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.

Expression of CCR7 and Tim-3 in Childhood Patients with Acute Lymphoblastic Leukemia and Their Predictive Value for Prognosis / 中国实验血液学杂志

OBJECTIVE@#To analyze the expression of CCR7 and Tim-3 in childhood patients with acute lymphoblastic leukemia (ALL) and their predictive value for prognosis.@*METHODS@#Eighty-six newly diagnosed ALL childhood patients from January 2007 to January 2017 treated in our hospital were selected. The expression level of CCR7 and Tim-3 in bone marrow isolated cells of ALL patients were detected by flow cytometry, all the patients were divided into the recurrence group and non-recurrence group according to the follow-up results, the differences in the expressions of CCR7, Tim-3 between the two groups were compared. The correlation between the expression of CCR7 , Tim-3 and the clinicopathologic features of ALL patients were analyzed, the predictive value of CCR7 and Tim-3 for the prognosis of newly ALL patients were evaluated by ROC curve, and the relationship between serum CCR7, Tim-3 and prognosis were analyzed.@*RESULTS@#The expression levels of CCR7 and Tim-3 in recurrence group were significantly higher than those in non-recurrence group(P0.05). The exogenous infiltration rate of patients with high expression of CCR7 and Tim-3 was significantly higher than those in low expression group (P<005). The high expression rate 76.9% of Tim-3 in patients with T-ALL was significantly higher than that of B-ALL patients with Tim-3 high expression rate 45.2% (P<0.05). The median OS of patients with CCR7 level ≥45.97% and <45.97% were 9.3 months and 13.6 months respectively(P=0.004), and the Tim-3≥53.54% and Tim-3<53.54% were 9.1 months and 13.6 months respectively(P=0.001). The results of Cox's multi-factor regression analysis showed that CCR7 level(HR=1.024, 95 CI 1.001-1.049) and Tim-3 level (HR=1.879, 95 CI 1.183- 2.985) were the independent risk factors that affect the OS in ALL patients(P<0.05).@*CONCLUSION@#The expression of CCR7 and Tim-3 in bone marrow isolated cells of ALL patients shows good predictive value for prognosis, and the combination of CCR7 and Tim-3 can improve the sensitivity of the detection, the higher expression of CCR7 and Tim-3 can be used as potential indexes in prognosis evaluate.

Expression and Clinical Significance of CC-chemokine Receptor 7 in Adult Acute Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the expression of CC-chemokine Receptor 7 (CCR7) in adult acute leukemia patients, and to analyze the relationship of CCR7 expression with the clinical characteristics of patients.</p><p><b>METHODS</b>The expression of CCR7 in bone marrow samples from adult acute leukemia patients were detected by flow cytometry (FCM), the relationship of CCR7 expression with the clinical characteristics of patients such as sex, age, WBC count, blast cell ratio, CD56 expression, molecular biology, cell genetics, risk stratification, extramedullary infiltration was analyzed.</p><p><b>RESULTS</b>The expression rate of CCR7 in adult ALL and AML patients was 36.8% and 9.6%, respectively, and the expression level of CCR7 in ALL patients was higher than that in AML patients (P < 0.05). The extramedullary infiltration rate was 100% and 41.7 % for CCR7 positive and negative groups of ALL, respectively (P < 0.05). While the mean fluorescence intensity (MFI) in extramedullary infiltration group of ALL was higher than that in none-extramedullary infiltration group of ALL (50.00 ± 10.42 vs 18.14 ± 1.39), respectively (P < 0.05).</p><p><b>CONCLUSION</b>CCR7 is higher expressed in adult acute leukemia cells, moreover its expression rate in ALL is higher than that in AML, and the expression of CCR7 is related with extramedullary infiltration in ALL.</p>

