The study was aimed to explore the chemokine receptor CXCR4 expression on the B-lineage acute lymphocyte leukemia (B-ALL) cells of various differentiation stages and its relationship with myeloid antigen expression. Flow cytometry was used to detect the CXCR4 expression by means of double-fluorescence labeling with CD19/SCC gating. The results demonstrated that 92.9% B-ALL patients were positively expressed CXCR4. The CD10, CD34 antigens were differently expressed in differentiation stages of B-ALL. The immunotypes of (1) CD10(-)/CD34(+), (2) CD10(+)/CD34(+), (3) CD10(+)/CD34(-), (4) CD10(-)/CD34(-) presented at various differential stages from premature to mature. The positive rate of CXCR4 were (27.60 +/- 15.25)%, (30.95 +/- 15.50)%, (55.62 +/- 18.37)% and (77.25 +/- 10.86)% from (1) to (4) respectively. The median fluorescence intensity (MFI) of CXCR4 expression were 46.69 +/- 15.06, 47.43 +/- 12.39, 79.28 +/- 24.71 and 132.92 +/- 88.09. CXCR4 expressions were not significantly different between the premature stages of CD10(-)/CD34(+) and CD10(+)/CD34(+) subtypes, but both were lower than the CXCR4 expression in CD10(+)/CD34(-) and CD10(-)/CD34(-) subtypes. The highest incidence of CXCR4 expression was found in CD10(-)/CD34(-) B-ALL. The average level of CXCR4 expression on B-ALL cell with positive myeloid antigen CD13 or/and CD33 (my(+)B-ALL) was (12.56 +/- 3.88)% of positive rate and 39.82 +/- 11.58 of MFI, both of which were less than the positive rate (37.57 +/- 11.59)% and the MFI (50.72 +/- 13.34) on B-ALL cells with negative myeloid antigen expression (mye(-)B-ALL). In conclusion, the CXCR4 expression is associated with differentiation level of B-ALL cells and down-regulated when co-expressed with myeloid antigens.
Adolescent ; Adult ; Aged ; Antigens, CD34 ; analysis ; Burkitt Lymphoma ; metabolism ; pathology ; Cell Differentiation ; Child ; Female ; Humans ; Male ; Middle Aged ; Neprilysin ; analysis ; Receptors, CXCR4 ; analysis

OBJECTIVETo study the expression of stromal cell derived factor-1(SDF-1) and CXC chemokine receptor 4 (CXCR4) in the airway and the effect of budesonide on their expression in mice with asthma.

METHODSThirty BALB/c male mices were randomly divided into three groups: placebo control, untreated asthma, and budesonide-treated asthma. The asthma group were induced by intraperitoneal injection of 10% ovalbumin (OVA ) on days 1, 8 and 15, and then from days 22 to 34, challenged by inhalation of 2% OVA aerosol every other day. The budesonide-treated asthma group received an inhalation of budesonide (1 mg ) before OVA challenge. The pathological changes of the airway were assessed by hematoxylin and eosin staining. The immunohistochemistry was used to estimate the expression of SDF-1 in the lung. RT-PCR was used to evaluate the expression of CXCR4 in the lung.

RESULTSCompared with the control group, SDF-1 and CXCR4 expression in the lung in the untreated asthma group increased significantly (p<0.05). The budesonide-treated asthma group demonstrated significantly decreased SDF-1 (0.426+/-0.052 vs 0.361+/-0.065; p<0.05) and CXCR4 (0.829+/-0.027 vs 0.723+/-0.094; p<0.05) expression in the lung as compared with the untreated asthma group. Both SDF-1 (r=0.744, p<0.01) and CXCR4 (r=0.553, p<0.01)were positively correlated with the thickness of the airway wall.

CONCLUSIONSSDF-1 and CXCR4 may be associated with airway remodeling in mice with asthma. Budesonide can improve airway remodeling, possibly by decreasing the expression of SDF-1 and CXCR4.

Airway Remodeling ; drug effects ; Animals ; Asthma ; drug therapy ; metabolism ; pathology ; Budesonide ; pharmacology ; Chemokine CXCL12 ; analysis ; Male ; Mice ; Mice, Inbred BALB C ; Receptors, CXCR4 ; analysis ; genetics

BACKGROUNDIn a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explore the effect of CXCL12-CXCR4 axis on the metastasis of human ovarian cancer.

METHODSThe expressions of CXCR4 and CXCL12 mRNA and protein in human ovarian cancer cell line CAOV-3 was detected by RT-PCR and immunocytochemistry. Methythiazolyltetrazolium (MTT) was used to analyze the effect of different concentrations of CXCL12 on the proliferation of CAOV-3 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of CXCL12 and ascites on the migration and invasion of CAOV-3 cells. The expressions of integrin beta(1) and vascular endothelial growth factor-C (VEGF-C) mRNA were detected by RT-PCR. Data were analyzed using ANOVA by SAS 6.12.

