A functional interaction between the CCR5 and CD34 molecules expressed in hematopoietic cells can support (or even promote) the development of cancer

Abstract Inflammation and angiogenesis are linked to the development of cancer since both can support the establishment of a tumor-prone microenvironment. The CCR5 is a major regulatory molecule involved in inflammation. The CD34 molecule is commonly described as a hematopoietic stem cell marker, and CD34+ cells are involved in the regulation of distinct physiological processes, including angiogenesis. CCR5 participates in the development of various types of cancer, and recently, a reduced CCR5 expression was associated with low CD34+ cell counts in human cord blood. A naturally occurring genetic variant of the CCR5 gene, the so-called CCR5Δ32 polymorphism, consists of a 32 base-pair deletion in the DNA, interfering in the CCR5 protein levels on the cell surface. When in homozygosis, this variant leads to a total absence of CCR5 expression on the cell surface. In heterozygous individuals, CCR5 surface levels are reduced. Based on these key findings, we hypothesize that a functional interaction can connect CCR5 and CD34 molecules (giving rise to a "CCR5-CD34 axis"). According to this, a CCR5-CD34 interaction can potentially support the development of different types of cancer. Consequently, the lack of CCR5 in association with reduced CD34+ cell counts could indicate a protective factor against the development of cancer. It is required to characterize in detail the functional relationship between CCR5 and CD34 proteins, as well as the real influence of both molecules on the susceptibility and development of cancer at population level. If our hypothesis is confirmed, the CCR5-CD34 axis may be a potential target in the development of anti-cancer therapies.

Small molecule CCR5 antagonists in clinical trials and their drug resistances:research advances / 国际药学研究杂志

C-C chemokine receptor 5 (CCR5), one of the major co-receptors of HIV-1, can mediate the fusion of HIV-1 to cell membranes. CCR5 antagonists can bind to CCR5 and cause conformational changes in CCR5, thus blocking HIV-1 infection. Several small molecule CCR5 antagonists with strong activity and good tolerance have been screened and entered the clinical trials. With the widespread use of CCR5 antagonists, drug resistance has gradually emerged. There are many reports about of drug-related failure in vivo and in vitro. Therefore, this review summarizes the mechanism of CCR5-mediated HIV-1 infection, the research progress in maraviroc and other CCR5 antagonists which have entered clinical trials and their drug resistance.

Effects of antagonistic peptides binding specifically with first and second extracellular loops of CCR5 on colitis rats induced by TNBS / 中国病理生理杂志

AIM:To study the effects of antagonistic peptides binding specifically with the first and second extracellular loops (ECL1 and ECL2) of C-C chemokine receptor 5 (CCR5) on the colitis rats induced by trinitrobenzenesulfonic acid (TNBS) and the mechanisms.METHODS:The colitis model of SD rats was induced by TNBS (100 mg/kg).The effects of 2 antagonistic peptides at different doses (ECL1:25, 35 and 45 mg/kg;ECL2:15, 25 and 35 mg/kg) on the model rats including the changes of disease activity index (DAI), colon macroscopic damage index (CMDI) and histological grading were observed.The mRNA and protein expression levels of TNF-α and COX-2 in the colonic mucosa were detected by real-time PCR and Western blot, respectively.RESULTS:Compared with model group, the changes of DAI, CMDI and histopathological injury of the rats treated with ECL2 antagonistic peptide HY at an appropriate dose were significantly reduced (P<0.05), and the protein and mRNA expression levels of TNF-α and COX-2 were significantly decreased (P<0.05).However, the effects of ECL1 antagonistic peptide GH on all scores and the expression levels of TNF-α and COX-2 were not obvious.CONCLUSION:ECL2 antagonistic peptide HY relieves TNBS-induced colitis in SD rats via down-regulating the expressions of TNF-α and COX-2 in the colonic mucosa, while the effect of ECL1 antagonist peptide GH was not obvious.