Effect of low-selenium diet on expressions of CCR7, CD206 and CD163 in the liver and kidney of rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of low-selenium diet on the liver and kidneys of rats and explore the role of macrophage polarization into M1 and M2 phenotypes in liver and kidney injuries.</p><p><b>METHODS</b>Twenty-four rats (12 female and 12 male) were randomly divided into control group and low-selenium group and fed with normal chow (dietary selenium of 0.18 mg/kg) and low-selenium diet (dietary selenium of 0.02 mg/kg) for 109 days. After the feeding, the rats were sacrificed for HE staining to observe liver and kidney pathologies, and immunohistochemistry was performed for analyzing CCR7, CD206, CD163-positive cell numbers in the liver and kidneys.</p><p><b>RESULTS</b>The rats in low-selenium group showed severer fibrosis in the liver and kidney than the control group. In either male or female rats in low-selenium group, CCR7 and CD206 expressions in the liver were comparable with those in control group, but CD163 expression was lower than that in the control group (P<0.05 for both female and male rats). In the kidney, the proximal tubule showed a slightly higher while the distal tubule showed a slightly lower CCR7 expression in low selenium group than in the control group (P>0.05). In low-selenium group, a significantly lower CD163 expression in the distal tubule and a significantly higher CD206 expression in the proximal tubule were noted as compared with the control group (P<0.05 in both female and male rats). Compared with the control rats, the male rats in low-selenium group, but not the female rats, showed a significantly lower CD163 expression in the proximal tubule of the kidney (P<0.05); the female but not the male rats in low-selenium group show a higher CD206 expression in the distal tubule (P<0.05).</p><p><b>CONCLUSION</b>Low-selenium diet can cause liver and kidney fibrosis in rats and may inhibit macrophage activation into the M2 phenotype.</p>

Effect of CO₂ pneumoperitoneum on the expression of the chemokine receptors CXCR4 and CCR7 in colorectal carcinoma cells in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The ability of pneumoperitoneum in laparoscopic surgery to promote proliferation and metastasis of colorectal cancer has become a focus of research in the field of minimally invasive surgery. The aim of this research was to investigate the effect of CO2 pneumoperitoneum under different pressures and exposed times on the expression of chemokine receptors in colorectal carcinoma cells.</p><p><b>METHODS</b>We constructed an in vitro pneumoperitoneum model. SW480 colon carcinoma cells were exposed to CO2 pneumoperitoneum under different pressures (6, 9, 12, and 15 mmHg) for 1, 2, and 4 hours. These cells were then cultivated under the same conditions as normal SW480 colon carcinoma cells without CO2 pneumoperitoneum (control group), treated at 37°C, and 5% CO2. The expression of the chemokine receptors CXC receptor 4 (CXCR4) and chemokine C receptor 7 (CCR7) was detected by immunocytochemistry and reverse transcriptase polymerase chain reaction after being cultivated for 0, 24, 48, and 72 hours.</p><p><b>RESULTS</b>Immunocytochemistry showed that CXCR4 expression in SW480 cells was significantly decreased in the 6, 9, 12, and 15 mmHg CO2 pneumoperitoneum-treated groups for the same exposure times compared with controls (P < 0.05). CCR7 expression in SW480 cells was significantly decreased in the 12 and 15 mmHg CO2 pneumoperitoneum-treated groups compared with controls (P < 0.05). CXCR4 and CCR7 expression increased up to the level of the control group after 24 and 48 hours (P > 0.05). If the CO2 pneumoperitoneum pressure increased, CXCR4 and CCR7 expression decreased at all exposure times. If the CO2 pneumoperitoneum exposure time prolonged, there were no significant differences in CXCR4 and CCR7 expression under the same pressure. Under all exposure times, CXCR4 and CCR7 mRNA expression was significantly decreased in the 6, 9, 12, and 15 mmHg CO2 pneumoperitoneum-treated groups (P < 0.05) compared with controls, and it increased up to the level of controls after being cultivated for 48 hours (P > 0.05). If the CO2 pneumoperitoneum pressure increased (with all exposure times) and exposure time prolonged (under the same pressure), there were no significant differences in CXCR4 and CCR7 expression.</p><p><b>CONCLUSIONS</b>CXCR4 and CCR7 expression is temporarily affected after continuous CO2 pneumoperitoneum treatment. The high pressure of CO2 pneumoperitoneum plays an important role in suppressing the expression of these chemokine receptors. Different lengths of time of exposure to a CO2 pneumoperitoneum-like environment do not change CXCR4 and CCR7 expression.</p>

