Research progress of CCL20-CCR6 axis and epithelial-mesenchymal transition in breast cancer / 国际外科学杂志

CCL20 and CCR6 are chemokines produced by a variety of cells. CCL20 and CCR6 combine to stimulate a series of downstream pathways, participate in the occurrence and development of various malignant tumors, and also play an important role in the invasion and metastasis of breast cancer and the process of chemotherapy resistance. Epithelial-mesenchymal transformation (EMT) is a key step in the process of tumor cell metastasis, which is characterized by loss of cell adhesion, down-regulation of E-cherherin expression, up-regulation of mesenchymal markers and fibrinectin expression, and enhancement of cell motor ability and invasion ability. This article reviews the research of CCL20-CCR6 biological axis and EMT on invasion and metastasis of breast cancer cells.

Lymphocyte B and Th17 chemotactic cytokine levels in peri-implant crevicular fluid of patients with healthy, peri-mucositis, and peri-implantitis implants / Niveles de citoquinas quimiotácticas de linfocitos B y Th17 en el fluido crevicular periimplantario de pacientes con implantes sanos, perimucositis y periimplantitis

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.

AKT/MMP3 activation, not EMT pathway, participates in CCL20-induced invasion and migration of colon cancer SW480 cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the molecular mechanism of chemokine CCL20/CCR6 in promoting invasion and migration of colon cancer SW480 cells. Methods: Colorectal cancer SW480 cells with high expression of CCR6 receptor were screened by immunochemistry (IHC). After co-culture with recombinant human CCL20, the invasion and migration of SW480 cells were detected by Transwell assay and Wound-Healing assay, respectively. Expressions of EMT markers, AKT signal protein and target protein MMP3 were detected by immunofluorescence (IF) and WB. AKT signaling pathway as the key mechanism was confirmed by MK2206 blocking assay. The expressions of CCL20 and MMP3 in colorectal cancer tissues as well as their correlation were analyzed by TCGAdatabase resources (https://portal.gdc.cancer.gov/). Results: CCL20 promoted the invasion and migration ability of SW480 cells significantly (all P <0.01), and this was induced by activation of AKT signaling and up-regulation of downstream target protein MMP3, instead of EMT. Blocking AKT signaling could significantly inhibit the invasion and migration of SW480 cells, and down-regulate MMP3 expression (P<0.05). TCGA platform data showed that the expressions of CCL20 and MMP3 in colorectal cancer tissues were significantly higher than those in normal mucosa tissues (P<0.05 or P<0.01), and an evidently positive correlation was found between CCL20 and MMP3 (r =0.051, P<0.01). Conclusion: The chemokine CCL20 promotes the invasion and migration of SW480 cells throughAKT/MMP3 signal axis, but not the EMT.

In vitro chemokine (C-C motif) receptor 6-dependent non-inflammatory chemotaxis during spermatogenesis

BACKGROUND: Chemokine (C-C motif) receptor 6 (CCR6) is present in sperm and plays a significant role in sperm motility and chemotaxis acting in the reproductive tracts. However, the expression and functional significance of CCR6 in testis are still poorly understood, especially in the process of spermatogenesis. METHODS AND RESULTS: CCR6 was expressed in spermatogenic cell lines and its expression was shown in an age-dependent upregulation manner from puberty to adulthood in mouse testis. Immunostaining results confirmed the localization of CCR 6 in testis. Further chemotaxis assays demonstrated that spermatogenic cells GC-1 and -2 exhibited a directional movement toward CCR6-specific ligand such as CCL20 or Sertoli cells in vitro. CONCLUSIONS: The present findings indicate that CCR6 is involved in the chemotaxis of spermatogenic cells in vitro and promotes chemotaxis under non-inflammatory conditions during normal spermatogenesis.

Research advances in the role of CCL20/CCR6 in liver cancer and other tumors / 临床肝胆病杂志

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.

Effects of interleukin-36ot on psoriasiform skin lesions and C-C motif chemokine ligand 20 expression in mice / 中华皮肤科杂志

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.

Effects of narrow-band ultraviolet B on levels of plasmin and CC chemokine ligand 20 in peripheral blood of patients with psoriasis vulgaris / 中华皮肤科杂志

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.

Construction and expression of recombinant adenovirus vector Ad5-CCL20 / 中国现代普通外科进展

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.

