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1.
AIM: To explore the correlation between development of CD4+CD25+ regulatory T cells (CD4+CD25+ Tr) and thymus CD4-CD25+ cells. METHODS: The ratios of CD4+CD25+ regulatory T cells to CD4+ T cells in thymus, spleen, lymph node and peripheral blood of mice from birth to mature and also the ratios of CD4-CD25+ cells to CD4- T cells in thymus were measured by flow cytometry. Purified CD4+CD25+ T cells and CD4+CD25- T cells were labeled with CFDA-SE, and then stimulated with various kinds of stimulators. RESULTS: The percentages of CD4+CD25+ Tr in mouse spleen, lymph nodes and peripheral blood increased gradually, but not in thymus, from day one to week 10 of the age with rapid rising from day one to week 1. The percentages of CD4-CD25+ cells in mouse thymus were quite high on day one after birth, and decreased rapidly from day one to week 1. Both CD4+CD25+ Tr and CD4+CD25- T cells showed no proliferation in response to ConA, while CD4+CD25+ Tr showed a transient enlargement of cell size. Both CD4+CD25+ Tr and CD4+CD25- T cells underwent proliferation in response to PDB plus ionomycin. CD4+CD25- T cells, but not CD4+CD25+ Tr, showed a proliferative response to the stimulation of coated anti-CD3 plus soluble anti-CD28 antibody, however, CD4+CD25+ Tr showed significant proliferation and CD4+CD25- T cells showed a stronger response in addition of high dose of IL-2. CONCLUSION: The thymus CD4-CD25+ cells are probably the precursor of CD4+CD25+ Tr during cell development.  相似文献   

2.
AIM: To investigate the effect of glucocorticoid inhalation on the levels of CD4+CD25+ regulatory T cells in peripheral blood of asthmatic children. METHODS: Glucocorticoid inhalator was inhaled by 70 children with attack asthma. The levels of CD4+CD25+Tr in peripheral blood of asthmatic children were tested by flow cytometry (FCM). RESULTS: The CD4+CD25+Tr levels in peripheral blood of asthmatic children were (5.62%±1.29%) and (7.05%±1.61%) before and after of regulated glucocorticoid inhalation, respectively (P<0.01). The Tr levels were (7.56%±1.88%), (7.09%±1.23%) and (6.11%±1.96%) in the complete control group, part control group and poor control group, respectively (P<0.05). The Tr level in formal treatment group (7.05%±1.61%) was higher than that in irregular treatment group (5.91%±1.76%), P<0.01. CONCLUSION: The level of CD4+CD25+Tr is remarkable increased by regulated glucocorticoid inhalation, and the level of Tr can reflect the effects of glucocorticoid inhalation.  相似文献   

3.
AIM: To investigate the feasibility of inducing experimental autoimmune encephalomyelitis(EAE)model in C57BL/6 mice by TrxA-extracellular immunoglobulin domain of MOG(MOGIgd-TrxA)fusion protein produced by molecular cloning in our laboratory. Also to investigate the role of CD4+CD25+ T cells in the pathogenesis of EAE. METHODS: (1)The MOGIgd-TrxA fusion protein was induced and produced by molecular cloning and purified by metal chelate affinity chromatography and concentrated through ultrafiltration. The concentration of the protein was measured by Bradford method at last. (2)Animal experiment: C57BL/6 mice(12 mice in each group)were used. The mice in group MOG were immunized with MOGIgd -TrxA fusion protein. The mice in group GPSCH were received emulsion of spinal cord homogenate of guinea pigs(GPSCH), and mice in group TrxA or normal control group(group NC)were received the same volume emulsion of TrxA or saline/adjuvant, respectively. Clinical scores and histopathology were measured to value the models quality. (3)The percentages of CD4+CD25+ regulatory T cells in EAE mice were tested through flow cytometric analysis. RESULTS: (1)The purity of purified MOGIgd -TrxA fusion protein was about 98%, and its concentration was 2.3 g/L. (2)No significant difference between group MOG and group GPSCH in the clinical score was observed(P>0.05). Histologic sections of the brain and spinal cord taken from affected animals in both groups showed pathological change of different level throughout the central nervous system(CNS). (3)Percentages of CD4+CD25+ T cells in group MOG and group GPSCH were(4.71±1.61)%and(1.44±0.65)%,respectively, both of which were significantly lower than those in group NC(9.22±1.24)%and TrxA group(8.97±1.20)％(P<0.01). CONCLUSION: (1)The animal model of EAE in C57BL/6 mice induced by MOGIgd fusion protein produced through molecular cloning in our laboratory is stable and with high incidence. Thus, the author finds a good way to study the immune mechanisms of MS further and to search for the effective treatments as well. (2)The reduction of CD4+CD25+ T cells in EAE mice may have some relationship with the clinic.  相似文献   