Effects of immature dendritic cells to express CCR7 on graft-versus-host disease in allogeneic bone marrow transplant mouse model / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of immature dendritic cells (imDC) expressing chemokine receptor-7 (CCR7) on acute graft-versus-host disease (aGVHD) in allogeneic bone marrow transposed (allo-BMT) mouse model.</p><p><b>METHODS</b>We constructed the lentiviral vectors carrying mouse CCR7 gene and infect imDC effectively in vitro. GVHD model was established with C57BL/6(H-2b) donor mice and BALB/c (H-2d) recipient mice. After irradiation, recipients were injected with donor bone marrow and spleen cells along with CCR7-modified dendritic cells. Mice were randomized into irradiation, transplant control, pXZ9-imDC (empty vector control) and CCR7-imDC groups. Survival, GVHD score, histopathological analysis and plasma levels of inflammatory cytokines were observed.</p><p><b>RESULTS</b>The mean survival in irradiation, transplantation, pXZ9-imDC and CCR7-imDC groups were (8.20±1.48)d, (12.20±2.78)d, (20.70±6.01)d and (27.5±7.55)d respectively. The survival in CCR7- imDC group was significantly improved compared with other groups (P<0.05). GVHD scores in transplantation, pXZ9-imDC and CCR7-imDC groups were (6.90±1.66), (5.60±0.97) and (4.10±1.79) respectively. CCR7-imDC group had significantly lower GVHD score and minor tissue damages shown by histopathological analysis than the other groups. Plasma IFN-γ level increased and reached the peak at +10 day in transplant group, while it gradually decreased in pXZ9-imDC and CCR7-imDC groups, and then reached the nadir at +20 day post-allo-BMT, with the lowest level in CCR7-imDC group (P<0.01). Plasma IL-4 decreased in transplant group, while it gradually increased in pXZ9-imDC and CCR7-imDC groups and reached the highest level at + 10 day in CCR7- imDC group (P<0.01). The 95%-100% of H-2b positive cells in recipient mice on + 30 day post-allo-BMT demonstrated the complete donor- type implantation.</p><p><b>CONCLUSION</b>Genetically modified immature DC by CCR7 gene could alleviate damages by GVHD and prolong survival of recipient mice after allo-BMT.</p>