Therapeutic effect of folic acid combined with vitamin B12 on atherosclerosis induced by hyperhomocysteinemia in rats / 吉林大学学报(医学版)

Objective:To investigate the therapeutic effect of folic acid combined with vitamin B12 on atherosclerosis induced by hyperhomocysteinemia in the rats,and to elucidate its therapeutic mechanism.Methods:Thirty-three rats were randomly divided into control group,model group and treatment group (n=11).The rats in control group were given normal diet,the rats in model group were given normal diet and methionine,and the rats in treatment group were given normal diet,methionine,perfused folic acid and vitamin B12.The histopathological changes of thoracic aorta of the rats were observed by HE staining.The levels of serum Hcy and matrix metalloproteinase-9 (MMP-9) were detected with ELISA,and the expression levels of CCR5 and CC chemokine ligand 5 (CCL5) proteins in thoracic aortic smooth muscle cells were detected by immunohistochemical staining.The expression levels of CCR5 and CCL5 mRNA in thoracic aortic smooth muscle cells were detected by RT-PCR.Results:The results of HE staining showed that the medial smooth muscle cells of the rat thoracic aorta were arranged neatly in control group,the intimal elastic membrane was complete and the endothelial cells were continuous;the smooth muscle cells of the thoracic aorta were loose in model group,the endometrium was damaged,the foam cells were formed under the intima,and the endothelial cells were necrotic;the pathological changes of the thoracic aorta of the rats in treatment group were between control group and model group.The levels of serum Hcy of the rats in model group and treatment group were higher than that in control group (P＜0.05);the serum Hcy level in treatment group was lower than that in model group (P＜0.05).The levels of serum MMP-9 of the rats in model group and treatment group were higher than that in control group (P＜0.05);the serum MMP-9 level in treatment group was lower than that in model group (P＜0.05).The expression gray values of CCR5 and CCL5 proteins in the thoracic aortic smooth muscle cells of the rats in rats in model group and treatment group were lower than those in control group (P＜0.05);the expression gray values of CCR5 and CCL5 proteins in the thoracic aortic smooth muscle cells of the rats in treatment group were higher than those in model group (P＜0.05).The relative expression levels of CCR5 and CCL5 mRNA in the thoracic aortic smooth muscle cells of the rats in model group and treatment group were higher than those in control group (P＜0.05);the relative expression levels of CCR5 and CCL5 mRNA in the thoracic aortic smooth muscle cells of the rats in treatment group was lower than that in model group (P＜0.05).Conclusion:Folic acid combined with vitamin B12 could decrease the serum Hcy and MMP-9 levels and the expression levels of CCR5 and CCL5 in the thoracic aortic smooth muscle cells which may be one of the mechanisms of folic acid combined with vitamin B12 in the treatment of atherosclerosis.

Therapeutic effect of folic acid combined with vitamin B12 on atherosclerosis induced by hyperhomocysteinemia in rats / 吉林大学学报(医学版)

Objective:To investigate the therapeutic effect of folic acid combined with vitamin B12 on atherosclerosis induced by hyperhomocysteinemia in the rats,and to elucidate its therapeutic mechanism.Methods:Thirty-three rats were randomly divided into control group,model group and treatment group (n=11).The rats in control group were given normal diet,the rats in model group were given normal diet and methionine,and the rats in treatment group were given normal diet,methionine,perfused folic acid and vitamin B12.The histopathological changes of thoracic aorta of the rats were observed by HE staining.The levels of serum Hcy and matrix metalloproteinase-9 (MMP-9) were detected with ELISA,and the expression levels of CCR5 and CC chemokine ligand 5 (CCL5) proteins in thoracic aortic smooth muscle cells were detected by immunohistochemical staining.The expression levels of CCR5 and CCL5 mRNA in thoracic aortic smooth muscle cells were detected by RT-PCR.Results:The results of HE staining showed that the medial smooth muscle cells of the rat thoracic aorta were arranged neatly in control group,the intimal elastic membrane was complete and the endothelial cells were continuous;the smooth muscle cells of the thoracic aorta were loose in model group,the endometrium was damaged,the foam cells were formed under the intima,and the endothelial cells were necrotic;the pathological changes of the thoracic aorta of the rats in treatment group were between control group and model group.The levels of serum Hcy of the rats in model group and treatment group were higher than that in control group (P＜0.05);the serum Hcy level in treatment group was lower than that in model group (P＜0.05).The levels of serum MMP-9 of the rats in model group and treatment group were higher than that in control group (P＜0.05);the serum MMP-9 level in treatment group was lower than that in model group (P＜0.05).The expression gray values of CCR5 and CCL5 proteins in the thoracic aortic smooth muscle cells of the rats in rats in model group and treatment group were lower than those in control group (P＜0.05);the expression gray values of CCR5 and CCL5 proteins in the thoracic aortic smooth muscle cells of the rats in treatment group were higher than those in model group (P＜0.05).The relative expression levels of CCR5 and CCL5 mRNA in the thoracic aortic smooth muscle cells of the rats in model group and treatment group were higher than those in control group (P＜0.05);the relative expression levels of CCR5 and CCL5 mRNA in the thoracic aortic smooth muscle cells of the rats in treatment group was lower than that in model group (P＜0.05).Conclusion:Folic acid combined with vitamin B12 could decrease the serum Hcy and MMP-9 levels and the expression levels of CCR5 and CCL5 in the thoracic aortic smooth muscle cells which may be one of the mechanisms of folic acid combined with vitamin B12 in the treatment of atherosclerosis.