4.
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5.
AIM:To confirm that CD4+CD25+ regulato ry T cells don't have an instinctive defection in IL-2 secretion,and to have an insight into the maturation state of CD4+CD25+ T cells in cord blood.METHODS:CD4+CD25+ and CD4+CD25- T cells were purified f rom cord blood of term infants (CB) and adult peripheral blood (PB) by autoMACS,and stimulated with PDB plus ionomycin.After 45 hours of culture,cells were d etected for expression of CD69 and CD25 by flow cytometry,and the supernatants were measured for 7 kinds of cytokines by Luminex.RESULTS:CD4+CD25+ T cells from both CB and PB proliferated comparably with CD4+CD25- T cells when stimulated with PDB plus ionomycin.A fter 45 hours of culture,however,the CD4+CD25+ T cells underwent a tendenc y of cell death.Expression of CD25 was further upregulated when CD25+ cells w ere activated.Under stimulation of PDB plus ionomycin,both CD4+CD25+ and C D4+CD25- T cells in PB secreted high levels of IFN-γ,IL-2 and TNF-α,with CD25+ cells secreted much higher level of IL-5,IL-4 and IL-10 than those in CD25- cells;CD4+CD25+ and CD4+CD25- T cells in CB also secreted high level of IL-2 and TNF-α but much lower level of IFN-γ than those in PB,and no secretion of IL-5,IL-4 and IL-10 was observed.CONCLUSION:CD4+CD25+ regulatory T cells don't have an i nstinctive defection in IL-2 secretion,otherwise there may be a different TCR signaling pattern in CD4+CD25+ T cells from traditional T cells.The CD4+C D25+ T cells in cord blood have not fully matured in function.  相似文献   

6.
AIM: The bacillus calmette-guerin (BCG) vaccine is the most widely used Th1-inducing vaccine. In recent years, some studies argued that mycobacterium vaccae can be used as adjuvant to induce regulatory T cells (Treg) and then suppress asthmatic airway inflammation. We previously have engineered recombined BCG that expressed Der p2 of house dust mites (Der p2 rBCG) on the cell wall. The aim of this study is to investigate the immune regulatory mechanisms of Der p2 rBCG. METHODS: Mice were vaccined with PS, BCG or rBCG. The relative proportion and the absolute numbers of related Tregs in spleen cells were analyzed. The suppressive activity of Der p2 rBCG-induced CD4+CD25+ T cells was detected both in vitro and in vivo. RESULTS: (1) Der p2 rBCG induced a CD4+CD25+Foxp3+ T cell subtype. (2) Der p2 rBCG-induced CD4+CD25+ T cells suppressed the proliferation of Th2 effector cells in vitro in an antigen-specific way. (3) Der p2 rBCG-induced CD4+CD25+ T cells mediated Der p2 specific suppression of airway allergy in vivo. CONCLUSION: Der p2 rBCG induces a CD4+CD25+Foxp3+ T cell subtype, which suppresses inflammation in allergic airway in a mouse model.  相似文献   

7.
AIM: To investigate the effect and mechanism of Foxp3-transduced CD4+CD25-T cells on the cytotoxicity of NK cells. METHODS: Retroviral Foxp3 gene transfection was applied to nave CD4+CD25-T cells. Fresh transduced CD4+Foxp3+ T cells were co-cultured with NK cells. ［51Cr］ labeled YAC-1 cells were used to detect NK cells cytotoxicity. The anti-TGF-β antibody was added into the co-culture system to detect the TGF-β blocking effect. Also the transwell co-culture system was used to investigate the regulatory effect of Treg cells on NK cells. RESULTS: One week after transduction, 38.0% of Foxp3-transduced T cells showed GFP expression by flow cytometry. Foxp3-transduced CD4+CD25-T cells suppressed function of NK cells. The inhibition rates of Foxp3 transduced CD4+CD25- T cells were 42.9% at 24 h and 22.7% at 48 h. When anti-TGF-β antibody was added to the co-culture system, the inhibition rate of CD4+Foxp3+ T cells was 3.2% and 2.1%, respectively. CONCLUSION: CD4+Foxp3+ T cells significantly inhibit the cytolytic function of NK cells. TGF-β plays different roles on this action in different inhibition systems. The inhibitory effect of Treg cells on NK cells is cell-to-cell contact dependent and associates with TGF-β expression.  相似文献   