Influence of infection of murine chemokine receptor-7 recombinant lentivirus on the immunogenicity and migration of DC 2.4 cells / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the influence of infection of murine chemokine receptor-7 recombinant lentivirus on the immunogenicity and migration of dendritic cell strain DC 2.4 cells.</p><p><b>METHODS</b>DC 2.4 cells were routinely cultured. Lentiviruses carrying GFP and those with up-regulated CCR7 were constructed. DC 2.4 cells were divided into DC 2.4 group (without any treatment), GFP-DC 2.4 group (infected with GFP-carrying lentivirus), and CCR7-DC 2.4 group (infected with CCR7-carrying lentivirus labeled by GFP) according to the random number table. The expressions of surface molecules MHCII, CD80, CD86, and CCR7 were detected by flow cytometry, Western blotting, and confocal laser scanning microscope. The migration of cells was detected by chemotaxis assay in vitro. The immunogenicity of cells was detected with mixed lymphocyte reaction. LPS-DC 2.4 group was set up as positive control. Data were processed with one-way analysis of variance and t test.</p><p><b>RESULTS</b>Lentiviruses carrying stably-expressing CCR7 were constructed, and the transfection rate of which into DC 2.4 cells was 87.4%. There was no statistically significant difference among DC 2.4, GFP-DC 2.4, and CCR7-DC 2.4 groups in the expressions of MHC II, CD80, and CD86 as showed by flow cytometry (with F values from 0.17 to 1.19, P values all above 0.05). The protein expression of CCR7 of cells in CCR7-DC 2.4 group (45.1 ± 2.1) was obviously higher than that in DC 2.4 and GFP-DC 2.4 groups (25.3 ± 1.4, 28.6 ± 0.9, F = 162.90, P < 0.01), while the difference of which between DC 2.4 group and GFP-DC 2.4 group was not statistically significant (t = 2.20,P > 0.05). The fluorescence intensity of CCR7 in CCR7-DC 2.4 group was obviously increased compared with that of DC 2.4 group. The chemotaxis migration rate of cells in CCR7-DC 2.4 group with the influence of CCL19 was (41.0 ± 2.0)%, which was significantly higher than that of DC 2.4 and GFP-DC 2.4 groups [(6.0 ± 0.5)%, (6.8 ± 0.3)%, F = 84.21, P < 0.01]. There was no statistically significant difference between DC 2.4 group and GFP-DC 2.4 group in the migration rate (t = 0.45, P > 0.05). The absorbance values in DC 2.4, GFP-DC 2.4, CCR7-DC 2.4, and LPS-DC 2.4 groups were respectively 1.6 ± 0.4, 1.9 ± 0.4, 1.7 ± 0.4, 3.8 ± 0.4, and the differences among the former three groups were not obvious (F = 1.56, P > 0.05). The absorbance value in LPS-DC 2.4 group was obviously higher than that of the other three groups (with t values from 1.53 to 1.82, P values all below 0.01).</p><p><b>CONCLUSIONS</b>DC 2.4 cells infected with efficiently CCR7-expressing lentivirus showed high chemotaxis to CCL19, but without obvious change in immunogenicity.</p>

Expression of CCR6 and CCR7 in laryngeal squamous cell carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To evaluate the expressions of chemokine receptor 6 (CCR6), chemokine receptor 7 (CCR7) and their ligands (CCL20, CCL19/CCL21) in laryngeal squamous cell carcinoma (LSCC), and then explore their correlation with the clinicopathological features of LSCC.@*METHOD@#Blood samples, fresh specimens of LSCC and paired adjacent tissues were collected. The expressions of CCR6, CCR7 and their ligands CCL20, CCL19/ CCL21 mRNA as well as the protein CCR6, CCR7 were detected by real-time qRT-PCR and IHC respectively. Flow cytometry was also used to investigate CCR6, CCR7 expressed on PBMC.@*RESULT@#The relative expression levels of CCR6, CCR7, CCL19 and CCL21 mRNA in tumor tissue was significantly higher than that of adjacent tissues (P < 0.05), while the relative expression level of CCL20 mRNA in tumor tissue were significantly lower than that of adjacent tissues (P < 0.05). IHC confirmed the expression of protein CCR6 and CCR7 in both tumor tissue and metastatic ILN and the expression levels of protein CCR6, CCR7 were higher in the cases with lymphatic metastasis than that of those without lymphatic metastasis (P < 0.05). FCM showed the percentage of CD4+ CCR6+ T cells of LSCC was significantly higher than that of normal control (P < 0.05), while that of CD4+ CCR7+ T cells was significantly lower (P < 0.05).@*CONCLUSION@#CCR6 and CCR7 are expressed in tumor situ, metastatic LN and PBMC,and might exert a potential role in LSCC development.