Effects of antagonistic peptide specifically binding to second extracellular loop of CCR5 on inflammatory cell infiltration and TNF-α expression in lung tissues of asthmatic mice induced by OVA / 中国病理生理杂志

AIM: To investigate the effects of antagonistic peptide specifically binding to the second extracellular loop of CC chemokine receptor 5 (CCR5) on inflammatory cell infiltration and TNF-α expression in lung tissues of asthmatic mice.METHODS: The asthmatic model of BALB/c mice was induced by ovalbumin (OVA) and the optimal sensitization concentration of OVA was screened.After modeling, the mice were intervened by gradual concentrations of antagonistic peptide via tail-vein injection.The pathocytological analysis and grading were performed in the lung tissues with HE staining.The expression of TNF-α at mRNA and protein levels in the lung tissues was determined by real-time PCR and Western blot.RESULTS: The optimal concentration of OVA was 500 mg/L (0.1 mL) as this concentration of OVA stably induced moderate degree of inflammation in the BALB/c mice.Treatment with different concentrations (1.5 g/L, 2.5 g/L and 3.5 g/L) of antagonistic peptide at 0.2 mL through tail-vein injection inhibited the expression of TNF-α, and markedly reduced the degree of inflammation in the lung tissues.The optimal concentration of antagonistic peptide was 2.5 g/L as the lung inflammation degree in 2.5 g/L group alleviated by 2 grades, and the number of inflammatory cells was also significantly reduced.Moreover, the mRNA expression abundance of TNF-α was nearly decreased by 90%, and the protein expression of TNF-α was decreased by 70% compared with model group.Meanwhile, the use of antagonistic peptide at 2.5 g/L before OVA stimulation confirmed the preventive function to some degree.In this group, the lung inflammation degree alleviated by 1 grade, and the expression of TNF-α at both mRNA and protein levels decreased by nearly 50%.CONCLUSION: The antagonistic peptide of CCR5 effectively inhibits the expression of TNF-α and relieves the inflammation in the asthmatic mouse lung tissues in a concentration-dependent manner.

Panning and identification of antagonistic active peptides specifically bin-ding to the first and second extracellular membrane loops of rat CCR5 by technique of phage display peptide library / 中国病理生理杂志

[ ABSTRACT] AIM: To pan the active peptides which specifically bound to the first and second extracellular membrane loops of rat CC chemokine receptor 5 ( CCR5 ) .METHODS: The technique of phage display peptide library was used and binding ability of the peptides was identified.The amino acid sequences of the first and second extracellular loops of rat CCR5 were searched in the protein database and chemically synthesized corresponding linear peptides were used as targets in the biopanning.After 3 to 4 rounds of screening with Ph.D.TM-7 Phage Display Peptide Library were per-formed, the specific phages were collected and primarily identified by ELISA.RESULTS:The sequences of the peptides displayed on the selected phages were GHWKVWL and HYIDFRW, both of them exhibited positive in phage binding ELISA and the binding to phages and targets were concentration dependent and saturable.CONCLUSION:Two antagonis-tic active peptides specifically binding to CCR5 were successfully obtained by the technique of phage display peptide librar-y, and the binding ability to the first and second extracellular membrane loops of rat CCR5 were proved in vitro.

Toll like receptor 2 and CC chemokine receptor 5 cluster in the lipid raft enhances the susceptibility of Leishmania donovani infection in macrophages.

In experimental visceral leishmaniasis the causative obligate protozoan parasite, L. donovani invades and multiplies inside of macrophages, one of the sentries of the mammalian immune system. The initial host-parasite interaction between the Leishmania promastigote and the macrophage takes place at the plasma membrane interface. To trace any possible interaction between Toll-like receptor 2 (TLR2) and CC chemokine receptor 5 (CCR5) during early Leishmania-macrophage interactions, it was observed that the expression of both TLR2 and CCR5 were significantly increased, along with their recruitment to the lipid raft. TLR2 silencing attenuates CCR5 expression and restricts L. donovani infection, indicating a regulatory role of TLR2 and CCR5 during infection. Silencing of CCR5 and TLR2 markedly reduced the number of intracellular parasites in macrophages by host protective cytokine responses, while raft disruption using β-MCD affected TLR2/CCR5 cross-talk and resulted in a significant reduction in parasite invasion. In vivo RNA interference of TLR2 and CCR5 using shRNA plasmids rendered protection in Leishmania donovani-infected mice. Thus, this study for the first time demonstrates the importance of TLR2/CCR5 crosstalk as a significant determinant of Leishmania donovani entry in host macrophages.