8.
AIM：To investigate new methods for the expansion of human CD8+ memory T cells in vitro and provide novel means of anti-viral and anti-tumor adoptive immunotherapy. METHODS：Six kinds of stimuli, anti-CD3 antibody, anti-CD28 antibody, CD70, IL-2, IL-7 and IL-15, for the expansion of human CD8+ memory T cells in vitro were selected and arranged for their combinations, resulting in 63 kinds of stimulating combinations. Normal human CD8+ T cells were isolated and exposed to these stimuli. After 14 days of cell culture, the number and purity of CD8+ T cells, and the percentages of CD8+ central memory T cells (TCM) and CD8+ effector memory T cells (TEM) were detected. The expansion folds of CD8+ T cells, CD8+ TCM and CD8+ TEM were calculated and the relatively better stimulating combination was determined. RESULTS：The combination of anti-CD3 antibodies, IL-2 and IL-7 was a better method for the expansion of human CD8+ T cells, CD8+ TCM and CD8+ TEM in vitro, and the expansion folds were 13.19, 13.28 and 15.27, respectively. CONCLUSION：The combination of anti-CD3 antibodies, IL-2 and IL-7 is the relatively better method for the expansion of human CD8+ memory T cells in vitro.  相似文献   

9.
AIM: To analyze the possibility that volume-activated chloride channel (VACC) exists in tumor stem cells. METHODS: CD133+ cells were purified by magnetic cell separation (MACS) system from non-small-cell lung cancer cell line A549. The purity of the cells was detected by flow cytometry. VACC current was recorded with whole-cell patch-clamp. RESULTS: The purity of CD133+ cells isolated from A549 by MACS was 92.14%. VACC current was recorded in the 22/40(55%) CD133+ cells of A549 by whole-cell patch-clamp. CONCLUSION: VACC exists in CD133+ lung cancer stem cells.  相似文献   

10.
AIM:To observe the apoptosis and the expression of forkhead box protein 3(Foxp3) induced by magnesium in CD4+CD25+ regulatory T cells isolated from healthy and asthmatic human peripheral blood. METHODS:Peripheral blood from healthy volunteers and asthma patients was collected. CD4+CD25+ T cells were separated by Percoll centrifugation and magnetic separation. The cells were cultured for 72 h and treated with magnesium(10 mmol/L) or control solution. The apoptotic rate and the expression of Foxp3 in the cells were analyzed by flow cytometry. RESULTS:The purity of CD4+CD25+T cells was 77.4%~92.3% in health group, and was 75.2％~93.8％in asthma group. The proportion of CD4+CD25+ T cells in CD4+T cells was 4.12%~7.98% in healthy adults, and 4.51%~8.68% in asthma patients. No significant difference between the 2 groups was observed. Magnesium at concentration of 10 mmol/L up-regulated the apoptotic rate of CD4+CD25+ T cells(P<0.05) and did not affect the Foxp3 expression in the cells in both health and asthma groups. CONCLUSION:Magnesium plays therapeutic effects on asthma by inducing the apoptosis of peripheral CD4+CD25+ regulatory T cells.  相似文献   

11.
AIM: To analyze the effects of oxymatrine (OMT) on the quantity of murine regulatory T cells (Tr cells) in the peripheral blood and mouse lymphocyte proliferation stimulated by Con A, and to probe into the immunological mechanism that OMT treats allergic contact dermatitis (ACD).METHODS: An ACD mouse model stimulated by dinitrofluorobenzene (DNFB) was established. Different dosages of OMT, PBS and hydrocortisone (HCT) were intraperitoneally injected (IP) into the mice. Blood samples were collected at〖JP+2〗 1 d, 7 d, 14 d, 21 d and 28 d, then the T cells were isolated and marked with anti-CD3, anti-CD4, anti-CD25 three-colored immune fluorescence antibody to detect the quantity of CD4+CD25+ T cells with flow cytometry. The fluorescence intensity changes of lymphocytes which were isolated from mouses lymph node and co-stimulated by polyclonal stimulator Con A and OMT were examined by carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) staining and flow cytometry. RESULTS: OMT at concentrations of 500, 125 and 31 mg/L had the ability to restrain the proliferation of lymphocytes from lymph node in a dose dependent manner. However, OMT at concentrations of 16, 8, 4 and 2 mg/L promoted the proliferation of T lymphocytes from lymph node, but was not obviously dependent on its concentration. Intraperitoneal injection of OMT increased the numbers of CD4+CD25+T cell in peripheral blood obviously (P<0.01). CONCLUSION: The effects of OMT on the proliferation of T lymphocytes from mouses lymph node cells are observed, OMT also increases the CD4+CD25+T cells in the peripheral blood, implying that OMT is a kind of immunoregulator with dual effects.  相似文献   