Detection and its clinical value of CCR5 and CCR7 in dendritic cells from patients with active rheumatoid arthritis / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expressions of CCR5 and CCR7 on dendritic cells (DCs) in patients with rheumatoid arthritis (RA) in different phases of disease activity, and explore the relationship between the disease activity and the expression of chemokine receptors.</p><p><b>METHODS</b>Twenty-eight patients with low, moderate and high disease activity and 10 normal control subjects were enrolled in this study. Peripheral blood was obtained from the subjects and the DCs were isolated. The expression of CCR5 and CCR7 on DCs were detected by flow cytometry, and the serum levels of rheumatoid factor (RF), C-reactive protein (CRP) and anti-CCP antibody (ACPA) were assessed. The correlation of the expressions of CCR5 and CCR7 to serum RF, CRP, and ACPA levels of the RA patients were analyzed.</p><p><b>RESULTS</b>Compared to the normal control group, RA patients showed enhanced expressions of CCR5 and CCR7 on the DCs. A linear correlation was noted between CCR5 and CCR7 expressions on the DCs and the serum levels of RF and CRP, but not ACPA, in the RA patients.</p><p><b>CONCLUSION</b>The expressions of CCR5 and CCR7 on the DCs may correlate to the disease activity of RA, and may serve as valuable indices in monitoring the disease activity and the efficacy of the treatment.</p>

Expression and clinical significance of CCR6, CCR7 and CD4(+)CD25(+) Foxp3(+) regulatory T cells in laryngeal squamous cell carcinoma and neck lymphatic metastasis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the expressions and clinical significance of chemokine receptor 6 (CCR6), chemokine receptor 7(CCR7) and CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg) in laryngeal squamous cell carcinoma (LSCC) and metastatic lymph nodes.</p><p><b>METHODS</b>Blood samples and fresh specimens of LSCC were obtained from 50 LSCC patients treated. Blood samples from normal subjects were donated by 20 volunteers. The mRNA expressions of CCR6, CCR7 and their ligands CCL20, CCL19/CCL21 were detected by real-time quantitative RT-PCR. Expressions of CCR6 and CCR7 proteins were examined with immunohistochemistry. Flow cytometry was used to investigate Treg in peripheral blood mononuclear cells.</p><p><b>RESULTS</b>The relative expression levels of CCR6, CCR7 and CCL19 mRNA in tumor tissues with lymphatic metastases were significantly lower than those without lymph node metastases (P < 0.05), while the relative expression level of CCL20 mRNA in tumor tissues with lymphatic metastases was significantly higher than that of those without metastatic lymph nodes (t = 2.39, P < 0.05). Immunohistochemistry showed that the expression levels of CCR6 and CCR7 in tumor tissues in patients with lymph node metastases than those without lymph node metastases. CCR6 and CCR7 expressions were also detected in metastatic lymph nodes. Flow cytometry showed that the percentage of Treg in blood in LSCC patients was significantly higher than that in normal subjects (t = 2.19, P < 0.05), and those with lymph node metastasis had a much higher percentage of Treg (t = 2.14, P < 0.05).</p><p><b>CONCLUSIONS</b>CCR6, CCR7 and Treg could take part in the process of lymph node metastasis in LSCC patients.</p>

Effect of chemokine receptor 7 small interfering RNA on proliferation and invasion of squamous cell carcinoma of head and neck / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the effect of chemokine receptor 7 (CCR-7) small interfering RNA (siRNA) on proliferation and invasion of squamous cell carcinoma of head and neck (SCCHN).</p><p><b>METHODS</b>CCR-7 siRNA was co-transfected into SCCHN cell line PCI-4B using Lipofectamine 2000. CCR-7 protein level was detected by western blotting. SCCHN cell proliferation was detected by MTT, and the change of actin cytoskeleton observed by confocal laser scanning microscope. Transwell assays were used to determine chemotaxis and invasion of SCCHN cells. The activity and nuclear translocation of nuclear factor-kappa B (NF-kappa B) were detected by TransAM NF-kappa B p65 kit and fluorescence microscope respectively.</p><p><b>RESULTS</b>After CCR-7 siRNA transfection, the protein level of CCR-7 was significantly decreased. The changes induced by CCL-19, including increased proliferation rate, polarized actin polymerization, increased chemotaxis rate and invasion rate, were all abolished by CCR-7 siRNA transfection. CCR-7 siRNA also diminished CCL-19-induced NF-kappaB activation and nuclear translocation.</p><p><b>CONCLUSIONS</b>CCR-7 siRNA could inhibit expression of CCR-7 and diminish the increased proliferation and invasion of SCCHN induced by CCL-19 in vitro. CCR-7 siRNA may provide a potential treatment strategy for SCCHN.</p>