Distribution of CC-chemokine receptor-5-32 allele among the tribal and caste population of Vidarbha region of Maharashtra state.

BACKGROUND: Genetic relationships among the ethnic groups are not uniform across the geographical region. Considering this assumption, we analyzed the frequency of the CC-chemokine receptor-5 (CCR5)-32 allele of the CCR5 chemokine receptor, which is considered a Caucasian marker, in Bhil tribal and Brahmin caste sample sets from the population. MATERIALS AND METHODS: 108 blood samples were collected from 6 tribe's populations and a caste population from the district of Vidarbha region. RESULTS AND DISCUSSION: The presence of low frequencies of CCR5-Δ32 in an individual of Bhil tribe (0.034, χ2 value 0.017) in the present study implies that these communities may have a better resistance toward human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) than the other studied tribe sample, as non-show such mutation. CONCLUSION: The marginal presence of the allele seen in the studied tribal population could be due to gene flow from the people of European descent. However, lack of the homozygous CCR5-Δ32 mutation and the low prevalence of heterozygous CCR5-Δ32 mutations suggest that the Indians are highly susceptible to HIV/AIDS, and this correlates with the highest number of HIV/AIDS infected individuals in India.

Chemokine Systems Link Obesity to Insulin Resistance

Obesity is a state of chronic low-grade systemic inflammation. This chronic inflammation is deeply involved in insulin resistance, which is the underlying condition of type 2 diabetes and metabolic syndrome. A significant advance in our understanding of obesity-associated inflammation and insulin resistance has been recognition of the critical role of adipose tissue macrophages (ATMs). Chemokines are small proteins that direct the trafficking of immune cells to sites of inflammation. In addition, chemokines activate the production and secretion of inflammatory cytokines through specific G protein-coupled receptors. ATM accumulation through C-C motif chemokine receptor 2 and its ligand monocyte chemoattractant protein-1 is considered pivotal in the development of insulin resistance. However, chemokine systems appear to exhibit a high degree of functional redundancy. Currently, more than 50 chemokines and 18 chemokine receptors exhibiting various physiological and pathological properties have been discovered. Therefore, additional, unidentified chemokine/chemokine receptor pathways that may play significant roles in ATM recruitment and insulin sensitivity remain to be fully identified. This review focuses on some of the latest findings on chemokine systems linking obesity to inflammation and subsequent development of insulin resistance.

Chemokine Receptor 5 of Human Brain Microvascular Endothelial Cells Takes Part in Amyloid beta Induced T Lymphocytes Transendothelial Migration / 中国医科大学学报

Objective To investigate the function of chemokine receptor 5（CCR5）in human brain microvascular endothelial cells（HBMEC）during T lymphocytes transendothelial migration induced by amyloid beta（Aβ）.Methods The in vitro model of blood-brain barrier was established with cultured HBMEC monolayer.The ability of T lymphocytes transendothelial migration induced by Aβ was analyzed by Transwell and Millicell-ERS endothelia volt-ohmmeter.Results Aβ could promote T lymphocytes to migrate through the HBMEC monolayer.Over-expression of CCR5 in HBMEC promoted T lymphocytes to migrate through HBMEC monolayer.Expression of CCR5 mutant in HBMEC inhibited the transendothelial migration of T lymphocytes.Conclusion CCR5 of HBMEC is involved in T lymphocytes transendothelial migration induced by Aβ.