RESULTSUnder serum-free suboptimal culture conditions, CXCL12 (100 ng/ml) significantly enhanced the proliferation of CAOV-3 cells compared with the control and 10 ng/ml CXCL12 groups (0.428 +/- 0.051 vs. 0.325 +/- 0.045 and 0.328 +/- 0.039, P < 0.05). This enhancing effect of CXCL12 was significantly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml CXCR4 antagonist AMD3100. However, 10 microg/ml neutralizing CXCR4 antibody could not inhibit cell proliferation without CXCL12. The levels of migration and invasion of the CAOV-3 cells treated with 100 ng/ml CXCL12 were significantly higher than those in the control (migration: 523.3 +/- 25.2 vs 108.0 +/- 7.2; invasion: 39.3 +/- 4.0 vs. 4.0 +/- 1.0). The enhancing effect of CXCL12 on cell migration and invasion increased with the concentration of CXCL12 (100 ng/ml vs10 ng/ml: migration, 523.3 +/- 25.2 vs 211.7 +/- 24.7; invasion, 39.3 +/- 4.0 vs 15.7 +/- 3.1, P < 0.05), and was strongly inhibited by 10 microg/ml neutralizing CXCR4 antibody or 1 microg/ml AMD3100. The number of migrated and invading cells in the CAOV-3 added with ascites was significantly higher than those in the 100 ng/ml CXCL12 group (migration: 706.6 +/- 30.6 vs 523.3 +/- 25.2, invasion: 61.7 +/- 7.6 vs 39.3 +/- 4.0, P < 0.05). The level of integrin beta(1) mRNA was greatly increased at 3 hours after being treated with CXCL12 (0.53 +/- 0.10 vs. 1.53 +/- 0.16, P < 0.05), and VEGF-C mRNA displayed significant augment at 24 hours after being treated with CXCL12 (0.52 +/- 0.09 vs 1.11 +/- 0.15, P < 0.05).

CONCLUSIONSCXCL12 and its receptor CXCR4 can promote the proliferation, migration, invasion of ovarian cancer cell line CAOV-3 and enhance its secretion of integrin beta(1) and VEGF-C. These effects can be inhibited by neutralizing CXCR4 antibody or AMD3100. CXCL12-CXCR4 axis plays an important role in ovarian cancer growth and metastasis.

Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Chemokine CXCL12 ; Chemokines, CXC ; analysis ; physiology ; Female ; Humans ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Ovarian Neoplasms ; pathology ; Receptors, CXCR4 ; analysis ; physiology

OBJECTIVETo explore the role and function of stromal cell-derived factor-1 (SDF-1) in stem cells migrating into injured brain area.

METHODSRat-derived nerve stem cells (NSCs) were isolated and cultured routinely. Transwell system was used to observe the migration ability of NSCs into injured nerve cells. Immunocytochemistry was used to explore the expression of chemotactic factor receptor-4 (CXCR-4) in NSCs. In vivo, we applied immunofluorescence technique to observe the migration of NSCs into injured brain area. Immunofluorescence technique and Western blotting were used to test expression level of SDF-1. After AMD3100 (a special chemical blocker) blocking CXCR-4, the migration ability of NSCs was tested in vivo and in vitro, respectively.

RESULTSNSCs displayed specific tropism for injured nerve cells or traumatic brain area in vivo and in vitro. The expression level of SDF-1 in traumatic brain area increased remarkably and the expression level of CXCR-4 in the NSCs increased simultaneously. After AMD3100 blocking the expression of CXCR-4, the migration ability of NSCs decreased significantly both in vivo and in vitro.

CONCLUSIONSSDF-1 may play a key role in stem cells migrating into injured brain area through specially combining with CXCR-4.

Animals ; Brain Injuries ; pathology ; Cell Movement ; Cells, Cultured ; Chemokine CXCL12 ; analysis ; physiology ; Neurons ; cytology ; Rats ; Receptors, CXCR4 ; analysis ; physiology ; Stem Cells ; physiology ; Tropism

BACKGROUNDA satisfactory animal model of breast cancer metastasizing to bone is unavailable. In this study, we used human breast cancer stem-like cells and human bone to build a novel "human-source" model of human breast cancer skeletal metastasis.