Influences of interferon-alpha on expression of Th cytokines and CCR7 in dendritic cells from patients with chronic myeloid leukemia in vitro / 中国实验血液学杂志

This study was aimed to investigate the influences of interferonalpha (IFN-alpha) on expressions of CCR7, interleukin10 (IL-10) and IL-12p70 in dendritic cells (DCs) from patients with chronic myeloid leukemia (CML). In addition to stem cell factor (SCF), granulocyte-macrophage colony stimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-alpha) and IL-4, IFN-alpha was added to the serum-free medium of DCs. After culture for 10-14 days, phenotypes and function of CML-DCs were evaluated respectively by flow cytometry and methyl thiazolyl tetrazolium (MTT) assay. Chromosome of DCs was analyzed by displaying G banding assay. The concentrations of IL-10 and IL-12P70 in supernatants were evaluated by enzyme-linked immunosorbent assay (ELISA). The results showed that the expressions of CD40, CD83, CD86 and CCR7 and the OD value in allogeneic mixed-lymphocyte reaction (MLR) in group with IFN-alpha (300 U/ml) were twice as high as those in group without IFN-alpha. The percentage of Ph1 positive cells and concentrations of IL-10 and IL-12 P70 were reduced in group with IFN-alpha. It is concluded that the defective phenotypes and functions of CML-DCs can be recruited partly by IFN-alpha. The mechanism may lie in the facts that expression of CCR7 and co-stimulatory molecules is promoted and the inhibitory effect of IL-10 on CML-DCs is relieved partly through the regulation of IFN-alpha.

Expression of chemokine receptor-7 in oral squamous cell carcinoma and its relation to cervical lymph node metastasis / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression of chemokine receptor CCR7 in oral squamous cell carcinoma (OSCC) and its possible role in directional neck lymph node metastasis.</p><p><b>METHODS</b>The expression of chemokine receptor CCR7 was examined in 85 cases of OSCC, normal oral mucosa, and Tca8113 and adenoid cystic carcinoma (ACC-M) cell lines using immunohistochemistry, RT-PCR and Western blotting. The relationship between the expression and clinicopathological factors of OSCC was analyzed. After treatment with or without CCR7 antibody, in vitro adhesion assay was performed to compare the adhesion of Tca8113 and ACC-M cells to lymph nodes.</p><p><b>RESULTS</b>CCR7 was expressed in 66% (56/85) of OSCC cases, and there was a significant difference in CCR7 expression between metastasis group (31/39, 79%) and non-metastasis group (25/46, 54%). RT-PCR and Western blotting also confirmed the presence of positive CCR7 expression in OSCC. Tca8113 cells expressing CCR7 showed stronger ability to adhere to lymph nodes as compared to CCR7-negative ACC-M cells and could be actively inhibited by CCR7 antibody.</p><p><b>CONCLUSIONS</b>CCR7 may play an important role in the development and lymph node metastasis of oral squamous cell carcinoma.</p>

Assignment of CCR7 gene to chicken chromosome 27 by radiation hybrid panel mapping / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The protein encoded by CC chemokine receptor 7 (CCR7) is a member of the G protein-coupled receptor family. This receptor was identified as a gene induced by the Epstein-Barr virus (EBV), and is thought to be a mediator of EBV effects on B lymphocytes. This receptor is expressed in various lymphoid tissues and activates B and T lymphocytes. It has been shown to control the migration of memory T cells to inflamed tissues, as well as stimulate dendritic cell maturation. To map the CCR7 gene in chicken chromosome, a 6 000 rads chicken-hamster radiation hybrid panel (ChickRH6) was used. PCR of samples from ChickRH6 revealed that the location of CCR7 gene is linked to the maker SEQ0347 (6 cR away) with LOD score of 16.6 and that the marker SEQ0347 is located on chromosome 27 at 27 cR of RH (radiation hydrid) map. We compared the corresponding human mRNA sequence with the predicted coding sequence of chicken CCR7 gene, and found that the assembled contig shared a high percentage of similarity with that of the human gene.