12.
AIM: To investigate the function of caspase-3 and mitogen-activated protein kinases (MAPKs) in allogeneic CD8+T cell-induced apoptosis of vascular endothelial cells. METHODS: Allogeneic CD8+T cells were isolated from PBMC by positive selection using magnetic beads coated with anti-CD8 antibody. After cocultured with allogeneic CD8+T cells, apoptosis of human umbilical vein endothelial cells (HUVECs) and human dermal microvascular endothelial cells (HDMECs) were detected by AnnexinV-FITC labeling. Western blotting was used to examine the change of MAPK and caspase-3 expression in the vascular endothelial cells. The influence of SB203580 (inhibitor of p38MAPK), SP600125 (inhibitor of JNK), PD98059 (inhibitor of ERK), Z-DEVD-FMK (a caspase-3-specific peptide inhibitor) on apoptosis was also examined. RESULTS: At 24 h and 48 h time-point, the apoptosis rates of HUVECs were 41.7%±10.1% and 29.4%±8.3%, respectively (P<0.01, vs untreated HUVECs); the apoptosis rates of HDMECs were 28.9%±7.2% and 15.2%±4.8%, respectively (P<0.01, vs untreated HDMECs). These effects were largely prevented by Z-DEVD-FMK and SB203580 (P<0.05). Allogeneic CD8+T cells enhanced cleavage of caspase-3 and led to p38MAPK phospholation. CONCLUSION: Caspase-3 and p38MAPK mediate allogeneic CD8+T cells-induced apoptosis of vascular endothelial cells.  相似文献   

13.
AIM: To investigate the effect of lipoxin A4 on the proliferation and IL-2 production in Jurkat T cells. METHODS: Jurkat T cells were activated in vitro with anti-CD3 (2 mg/L) and anti-CD28 (2 mg/L) antibodies in the absence or presence of lipoxin A4 (0.1 nmol/L-100 nmol/L) for 24 h, then ［3H］-TdR was added into the medium and radioactivities were measured by scintillation counting. The concentrations of IL-2 in the supernants were determined by ELISA. Cells were harvested and the expression of CD25 was assessed by FCM. For analysing the cell cycle, the cells were stained with PI and DNA contents were detected by FCM. RESULTS: Lipoxin A4 suppressed the proliferation of anti-CD3 and anti-CD28 antibodies activated Jurkat cells in a dose-dependent manner, which was associated with reduced proportion of S phase cells. Furthemore, lipoxin A4 significantly inhibited the production of IL-2 but had no obvious effect on CD25 expression. CONCLUSION: Lipoxin A4 can suppress proliferation of activate Jurkat cells and IL-2 production, through which lipoxin A4 might negatively regulate immune response.  相似文献   

14.
AIM：To investigate the effect of vitamin C (VC), a small-molecule compound, on the expansion of CD4+ effector memory T cells (TEM) in vitro and to improve the effect of adoptive immunotherapy. METHODS：Normal human peripheral blood CD4+ T-lymphocytes were isolated and randomly divided into 2 groups: control group and experiment group. VC was added into experimental group. Thereafter, the expansion of CD4+ TEM in the 2 groups was detected by cell counter and flow cytometry. RESULTS：VC did not significantly affect the total number of CD4+ T cells, while it raised the ratio of TEM in CD4+ T cells at an optimal concentration of 100 mg/L. After 10 days of the expansion of CD4+ T cells, the number of CD4+ TEM in experiment group was (3.56±0.35)×106, significantly larger than that in control group ［(1.22±0.15)×106, P<0.01］. CONCLUSION: VC can effectively promote the expansion of CD4+ TEM in vitro, which provides a simple, safe and effective expansion method for adoptive immunotherapy.  相似文献   

