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.
Cell Line, Tumor ; Chemokine CCL21 ; Endothelial Cells ; Humans ; Lymphatic Metastasis ; Neoplasms/therapy* ; Receptors, CCR7/genetics*

OBJECTIVETo 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.

METHODSDC 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.

RESULTSLentiviruses 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).

CONCLUSIONSDC 2.4 cells infected with efficiently CCR7-expressing lentivirus showed high chemotaxis to CCL19, but without obvious change in immunogenicity.

Animals ; Cell Line ; Cell Movement ; Dendritic Cells ; cytology ; immunology ; Lentivirus ; genetics ; Mice ; Receptors, CCR7 ; genetics ; metabolism ; Transfection

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.
Animals ; Base Sequence ; Chickens ; genetics ; Chromosomes ; genetics ; Humans ; Molecular Sequence Data ; Radiation Hybrid Mapping ; methods ; Receptors, CCR7 ; Receptors, Chemokine ; genetics ; Sequence Analysis, DNA ; methods ; Sequence Homology, Nucleic Acid

OBJECTIVETo 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.

METHODSThe 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.

RESULTSThe 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).

CONCLUSIONCCR7 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.

Adult ; Bone Marrow ; metabolism ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Leukocyte Count ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Receptors, CCR7 ; genetics ; metabolism

OBJECTIVETo 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).

METHODSCCR-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.

RESULTSAfter 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.

CONCLUSIONSCCR-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.

Carcinoma, Squamous Cell ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Head and Neck Neoplasms ; genetics ; pathology ; Humans ; Neoplasm Invasiveness ; RNA, Small Interfering ; Receptors, CCR7 ; genetics ; Transcription Factor RelA ; metabolism

Cadherins ; metabolism ; Female ; Gene Expression Profiling ; Humans ; Inflammatory Breast Neoplasms ; genetics ; metabolism ; pathology ; NF-kappa B ; metabolism ; Receptor, ErbB-2 ; metabolism ; Receptor, Notch3 ; Receptors, CCR7 ; metabolism ; Receptors, CXCR4 ; metabolism ; Receptors, Notch ; metabolism

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.
Cells, Cultured ; Dendritic Cells ; cytology ; Humans ; Interferon-alpha ; pharmacology ; Interleukin-10 ; genetics ; metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; immunology ; Philadelphia Chromosome ; drug effects ; Receptors, CCR7 ; genetics ; metabolism

OBJECTIVETo 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.

METHODSWe 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.

RESULTSThe 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.

CONCLUSIONGenetically modified immature DC by CCR7 gene could alleviate damages by GVHD and prolong survival of recipient mice after allo-BMT.

Animals ; Bone Marrow Transplantation ; adverse effects ; Dendritic Cells ; cytology ; Genetic Vectors ; Graft vs Host Disease ; prevention & control ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, CCR7 ; genetics ; Transplantation, Homologous