Effect of chemokine CCL20 and CCL22 combined with skin antigen-induced Treg on survival time of grafted skin / 中国免疫学杂志

Objective:To study the effect of chemokines CCL20 and CCL22 combined with skin-induced Treg on survival time of grafted skin.Methods: Skin grafting mice were divided into four groups, three mice per group, namely Treg group, Treg+CCL20 group,Treg+CCL22 group and control group.C57BL/6 mice were used as donor and BALB/c as acceptor, and the Treg cells were isolated from the mice induced by skin allograft.After skin grafted,CCL20 and CCL22 were subcutaneous injection every day,which lasted for 10 day.Survival time of skin in each group were observed and recorded.The Treg colonzation experiments were performed as follows.We firstly isolated Treg with Magnetic cell sorting system( MACS) and then labled them with 99 Tcm.After that we intravenously injected them into the mice.3 hours later,the mice were sacrifced and the radioactivity of organs were detected by GC-2016γradioim-munoassay counter.Results:①After Treg treated the survival time of skin grafted in antigen-induced Treg group was signifiantly longer than control group,when treg were cooperated with CCL20 and CCL22,the skin grafted showed more longer survival time than Treg and control groups( P<0.001 ).②After injection of induced Treg, Treg in autologous and allogeneic skin grafts goups were mainly distributed in autologous and allogeneic skin,accounting for 60% and 98% respectively.When cooperated with CCL20 or CCL22,the Treg were mainly distributed in liver.Conclusion:Chemokines CCL20 and CCL22 synergistically improved the effects of skin antigen induced Treg on survival time of skin graft,which probably related with the Treg colonization into the liver.

Role and recruitment of Th9 cells in liver cirrhosis patients

Objective: To investigate the role of T helper 9 (Th9) cells in liver cirrhosis (LC) patients and whether chemokine receptor type 6 (CCR6)/chemokine ligand 20 (CCL20) axis is involving in the recruitment of Th9 cells into liver. Methods: Peripheral blood and liver tissue from 30 LC patients and 18 normal controls were recruited. The frequency of Th9 cells and CCR4, CCR6 in the peripheral blood was tested by flow cytometry. Serum interleukin (IL)-9 and CCL20 levels were tested by enzyme-linked immunosorbent assay. Immunohistochemical staining was used to detect α-smooth muscle actin, CCR6 and CCL20 expression in liver tissue. Results: The frequency of Th9 cells in LC patients was significantly increased compared with controls (P < 0.05). The serum IL-9 level and CCL20 level increased markedly in LC patients compared with controls (P < 0.05), and IL-9 was positively correlated to Th9 cells and CCL20. Furthermore, the frequency of Th9 cells was correlated to prothrombin time, total bilirubin level, hyaluronic acid and type IV collagen in LC patients. We also found that Th9 cells in LC patients expressed higher frequency of CCR4

Effects of rottlerin on in vitro proliferation of and expressions of interleukin-17C, CCL20 and nuclear factor-κB in a human keratinocyte cell line HaCaT / 中华皮肤科杂志

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.

IL-17 expression in lesions of oral lichen planus and its auxo-action for chemokine CCL20 expression / 实用医学杂志

Objective To investigate the mechanism of IL-17, the signature cytokine produced by Th17 cells, in OLP lesion. Methods 24 patients with reticular OLP, 19 patients with atrophic-erosive OLP and 13 healthy volunteers were enrolled in this study . Real-time quantitative PCR ( real-time qPCR ) was performed to analyze the expressions of the production of IL-17 and CCL20 mRNA. Results The expressions of IL-17 mRNA in reticular OLP and atrophic-erosive OLP were significant higher than that in healthy oral mucosa (P = 0.0095, P <0.0001, respectively), meanwhile, remarkable increased IL-17 expression in atrophic-erosive OLP group was found compared with reticular OLP group (P = 0.0012). Additionally, the expressions of CCL20 mRNA in reticular OLP and atrophic-erosive OLP were significant higher than that in control group (P=0.0357, P<0.0001, respectively), meanwhile, CCL20 expression in atrophic-erosive OLP was higher than that in reticular OLP. The expressions of CCL20 mRNA rises with the increased expression of IL-17, and were positive correlated with IL-17 expressions in OLP lesions (P=0.003). Conclusions IL-17 production can induce chemokine CCL20 expression in OLP lesion. The signal pathway may promote the migration and infiltration of inflammatory cells in OLP lesions.

Expression and its clinical significance of chemokine receptor 6/chemokine ligand 20 axis in hepatocellular carcinoma / 中华消化杂志

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.

Doença enxerto contra hospedeiro crônica em mucosa bucal: relação da concentração de células de Langerhans com a expressão da quimiocina CCL20 e de seu receptor CCR6 / Chronic graft versus host disease in the oral mucosa: concentration of Langerhans cells and its relationship with the chemokine CCL20 and its receptor CCR6

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...

Chemokine CCL20 and malignant tumors / 肿瘤

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.

Clinical significance of chemokine CCL20 and its receptor CCR6 expression in patients with gastric cancer / 国际外科学杂志

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.

Significance of CCL20 expression in liver biopsies of chronic hepatitis B patients / 中华检验医学杂志

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.