15.
AIM: To investigate the fraction of CD4+CD28-T cells and its correlation with lymphocytic apoptosis in peripheral blood of rheumatoid arthritis (RA) patients.METHODS: The RA patients and age-matched health controls were selected in the study. The lymphocytes were isolated from peripheral blood. CD4+ T cells without CD28 expression (CD4+ CD28-) were analyzed by flow cytometry. The incidence of apoptosis in the cells cultured with or without PHA for 24 h was determined by flow cytometry with Annexin V-FITC/PI di-staining. The correlation between the fraction of CD4+CD28-T cells and lymphocytic apoptosis was also observed.RESULTS: The fraction of CD4+CD28-T cells was significantly higher in RA group than that in healthy control group (7.79%±3.52% vs 1.89%±1.78%, P<0.05). The apoptotic level in PHA cultured lymphocytes was significantly lower in RA group than that in healthy controls (11.38%±5.73% vs 19.46%±6.32％, P<0.05). Spearman analysis showed that there was a negative correlation between the fraction of CD4+CD28-T cells and apoptotic level of activated lymphocytes (r=-0.433,P<0.01).CONCLUSION: The increased CD4+CD28-T cells contribute to prolong the lifespan of activated lymphocytes in peripheral blood of RA patients, and the persistence of activated lymphocytes may contribute to the pathogenesis of RA.  相似文献   

16.
AIM: To investigate the pattern of Th1/Th2 balance in systemic lupus erythematosus(SLE) patients and the relationship between CD28/CTLA-4(cytotoxic T-lymphocyte antigen-4) molecule expression and Th1/Th2 balance.METHODS: Eighteen SLE patients met the ARA 1997 updated SLE criteria were selected in the study. According to Bombardier's SLEDAI criteria, all patients were classified into two groups: active group(12 cases) and static group(6 cases). Fourteen normal individuals, matched for age and sex of the patients, served as controls. The peripheral blood mononuclear cells(PBMCs) were isolated by density gradient centrifugation and cultured in RPMI-1640 culture medium. After treated with PMA(5 μg/L) and ionomycin(500 μg/L) for 72 h, the PBMCs were collected, the contents of IFN-γ and IL-10 in the supernatant of cultured PBMCs were detected using enzyme linked immunosorbent assay(ELISA). The expression of CD28 and CTLA-4 molecules on T cells were detected by flow cytometric technique with double staining by FITC or PE labeled monoclonal antibodies. RESULTS: The level of IL-10 was higher in the PBMCs of active and static SLE patients(351.29 ng/L±153.31 ng/L and 319.37 ng/L±153.39 ng/L) than that in controls(254.48 ng/L±120.69 ng/L), but the difference did not reach statistical significance(P>0.05). The level of IFN-γ was significantly lower in the PBMCs of active SLE patients(25.76 ng/L±16.09 ng/L) than that in controls(50.71 ng/L±27.92 ng/L, P＜0.05). The ratio of IL-10/IFN-γ was significantly higher in active SLE patients(18.74±13.77) than that in controls(6.66±4.95, P<0.05). Either before or after culture, the expression of CD28 molecule on CD3+and CD8+ T cells from all SLE patients was not remarkably different from that in the cells of controls. Before culture, the expression of CTLA-4 molecule on CD3+T cells of active SLE patients(0.79%+0.37%) was significantly lower than that in the cells of controls(1.31%+0.61%, P<0.05). After culture, the expression of CTLA-4 molecule on CD3+ T cells of SLE patients was still lower than that in the cells of normal controls without statistical significance(P＞0.05).The expression level of CD28 molecule on CD3+ or CD8+ T cells in active SLE patients and controls was not correlated with the levels of IFN-γ and IL-10 in the supernatants(P＞0.05). The level of CTLA-4 molecule expression on CD3+ T cells of active SLE patients was positively correlated with IFN-γ level(r=0.681, P<0.05), while was negatively correlated with IL-10 levels(r=-0.624,P＜0.05) and the ratio of IL-10/IFN-γ(r=-0.738, P<0.01). The level of CTLA-4 molecule expression on CD3+ or CD8+ T cells of controls showed no correlation with IFN-γ levels, while showed negative correlations with IL-10 level(r=-0.587, P<0.05; r=-0.563, P<0.05, respectively).CONCLUSION: There is a bias in the differentiation of Th0 cells towards Th2 in SLE patients. CTLA-4 probably plays an important role in this mechanism through suppressing the signal transmitted by CD28.  相似文献   

