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Objective@#To investigate whether poly (lactic-co-glycolic acid) (PLGA) as protein delivery vehicles that encapsulate CC chemokine receptor 5 antibody (anti-CCR5) has more suppressive function on macrophages than single anti-CCR5 in mouse endometriosis model.@*Methods@#The PLGA/anti-CCR5 nanoparticles were synthesized. The cumulative release of anti-CCR5 from PLGA/anti-CCR5 nanoparticles was evaluated. The mouse endometriosis model was established and divided into control group, anti-CCR5 group and PLGA/anti-CCR5 group. Meanwhile, ectopic endometrial cells (EEC) and macrophages isolated from peritoneal fluid were cultured in vitro. Flow cytometry was used to detect the proportion of macrophages in the peritoneal fluid of each group. The secretion of interleukin 10 (IL-10) and transforming growth factor β (TGF-β) in each group were determined by ELISA. The proliferation and infiltration of EEC were detected by 5-bromodeoxyuridine proliferation kit and matrigel invasion kit.@*Results@#The PLGA/anti-CCR5 nanoparticles were successfully synthesized. The mouse endometriosis model was established and the EEC and macrophages were cultured. Compared with the anti-CCR5 without nanoparticles, the bioconjugate PLGA/anti-CCR5 nanoparticles could control the release of anti-CCR5 from day 3 to day 24. The proportion of macrophages in PLGA/anti-CCR5 group were gradually reduced compared with those in anti-CCR5 group (P<0.01), the ratios of day 7 [(4.5±1.5)%] and day 3 [(6.3±0.6)%], day 14 [(2.6±0.7)%] and day 7 were significantly different (P<0.01 and P<0.05). PLGA/anti-CCR5 reduced IL-10 and TGF-β levels relative to anti-CCR5 (P<0.01),and decreased gradually on day 3, day 7, and day 14 (P<0.01). Anti-IL-10+anti-TGF-β could reduce the proliferation [(70.8±7.6)%] and invasion ability [(50.2±9.1)%] of EEC (P<0.05).@*Conclusions@#In mouse endometriosis model, PLGA/anti-CCR5 may inhibit the proliferation and invasion of EEC by inhibiting the secretion of IL-10 and TGF-β by macrophages, suggesting that it provide a new idea for the treatment of clinical endometriosis.
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Objective To investigate whether poly (lactic?co?glycolic acid) (PLGA) as protein delivery vehicles that encapsulate CC chemokine receptor 5 antibody (anti?CCR5) has more suppressive function on macrophages than single anti?CCR5 in mouse endometriosis model. Methods The PLGA/anti?CCR5 nanoparticles were synthesized. The cumulative release of anti?CCR5 from PLGA/anti?CCR5 nanoparticles was evaluated. The mouse endometriosis model was established and divided into control group, anti?CCR5 group and PLGA/anti?CCR5 group. Meanwhile, ectopic endometrial cells (EEC) and macrophages isolated from peritoneal fluid were cultured in vitro. Flow cytometry was used to detect the proportion of macrophages in the peritoneal fluid of each group. The secretion of interleukin 10 (IL?10) and transforming growth factor β (TGF?β) in each group were determined by ELISA. The proliferation and infiltration of EEC were detected by 5?bromodeoxyuridine proliferation kit and matrigel invasion kit. Results The PLGA/anti?CCR5 nanoparticles were successfully synthesized. The mouse endometriosis model was established and the EEC and macrophages were cultured. Compared with the anti?CCR5 without nanoparticles, the bioconjugate PLGA/anti?CCR5 nanoparticles could control the release of anti?CCR5 from day 3 to day 24. The proportion of macrophages in PLGA/anti?CCR5 group were gradually reduced compared with those in anti?CCR5 group (P<0.01), the ratios of day 7 [(4.5±1.5)%] and day 3 [(6.3±0.6)%], day 14 [(2.6±0.7)%] and day 7 were significantly different (P<0.01 and P<0.05). PLGA/anti?CCR5 reduced IL?10 and TGF?β levels relative to anti?CCR5 (P<0.01),and decreased gradually on day 3, day 7, and day 14 (P<0.01). Anti?IL?10+anti?TGF?β could reduce the proliferation [(70.8 ± 7.6)% ] and invasion ability [(50.2 ± 9.1)% ] of EEC (P<0.05). Conclusions In mouse endometriosis model, PLGA/anti?CCR5 may inhibit the proliferation and invasion of EEC by inhibiting the secretion of IL?10 and TGF?β by macrophages, suggesting that it provide a new idea for the treatment of clinical endometriosis.