METHODSHuman breast cancer stem-like cells, the CD44+/CD24-/lower subpopulation, was separated and cultured. Before injection with the stem-like cells, mice were implanted with human bone in the right or left dorsal flanks. Animals in Groups A, B, and C were injected with 1 x 10(5), 1 x 10(6) human breast cancer stem-like cells, and 1 x 10(6) parental MDA-MB-231 cells, respectively. A positive control group (D) without implantation of human bone was also injected with 1 x 10(6) MDA-MB-231 cells. Immunohistochemistry was performed for determination of CD34, CD105, smooth muscle antibody, CD44, CD24, cytokine, CXC chemokine receptor-4 (CXCR4), and osteopontin (OPN). mRNA levels of CD44, CD24, CXCR4, and OPN in bone metastasis tissues were analyzed by real-time quantitative polymerase chain reaction (PCR).

RESULTSOur results demonstrated that cells in implanted human bones of group B, which received 1 x 10(6) cancer stem-like cells, stained strongly positive for CD44, CXCR4, and OPN, whereas those of other groups showed no or minimum staining. Moreover, group B had the highest incidence of human bone metastasis (77.8%, P = 0.0230) and no accompaniment of other tissue metastasis. The real-time PCR showed an increase of CD44, CXCR4, and OPN mRNA in metastatic bone tissues in group B compared with those of groups C and D, however the expression of CD24 mRNA in group B were the lowest.

CONCLUSIONSIn the novel "human source" model of breast cancer, breast cancer stem-like cells demonstrated a higher human bone-seeking ability. Its mechanism might be related to the higher expressions of CD44, CXCR4, and OPN, and the lower expression of CD24 in breast cancer stem-like cells.

Animals ; Bone Neoplasms ; secondary ; Breast Neoplasms ; pathology ; CD24 Antigen ; analysis ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Hepatocyte Growth Factor ; pharmacology ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Mice ; Neoplastic Stem Cells ; pathology ; Osteopontin ; analysis ; Phenotype ; Receptors, CXCR4 ; analysis

This study was aimed to investigate the importance of chemokine SDF-1 in maintaining proliferation ability of acute myelocytic leukemia cell line HL-60 when the effects of SDF-1 on HL-60 cell proliferation were inhibited. Marrow stromal cells were cultured and co-cultured with HL-60 cells, and SDF-1 activity was blocked with anti-CXCR4 McAb. HL-60 cell activity was detected by MTT while cell cycle and the expression of CXCR4 on HL-60 cell membrane were observed by flow cytometry meanwhile. The internal calcium ionic concentration in HL-60 cell was detected as well before and after treated with 12G5. The results showed that 12G5 down-regulated the expression of CXCR4 on HL-60 cell membrane; HL-60 cells at G(0)/G(1) phase increased, but decreased at S phase; survive rate of leukemia cells reduced; the intercellular calcium ionic concentration of HL-60 cell decreased after treated with 12G5. It was concluded that brockage of the SDF-1 activity may inhibit proliferation of leukemia cell at certain level.
Antibodies, Monoclonal ; pharmacology ; Calcium ; metabolism ; Cell Cycle ; Cell Division ; Cell Survival ; Chemokine CXCL12 ; Chemokines, CXC ; antagonists & inhibitors ; physiology ; HL-60 Cells ; cytology ; Humans ; Receptors, CXCR4 ; analysis

OBJECTIVE: To detected the mechanism of allergic rhinitis associated with asthma with bioinformatics methods. METHOD: GenMAPP software was used to analyze the expression profile of nasal mucosa of seasonal allergic rhinitis(SAR) and SAR associated with asthma of oligonucleotide microarray (Affymetrix HG-U133-plus2). One the first step,of differentially expressed genes screening were done, then differential gene database retrieval was established, at last pathway analysis was performed. RESULT: 689 genes out of 47 000 analyzed transcripts of nasal mucosa of SAR associated with asthma were differentially expressed at least 4-fold, in which 233 genes were up regulated and 456 genes were down regulated. These differential expression genes participate in 69 bio-pathways, in which the interaction pathway between cytokine and cytokine receptor was most. Chemotactic factor CXCL12 and its receptor CXCR4 expressed in SAR associated with asthma patients were up-regulated predominantly, compared with that in SAR patients. CONCLUSION Multiple pathways were involved in the development of SAR and SAR complicated with asthma. The CXCL12/CXCR4 axis might play a main role in the allergic airway diseases.
Asthma ; diagnosis ; genetics ; Chemokine CXCL12 ; metabolism ; Computational Biology ; Gene Expression Profiling ; Humans ; Oligonucleotide Array Sequence Analysis ; methods ; Receptors, CXCR4 ; metabolism ; Rhinitis, Allergic, Seasonal ; diagnosis ; genetics ; Software

OBJECTIVETo study the expression of stromal cell derived factor-1(SDF-1) in the airway and to investigate the role of SDF-1 receptor antagonist AMD3100 intervention in rats with asthma.

