CCR6 in the serum of 50 patients with Gastric Cancer and 30 normal controls. Results The gene expression levels CCL20 and CCR6 in Gastric Cancer group are significantly higher than that in healthy controls. The level protein of CCL20 and CCR6 in peripheral blood of patients with gastric cancer are significantly higher than that in healthy peep le[ (45.4 ±10.9) pg/mL vs (18.6±4.7) pg/mL; (7.11 ±1.03%) vs (1.83±0.43%), P＜0.01. respectively],and the increase significantlyassociated with the clinical stage of Gastric Cancer. Conclusions The method for detecting the expression of CCL20 and CCR6 in patient with Gastric Cancer has been successfully established, and their expression levels were found to be correlated with the occurrence and development of Gastric Cancer. Thus, CCL20 and CCR6 may be involved in the regulatory mechanisms associated with the development of Gastric Cancer, and may be valuable in its diagnosis and prevention.
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Objective To observe the correlation of histologicalactivity(HAI) of chronic hepatitis B (CHB) with CCL20 expression, and to investigate the impact of CCL20 expression in CHB infection. Methods On the basis of established competitive quantitative RT-PCR with an internal standard, the expression of the CCL20 in the hepatocytes in different infected patterns of HBV infected cells and liver biopsies were quantified and at the same time its correlation to HAI were explored. Results In the cell levels, the expression quantity of CCL20 in control cells (HepG2), persistent HBV infected hepatocytes( HepG2. 2. 15) are (2. 65 ± 0. 02) pg/106 cells, ( 1.22± 0. 04) pg/106 cells, respectively. There were significantly differences between them ( t = 39. 66, P < 0. 01 ). The expression of CCL20 was enhanced in hepatocytes stimulated by PMA but their expression pauern was not changed. Moreover, CCL20 expression in liver biopsies with CHB was (3.54 ± 0. 65 ) pg/20 mg and CCL20 expression in control groups was ( 8. 74±0. 56) pg/20 mg. The expression of CCL20 between two groups was different (t =30. 09,P <0. 01 ) and correlation lied in between HAI and CCL20 expression in liver biopsies of CHB patients ( r = 0. 675, P =0. 023 ). Conclusion CCL20 expression was down-regulated and it was correlated to HAI of liver biopsies in CHB patients.