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Objective To evaluate the role of spinal C-C motif chemokine receptor 5 (CCRS) in remifentanil-induced hyperalgesia in rats with incisional pain.Methods Thirty-two male Sprague-Dawley rats,weighing 240-260 g,aged 2-3 months,in which intrathecal and caudal catheters were successfully implanted,were divided into 4 groups (n =8 each) using a random number table:control group (group C),CCR5 antagonist maraviroc group (group M),remifentanil plus incisional pain group (group R + I) and maraviroc plus remifentanil plus incisional pain group (group M + R + I).Phosphate buffer solution (PBS) 10 μl was intrathecally injected and normal saline was infused for 60 rnin at 1 μg · kg-1 · min-1 via the caudal vein in group C.Maraviroc 100 pmol (in 10 μl of PBS) was intrathecally injected and normal saline was infused for 60 min at 1 μg · kg-1 · min-1 via the caudal vein in group M.PBS 10 μl was intrathecally injected,then the model of incisional pain was established,and remifentanil 1 μg · kg-1 · min-1 was infused for 60 min via the caudal vein in group R+I.Maraviroc 100 pmol (in 10 μl of PBS) was intrathecally injected,then the model of incisional pain was established,and remifentanil 1 μg · kg-1 · min-1 was infused for 60 min via the caudal vein in group M+R+I.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 24 h before infusion of remifentanil or normal saline (T0) and 2,6,24 and 48 h after stopping infusion (T1-4).The rats were sacrificed after the last measurement of pain threshold,and L4-6 segments of the spinal cord were removed for determination of the expression of glial fibrillary acidic protein (GFAP) and ionized calciumbinding adapter molecule-1 (Iba-1) by Western blot.Results Compared with group C,the MWT was significantly decreased and TWL was shortened at T1-4,and the expression of GFAP and Iba-1 in the spinal cord was up-regulated in R+I and M+R+I groups (P＜0.05),and no significant change was found in the parameters mentioned above in group M (P＞0.05).Compared with group R+I,the MWT was significantly increased and TWL was prolonged at T1-4,and the expression of GFAP and Iba-1 in the spinal cord was down-regulated in group M+R+I (P＜0.05).Conclusion Spinal CCR5 is involved in remifentanil-induced hyperalgesia in the rats with incisional pain,and the mechanism may be related to activating astrocytes and microglias.