METHODSThirty Sprague-Dawley rats were randomly divided into three groups: normal control and asthma with and without AMD3100 intervention. The rat model of asthma was prepared by aerosolized ovalbum (OVA) challenge. The AMD3100 intervention group was administered with AMD3100 of 50 μg 30 minutes before challenge every other day, for 10 times. The characteristic airway inflammation and alterations of airway structures were observed by hemetoxylin and eosin staining. The levels of interleukin 4 and interleukin 5 in whole lung homogenates were measured using ELISA. RT-PCR was used to evaluate the expression of SDF-1 mRNA in the lung.

RESULTSThe airway wall thickness in the untreated asthma group was greater than that in the control and the AMD3100 intervention groups (P<0.05). The levels of interleukin 4 and interleukin 5 in whole lung homogenates in the AMD3100 intervention group were lower than those in the untreated asthma group (P<0.05). The expression of SDF-1 mRNA in the untreated asthma group was higher than that in the control and the AMD3100 intervention groups (P<0.05).

CONCLUSIONSSDF-1 may be associated with airway inflammation and remodeling in rats with asthma. AMD3100 may reduce the airway inflammation and improve airway remodeling by inhibiting the bioactivity of SDF-1.

Animals ; Asthma ; drug therapy ; etiology ; metabolism ; Chemokine CXCL12 ; analysis ; antagonists & inhibitors ; genetics ; physiology ; Female ; Heterocyclic Compounds ; pharmacology ; Interleukin-4 ; analysis ; Interleukin-5 ; analysis ; Lung ; metabolism ; pathology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, CXCR4 ; antagonists & inhibitors

To study the importance of chemokine SDF-1 in surviving of acute myelocytic leukemia cells HL-60, the adhesion ability of HL-60 and expression of Bcl-2, Fas protein when SDF-1 activity was blocked by anti-CXCR4 monoclonal antibody (12G5) were compared with those detected before MAb incubation, in this experiment, HL-60 cell were cultured and co-cultured with normal marrow stromal cell. The adhesion rate was detected while the expression of Bcl-2 and Fas proteins were assayed by immunohistochemical technique when SDF-1 activity was inhibited. The results showed that cell adhesion rate of HL-60 decreased while the expression Bcl-2 decreased and Fas increased. It is concluded that inhibition of SDF-1 activity increases cell apoptosis and thus reduces life-span of leukemia cell at certain level.
Antibodies, Monoclonal ; pharmacology ; Apoptosis ; Cell Adhesion ; Chemokine CXCL12 ; Chemokines, CXC ; physiology ; HL-60 Cells ; chemistry ; cytology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Receptors, CXCR4 ; physiology ; fas Receptor ; analysis

Recovery from hepatitis B virus (HBV) infection depends on the cellular immune responses. Chemokines and their receptors play significant roles in immune defense. This study was undertaken to investigate the association between HBV infection and single nucleotide polymorphisms (SNPs) of genes for the chemokines and their receptors. Between March 2002 and February 2004, a total of 957 single ethnic Korean patients were enrolled into two different groups; "HBV clearance group" (n=350), who have recovered from HBV infection, and "HBV persistence group" (n=607), who were repeatedly HBsAg-positive. The HBV persistence group was subdivided into "inactive carrier" and "HBV progression group (chronic hepatitis and cirrhosis)". We assessed polymorphisms in regulated and normal T-cell expressed and secreted (RANTES) at position -403, monocyte chemoattractant protein-1 (MCP-1) at position -2518, CCR2 V64I, CCR5 -2459, CXCR1 S276T and CXCR4 I138I using single primer extension assay. Genotype distributions of the "HBV clearance versus persistence group" and "inactive carrier versus HBV progression group" were compared. On the basis of unconditional logistic regression analysis with adjustment for age and sex, no statistically significant association with susceptibility to persistent HBV infection was observed with RANTES -403, MCP-1 -2518, CCR2 V64I, CCR5 -2459, CXCR1 S276T, and CXCR4 I138I polymorphisms. In addition, no association of analyzed SNPs with HBV disease progression was found.
Chemokine CCL2/*genetics ; Chemokine CCL5/*genetics ; Disease Progression ; Genotype ; Hepatitis B/ethnology/*genetics/*therapy ; Hepatitis B virus/metabolism ; Humans ; Korea ; *Polymorphism, Genetic ; Receptors, CCR2 ; Receptors, CCR5/*genetics ; Receptors, CXCR4/*genetics ; Receptors, Chemokine/*genetics ; Receptors, Interleukin-8A/*genetics ; Regression Analysis ; Treatment Outcome