Expression of chemokine receptor CCR5 in human follicular thyroid carcinoma / 第二军医大学学报

Objective: To study the expression of chemokine receptor CCR5 in follicular thyroid carcinoma and the serum level of CCR5 ligand, so as to assess the role of CCR5 in progression and metastasis of follicular thyroid carcinoma. Methods: Fifteen samples of follicular thyroid carcinoma, 17 samples of follicular thyroid adenoma and 12 adjacent normal samples were analyzed immunohistochemically for CCR5 expression. The sera concentrations of CCL3, CCL4 and CCL5 were measured by ELISA in all patients. Results: CCR5 was positive in follicular thyroid carcinoma samples, with the positive rate being 73.33%, and was not detected in the follicular thyroid adenoma and the normal samples (P<0.01). The concentrations of CCL3 and CCL5 in the sera of follicular thyroid carcinoma patients were significantly higher than those of the other 2 groups (P<0.05). Conclusion: CCR5 is highly expressed in follicular thyroid carcinoma tissues and the concentrations of CCL3 and CCL5 are obviously increased in the sera of patients, indicating that CCR5 may play an important role in the pathogenesis of follicular thyroid carcinoma.

The association of chemokine receptor 5 gene 59029A/G polymorphisms with diabetic nephropathy in type 2 diabetic patients / 대한내과학회지

BACKGROUND/AIMS: Chemokine receptor 5 (CCR5) is a receptor for several chemokines, including regulated upon activation, normal T-cell-expressed and -secreted (RANTES; also known as CCL5) and macrophage inflammatory protein 1 (MIP-1), and mediates the recruitment of monocytes and their differentiation to macrophages during the inflammatory process. As such, CCR5 plays an important role in the development and progression of atherosclerosis, which has an underlying inflammation component and contributes to the development or progression of diabetic complications. Several studies have reported that a genetic variation of the CCR5 gene with A/G at basepair 59029 (59029 A/G) was associated with diabetic complications, although conflicting data exist. We evaluated the association of the CCR5 59029 A/G polymorphism with diabetic complications in type 2 diabetes patients. METHODS: We conducted a case-control study, enrolling 325 patients with type 2 diabetes. We examined the association of CCR5 genotypic variations with diabetic nephropathy, retinopathy, and macrovascular complications such as coronary heart disease and stroke. Genotyping was performed using the polymerase chain reaction and restriction fragment length polymorphism technology with Bsp1286I restriction enzyme. RESULTS: We determined no allelic association of CCR5 59029 A/G with diabetic nephropathy (p=0.500) in the male or female patients. Diabetic retinopathy and macrovascular complications such as coronary heart disease and stroke were not associated with the 59029 A/G polymorphism. Among those patients with diabetic nephropathies, those with the GG genotype showed a tendency toward higher serum levels of LDL-cholesterol. CONCLUSIONS: These results suggest that that the 59029 A/G polymorphism of the CCR5 gene is not associated with diabetic complications in type 2 diabetes patients. Further studies are required to understand the role of CCR5 polymorphisms in the development of diabetic complications.

Association between CCR5 Promoter Polymorphisms and Hepatitis B Virus Infection / 대한간학회지

BACKGROUND/AIMS: Immunogenetic factors may play a role in determining the susceptibility of an individual to viral infection. CCR5 promoter polymorphisms are known to be associated with HIV infection. However, there has been no report on the association between CCR5 promoter polymorphism and HBV infection. Therefore, we investigated the relationship between the CCR5 promoter polymorphism and HBV infection. METHODS: A total of 377 patients were classified into two groups according to their HBV infection status: (1)he spontaneous clearance group (SC); HBsAg (-), anti-HBc (+), anti-HBs (+) (2)he chronic HBsAg (+) carrier group (CC); HBsAg (+), anti-HBc (+), anti-HBs (-). CCR5 polymorphisms were detected by employing matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS)- based SNP scoring assay, termed Restriction Fragment Mass Polymorphism (RFMP), which exploits the differences in molecular masses between the common allele and rare allele bases of interest. RESULTS: We found that the genotype frequencies of CCR5 A59029G significantly differed between the SC group (n=138) and CC group (n=239) (P<0.05). The CCR5 59029A allelic genotype was associated with an increased risks of chronic infection rather than spontaneous clearance (P=0.002), and the presence of the CCR5 59029G allele was significantly associated with the spontaneous clearance of HBV (P=0.001). Strong linkage disequilibrium between the CCR5-59029 and the CCR5-59353 polymorphic variants was identified. None of the 377 subjects had the CCR5-32 bp deletion mutation. CONCLUSIONS: The CCR5 promoter polymorphisms at position 59029 might play a role in the clearance of HBV infection. This primary experimental evidence needs further studies to clarify the clinical usefulness of CCR5 promoter polymorphisms as a target for the screening or treatment of HBV infection.