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Background Acquired immune deficiency syndrome (AIDS) is an infectious disease caused by human immunodeficiency virus (HIV).Highly active antiretroviral therapy (HAART) is an effective treatment for AIDS,but it cannot completely eliminate the viral load in the body for the existence of HIV reservoir.Previous studies demonstrated that HIV could be detected in tears of virus load negative AIDS patients who received effective HAART,suggesting that lacrimal gland is another member of HIV reservoirs.Objective The aim of this study was to explore whether lacrimal gland has a molecular basis of HIV infection and the mechanism of lacrimal gland infection of HIV.Methods Fourteen specimens of lacrimal gland were collected during the surgery from 14 patients with lacrimal gland diseases in Peking Union Medical College Hospital from November 2013 to December 2015,including 13 non-HIV-infected patients and 1 HIV-infected patient.In 13 non-HIV infected patients,lacrimal glands prolapse was in 12 patients with the normal pathological tissue structure and dacryoadenitis was in 1 patient with the histopathological diagnosis of interstitial lymphoid tissue hyperplasia.The clinical manifestation of HIV-infected patient was dacryoadenitis with the histopathological diagnosis of interstitial lymphoid tissue hyperplasia.The paraffin sections of 12 non-HIV-infected specimens and 1 HIV-infected specimen were prepared,and the expressions of CD4,C-X-C chemokine receptor 4 (CXCR4) and C-C chemokine receptor type 5 (CCR5) in lacrimal gland specimens were detected by immunohistochemistry and verified in 1 specimen of non-HIV-infected specimen by immunofluorescence technology.Results Immunohistochemistry showed that CD4 was suspiciously positive expression in non-HIV-infected specimens with the strong background staining.CXCR4 was positively expressed in cytoplasm and nuclei of most lacrimal epithelial cells of lacrimal gland epithelial cells in each specimen,and CCR5 was focally expressed in few lacrimal gland epithelial cells in each specimen.In addition,CD4,CXCR4 and CCR5 were positively expressed in intercellular scattered lymphocytes on the specimens.Immunofluorescence assay showed that CD4,CXCR4 and CCR5 were expressed in the specimens with the red fluorescence,with the linear-and patchy-like distribution mainly in cellular membrane for CD4 or spot-like distribution for CXCR4 and CCR5 in the cytoplasm.Conclusions HIV receptor CD4 and accessory receptor CXCR4,CCR5 are positively expressed in the lacrimal gland epithelial cells,which is the molecular basis of HIV infection and become a potential HIV reservoir preventing HIV eradication.
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Objective To evaluate the changes in the expression of CC-chemokine ligand 3 (CCL3) and CC-chemokine receptor 5 (CCR5) in the spinal cord during hyperalgesia induced by remifentanil in rats with incisional pain.Methods Thirty-two male Sprague-Dawley rats,aged 2-3 months,weighing 240-260 g,were randomly divided into 4 groups (n=8 each) using a random number table:control group (group C),incisional pain group (group Ⅰ),remifentanil group (group R) and remifentanil+incisional pain group (group R+I).A 1-cm longitudinal incision was made in the plantar surface of the left hindpaw in anesthetized rats.While the model of incisional pain was established,remifentanil was infused for 60 min at 1 μg · kg-1 · min-1.At 24 h before infusion of remifentanil (baseline) and 2,6,24 and 48 h after the end of infusion,the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.The rats were sacrificed after the last measurement of pain threshold,the lumbar segment (L4-6) of the spinal cord was removed for determination of CL3 and CCR5 mRNA expression (by real-time PCR) and CL3 and CCR5 expression (by Western blot).Results Compared with group C,the MWT was significantly decreased,the TWL was shortened,and the expression of CCL3 and CCR5 mRNA and protein was up-regulated in I,R and R+ I groups.Compared with I and R groups,the MWT was significantly dccreascd,the TWL was shortened,and the expression of CCL3 and CCR5 mRNA and protein was up-regulated in group R+I.Conclusion The mechanism by which remifentanil induces hyperalgesia is related to up-regulated expression of CCL3 and CCR5 in the spinal cord of rats with incisional pain.