Implications of lung interstitial dendritic cells of mice in multiple organ dysfunction syndrome / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore the role of lung interstitial dendritic cells in immunodissonance and organ injury in multiple organ dysfunction syndrome (MODS).</p><p><b>METHODS</b>Animal model of MODS was established by injecting zymosan into the peritoneal cavity of C57BL/6 mice. The mice were randomly divided into groups of normal, 3 - 6 hours, 12 - 48 hours, 5 - 7 days, 10 - 12 days post injection. Pathological changes of lung and interstitial dendritic cells were studied by light and transmission electron microscope. Immunohistochemistry, RT-PCR and flow cytometry analyses were used to document status of biomarkers, including specific surface markers (CD205 and CD11c), costimulatory molecules (CD80 and CD86), SLC and its receptor CCR7 in lung, CD4+ and CD8+ T lymphocyte subtypes in peripheral blood.</p><p><b>RESULTS</b>At early stage of injury, interstitial dendritic cells showed an increase in proliferation with expression of low level of CD80 and CD86. In contrast, the expression of SLC and its receptor CCR7 in lung were increased. The ratio of CD4+/CD8+ declined in peripheral blood. At the stage of SIRS, interstitial dendritic cells continued to proliferate with high expressions of CD80 and CD86. SLC and CCR7 in lung also increased. The ratio of CD4+/CD8+ declined markedly in peripheral blood. At the MODS stage, interstitial dendritic cells further proliferated, but the expression of CD80 and CD86 declined to a very low level. Although the level of SLC increased consistently, the level of CCR7 continued to decrease, along with a markedly decreased CD4+/CD8+ ratio in peripheral blood.</p><p><b>CONCLUSIONS</b>Alterations of lung interstitial dendritic cells are likely to influence the course of immunological dysfunction of MODS. The level of CCR7 may serve as an indicator of the migration activity of interstitial dendritic cells and systemic immune response.</p>

Effects of mouse bone marrow mesenchymal stem cells on the expression of chemokine receptors of spleenocytes after phytohemagglutinin stimulation / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the effect of mouse bone marrow mesenchymal stem cells (MSCs) on the expression of chemokine receptors in T lymphocytes in vitro.</p><p><b>METHODS</b>Mouse bone marrow MSCs were separated with Percoll, cultured and expanded in low glucose DMEM. C57BL/6 mouse spleenocytes were cultured in the 24-hole flasks by the density of 1 x10(6)/hole. Phytohemagglutinin (PHA) was then added to the holes and cultured for 72 hrs. This study consisted of three groups. Groups A and B were co-cultured by adding MSCs as the ratio of 0.1 and 0.01 to spleenocytes respectively. The control group was cultured without MSCs. Three days later the suspended spleenocytes were harvested for detecting the expression of three chemokine receptors CXCR3, CCR5 and CCR7 in T lymphocytes by the flow cytometry.</p><p><b>RESULTS</b>The expression of CD3(+)CCR5(+) and CD3(+)CCR7(+) were statistically different among the three groups. Group A had the strongest expression, followed by group B and the control group. The expression of CD3(+)CXCR3(+) in group A was statistically higher than that in group B and the control group.</p><p><b>CONCLUSIONS</b>MSCs could up-regulate the expression of chemokine receptors CXCR3, CCR5 and CCR7 in T lymphocytes stimulated by PHA.</p>