Identification and Functional analysis of Gene Expression in Mycobacterium tuberculosis-infected Human Monocytic Cells Under Hypoxic Conditions

Mycobacterium tuberculosis-induced granulomatous lesions, particularly those undergoing central caseation, are known as hypoxic. To analyze the host genes associated with hypoxic conditions from cells infected with M. tuberculosis, we performed GeneChip analyses on mRNA from M. tuberculosis H37Rv-treated human monocytic THP-1 cells cultured in oxygen-depleted status for 18 h. The expression of 99 genes was altered, including those involved in intracellular signaling, energy production, and protein metabolism, as revealed by stringent microarray data analysis. Most notably, mRNA expression of chemokine macrophage inflammatory protein 3alpha/CC chemokine ligand 20 (CCL20) was significantly up-regulated in M. tuberculosis-infected cells under hypoxic conditions. We further analyzed the CCL20 expression in peripheral blood mononuclear cells (PBMCs) and monocyte derived macrophages (MDMs) from healthy controls and TB patients. A comparative analysis has revealed that the mRNA and protein expression of CCL20 were prominently up-regulated in PBMCs, and MDMs from TB patients, compared with healthy controls. Collectively, these data show that the gene expression of CCL20 was up-regulated in M. tuberculosis H37Rv-infected human monocytic THP-1 cells cultured in hypoxic conditions. In addition, the production of CCL20 is substantially increased in cells from TB patients than in healthy controls, suggesting an important role of CCL20 in the immunopathogenesis during TB infection.

Iron chelator inducesMIP-3alpha/CCL20 in human intestinal epithelial cells: implication for triggeringmucosal adaptive immunity

A previous report by this laboratory demonstrated that bacterial iron chelator (siderophore) triggers inflammatory signals, including the production of CXC chemokine IL-8, in human intestinal epithelial cells (IECs). Microarray-based gene expression profiling revealed that iron chelator also induces macrophage inflammatory protein 3 alpha (MIP-3alpha)/ CC chemokine-ligand 20 (CCL20). As CCL20 is chemotactic for the cells involved in host adaptive immunity, this suggests that iron chelator may stimulate IECs to have the capacity to link mucosal innate and adaptive immunity. The basal medium from iron chelator deferoxamine (DFO)-treated HT-29 monolayers was as chemotactic as recombinant human CCL20 at equivalent concentrations to attract CCR6+ cells. The increase of CCL20 protein secretion appeared to correspond to that of CCL20 mRNA levels, as determined by real-time quantitative RT-PCR. The efficacy of DFO at inducing CCL20 mRNA was also observed in human PBMCs and in THP-1 cells, but not in human umbilical vein endothelial cells. Interestingly, unlike other proinflammatory cytokines, such as TNF-alpha and IL-1beta, a time-dependent experiment revealed that DFO slowly induces CCL20, suggesting a novel mechanism of action. A pharmacologic study also revealed that multiple signaling pathways are differentially involved in CCL20 production by DFO, while some of those pathways are not involved in TNF-alpha-induced CCL20 production. Collectively, these results demonstrate that, in addition to some bacterial products known to induce host adaptive immune responses, direct chelation of host iron by infected bacteria may also contribute to the initiation of host adaptive immunity in the intestinal mucosa.

Effects of Sulphasalazine and Glucocorticoid on the Regulation of CCL20 Gene Expression in the Peripheral Blood Cells of Korean Patients with Ulcerative Colitis / 대한해부학회지

Discovery of Nod2 has brought to light the significance of mononuclear cells as well as epithelial cells in inflammatory bowel disease (IBD) pathogenesis. Similarly, CCL20 is expressed in both mononuclear cells and epithelial cells and is likely to link innate and acquired immunity. We therefore asked whether CCL20 expression is altered in the peripheral blood mononuclear cells (PBMCs) from patients with ulcerative colitis (UC), a major type of IBD in Korea, and is correlated with the disease activity. The expression levels of CCL20 mRNA were significantly high in the PBMCs from the patients with UC. CCL20 protein expression was also up-regulated in the mucosal epithelium in UC but not in normal controls. Interestingly, however, disease activity index (DAI) revealed that untreated UC groups express higher expression levels of CCL20 mRNA than treated UC groups, implying that CCL20 may be a potential target for the anti-inflammatory treatments. In an agreement with this, three months follow up study revealed that the UC patients who were treated with 5-amino salicylic acid (5-ASA) and glucocorticoid showed dramatic decrease in their CCL20 mRNA levels as compared to untreated ones. Moreover, TNF-alpha-or IL-1beta-induced CCL20 secretion in human epithelial HT-29 cells was significantly diminished by the treatment with 5-ASA and/or dexamethasone, suggesting that CCL20 may be one of the central targets of the anti-inflammatory drugs. Collectively, these results suggest that CCL20 expression in UC may be associated with altered immune and inflammatory responses in the blood as well as the intestinal mucosa and further implied a potential for CCL20 as an important diagnostic marker for UC.