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Objective To study biological characteristics of R5 tropic human immunodeficiency virus (HIV)-1 strains in different disease stage. Methods Primary clinical viruses were isolated from fresh peripheral blood mononuclear cells (PBMC) using co-culture methods; meanwhile, viral co receptor usage and infectivity were tested using flow cytometry on GHOST (3) cell lines,which expressed CD4 receptor and CC ehemokine receptor 5 (CCR5) or CXCR4 eoreceptor; to identified CCR5 tropic viruses(R5 tropic strains). Viral replication kinetics was detected in PBMCs. Plasma viral load was measured using an HIV-1 nucleotide fluorescence quantification assay kit. Results There were 22 individuals with HIV-1 subtype B' infection, in which 11 were CD4>0. 2 × 109/L and 11 were CD4≤0. 2 × 109/L. All isolated viruses used CCR5 coreceptor and therefore were HIV-1 R5 tropic strains. The infectivity of R5 tropic strains isolated from patients with CD4≤0.2 × 109/L was (7.392 7 ± 4. 584 2) % ; while the infectivity of R5 tropic strain from patients with CD4>0. 2 × 109/L was (2. 613 6 ± 1. 610 5)%. There were significant statistical difference(t= 3. 262, P<0.05). The possibility of viral replication became strong after the day 7 post-infection. There was a significant difference of viral replication between two groups in the day 7,10, 15 post-infection(t value was 3. 771, 2. 509 and 2. 260 respectively, P<0. 05). The possibility of viral replication was higher in CD4≤0.2 ×109/L group than that of CD4>0.2 × 109/L group. The logarithm of viral load was (5. 606 8 ± 0. 815 1 ) copies/mL in CD4≤0.2 × 109/L group and (4. 729 8 ± 0. 431 6) copies/mL in CD4> 0.2 × 109/L group. There was a significant difference between two groups(t = 3. 771 ; P<0.05). Conclusion Viral infection and replication are enhanced during progression of disease, even if viral coreceptor usage do not switch from CCR5 to CXCR4.
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Objective To investigate the effects of chemokine antagonist,Met-RANTES,on the acute rejection of islet allograft in the rat model.Methods According to the different treatments,rats were divided into 2 group: control group,allogeneic islet transplant untreated;experiment group,allogeneic islet transplant treated with Met-RANTES(200 ?g/day, i.p) for 7 days post-operation.The survival time of rat of islet transplant and blood sugar were recorded,and the pathological changes of islet allograft were observed.Scintillation counter was used to count the count per min(cpm) of monocytes.Flow cytometry was used to detect the ratio of CD4~+/CD8~+ phenotypes and CCR5 expression of peripheral blood lymphocytes.Results The mean survival time of islet allograft in experiment group was(23.0)?(10.5) days,obviously longer than in the control group((3.8)?(4.5) days,P
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BACKGROUND AND OBJECTIVES: CC chemokine receptor (CCR5) is characteristic of the Th 1 phenotype, the receptor of RANTES, MIP-1alphaand MIP-1beta. The receptor of CCR5 delta32 (a 32 bp deletion in the CCR5 gene, mutant type) results in the production of a non-functional receptor. Given the potential importance of CCR5 in allergic inflammation, we hypothesized that individuals carrying the CCR5 delta32 allele would show a reduced prevalence of allergic rhinitis. SUBJECTS AND METHOD: Blood samples for genetic analysis were obtained from 187 individuals with allergic rhinitis and from 278 healthy subjects without atopic diseases. Polymerase chain reaction-based assay for the CCR5 gene polymorphism was used for genotyping. RESULTS: We could not find the CCR5 delta32 homozygotes and heterozygotes at all in neither of the controls nor allergic rhinitis Korean patients. CONCLUSION: Since the CCR5 delta32 allele frequency did not deviate from that in the healthy control population, it is unlikely that this allele influences predisposition to allergic rhinitis in Koreans.
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Humans , Alleles , Asian People , Chemokine CCL4 , Chemokine CCL5 , Gene Frequency , Heterozygote , Homozygote , Inflammation , Phenotype , Prevalence , Receptors, CCR , Receptors, CCR5 , RhinitisABSTRACT
Since the first case of human immunodeficiency virus (HIV) infection in the Republic of Korea (ROK) was detected in 1985, 876 HIV-infected patients have been reported, as of December 1998. The male to female ratio was 6.8:1, and 87% of the patients were between 20 and 49 years of age. The major modes of transmission were sexual contacts, accounting for 86% of the cases (65% heterosexuals and 21% homosexuals). Transmission through blood and blood products accounted for 28 cases (3.2%), and vertical transmission for one case. No cases among intravenous drug abusers were reported. The seroprevalence among the blood donors was approximately one in 100,000. Subtypes A, B, C, D, E, and G of HIV-1 have been introduced into the ROK, and subtype B is the most predominant subtype. The frequency of the a deletion in the CCR5 gene, a coreceptor of HIV-1, was less than 1% among Koreans.