Expression of chemokine receptor CCR5 in human follicular thyroid carcinoma / 第二军医大学学报

Objective:To study the expression of chemokine receptor CCR5 in follicular thyroid carcinoma and the serum level of CCR5 ligand,so as to assess the role of CCR5 in progression and metastasis of follicular thyroid carcinoma.Methods:Fifteen samples of follicular thyroid carcinoma,17 samples of follicular thyroid adenoma and 12 adjacent normal samples were analyzed immunohistochemically for CCR5 expression.The sera concentrations of CCL3,CCL4 and CCL5 were measured by ELISA in all patients.Results:CCR5 was positive in follicular thyroid carcinoma samples,with the positive rate being 73.33%,and was not detected in the follicular thyroid adenoma and the normal samples(P

Effects of anti-chemokine receptor-5 antibody on type 1 diabetes / 第二军医大学学报

Objective:To explore the anti-diabetogenic effect of antibody blocking portion of chemokine receptor-5 antibody(CCR5) molecule.Methods: The severe combined immunodeficient NOD(NOD.Scid) mice,injected i.p.with splenocytes from nonobese diabetic mouse,were randomly divided into 2 groups(8 of each group): anti-CCR5 Ab group and PBS group.The mice in anti-CCR5 Ab group were treated with a specific polyclonal antibody,targeting the first extracellular loop of CCR5,and PBS group was treated with PBS.Blood glucose levels were measured to observe the anti-diabetogenic effect of the antibody.Histological examination and ELISA analysis were also performed.Results: Within 70 d,all of the control mice developed diabetes while 5 of the 8 mice treated with anti-CCR5 Ab were diabetes-free.The difference of inflammation score between 2 groups was significant(P

Blocking the CC chemokine receptor 5 pathway by antisense peptide nucleic acid prolongs islet allograft survival / 中国普通外科杂志

Objective To identify the effect of peptide nucleic acid of CC chemokine receptor 5 on acute rejection of islet allograft.Methods Mice islet transplant models were used to test the effect of PNA CCR5 by targeting CCR5 in acute allograft rejection.In vitro T cell proliferative responses were assessed by mixed lymphocyte response(MLR).RT-PCR and Western blot were used to detect the expression of mRNA and protein.Results PNA CCR5-treated recipients demonstrated statistically significant prolongation(12.00?1.75)d in functional allograft survival when compared with saline(6.50?0.58)d or PNA mismatch-treated recipients(6.50?0.50)d.The CCR5 mRNA expression level of PNA CCR5,control,and PNA mismatch treatment recipients at day 7 posttransplant was 0.56?0.05,1.68?0.07 and 1.80?0.14,respectively.The data showed that CCR5 protein was significantly down-regulated in PNA CCR5 treatment allografts compared with saline and PNA mismatch treatment allografts(P

The preparation of first extracellular membrane loop1 of human monocyte chemotactic protein receptor 5 N- terminal's specific antibody F(ab′) 2 / 中国免疫学杂志

Objective:To prepare the first extracellular domain of human ? chemokine receptor 5(huCCR5)NH 2 terminal's specific antibody F(ab′) 2 and its detection method Methods:Use computer analyzed and located the least homologous domain of the extracellular first loops The gene fragment was amplified by PCR and cloned into the pGEX IN vector A recombinant GST fusion protein was constructed After comfirming the correctness of the inserted sequence,the transformation and expression of this fusion protein were performed in E coli The expression products of the fusion protein were purified and 2 New Zealand rabbits were immunized An anti huCCR5 NH 2 terminal antibody F(ab′) 2 was prepared by protein A Sepharose CL 4B affinity chromatography ,pepsin digestion and S 200 column seperation.Results:Reduced,unreduced SDS PAGE and ELISA block examination analysis demonstrated that this F(ab′) 2 had high specificity to combine with huCCR5 Conclusion:In this paper,not only introduce a simple and quick method to get a specific antibody F(ab′) 2 of certain functional domain but also a good idea and technique to study other high similar superfamily members

Expression of chemokine receptor CCR5 in human invasive ductal breast carcinoma / 第二军医大学学报

Objective:To detect the expression of chemokine receptor CCR5 in human invasive ductal breast carcinoma and explore the role of CCR5 in breast cancer.Methods: Thirty samples of invasive ductal breast carcinoma and 10 samples of breast fibroadenoma were analyzed separately by immunohistochemical staining for CCR5 expression. MIP1?,MIP1? and RANTES concentrations were measured by ELISA in the sera of patients. Results: The expression of CCR5 was detected in invasive ductal breast carcinoma (the positive rate was 53.3%) and was not detected in breast fibroadenoma. RANTES concentrations in the serum of breast carcinoma patients increased significantly (P