17.
AIM: To investigate how human adipose-derived stem cells (hASCs) regulates the differentiation of Th17 cells in multiple sclerosis. METHODS: hASCs were isolated from the adipose tissues. Magnetic-activated cell sorting (MACS) kit was used to isolate CD4+ T cells from peripheral blood mononuclear cells (PBMCs) which were isolated by density gradient centrifugation. The percentage of CD4+ T cells was detected by flow cytometry. The activated CD4+ T cells were co-cultured with hASCs for about 4 d at different ratios of hASCs to CD4+ T cells (1:4 and 1:10) in a Th17 polarised condition. Another group adding anti-leukemia inhibitory factor (LIF) antibody was set up. Th17 cell proportion of the CD4+ T cells was determined by flow cytometry. The level of LIF in the supernatant of co-cultured system was measured by ELISA. The mRNA expression of retinoid-related orphan receptor γt (RORγt), interleukin-6 receptor (IL-6R), interleukin-23 receptor (IL-23R), LIF and leukemia inhibitory factor receptor (LIFR) was detected by real-time PCR. RESULTS: The result of flow cytometry suggested there were mainly hASCs, and the percentage of CD4+ T cells in the PBMCs were above 90% after MACS. The Th17 cell proportion decreased in 1:4 and 1:10 co-cultured groups in a dose-dependent manner. The mRNA expression of IL-6R, IL-23R and RORγt was downregulated and the expression of LIFR and LIF was up-regulated. When the anti-LIF was added into the co-cultured system, the ratio of Th17 cells increased and reached to the control level. The protein level of LIF obviously increased after co-cultured. After anti-LIF added, the mRNA expression of RORγt and IL-6R was up-regulated. CONCLUSION: hASCs inhibits the differentiation of Th17 cells from multiple sclerosis patients through the competitive inhibition of LIF/IL-6 by secreting LIF.  相似文献   

18.
AIM: To investigate the role of Th17 cells in the patients with cervical cancer.METHODS: We measured the peripheral levels of Th17 cells and CD3+CD8-IL-21+ T cells in 37 cervical cancer (CC) patients, 25 cervical intraepithelial neoplasia (CIN) patients and 18 healthy controls by flow cytometry. The percentages of Th17 cells and CD3+CD8-IL-21+ T cells in total CD4+ cells were calculated.RESULTS: Compared with controls, the patients with CC or CIN had higher proportions of Th17 cells (all P<0.01) and CD3+CD8-IL-21+ T cells (all P<0.05). Notably, in CC patients, the increased percentages of Th17 cells and CD3+CD8-IL-21+ T cells were independently associated with the clinical stage(all P<0.05), lymph node metastasis (P<0.01,P<0.05) and vasoinvasion (all P<0.01), while the elevated percentage of CD3+CD8-IL-21+ T cells was also associated with the tumor size(P<0.01). Furthermore, the percentage of Th17 cells was positively correlated with that of CD3+CD8-IL-21+ T cells in healthy controls and CC patients, but not in CIN patients.CONCLUSION: Our results indicates a possible role of Th17 cells in CC patients correlated with CD3+CD8-IL-21+T cells, and the elevated percentage of circulating Th17 cells may be involved in the development and progression of cervical cancer.  相似文献   

19.
AIM：To observe the effect of acupuncture on CD4+ CD25+ Foxp3(forkhead box P3)+ regulatory T-cells(Treg cells) in rats with embryo implantation failure. METHODS：One hundred and forty-four pregnant rats were randomly divided into control group(N), mifepristone treatment group(M), mifepristone+acupuncture treatment group(A) and mifepristone+progestin treatment group(W). The rats in groups M, A and W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. The Housanli(ST36) and Sanyinjiao(SP6) points were selected for acupuncture. From day 1 to the time of death, the rats in group A were fasten up and then the acupuncture was performed. Accordingly, the rats in group N and group M were only fixed, and the rats in group W were given progestin. Implanted embryos in each group were counted. The proportions of CD4+ CD25+ Foxp3+ Treg cells in peripheral blood and CD4+ Foxp3+ Treg cells in the endometrium were detected by flow cytometry. The mRNA and protein levels of Foxp3 were determined by real-time PCR and Western blotting, respectively. RESULTS：Compared with group N, the number of implanted embryos, the percentages of CD4+ CD25+ Foxp3+ Treg cells in peripheral blood and CD4+ Foxp3+ Treg cells in the endometrium, and the expression of Foxp3 protein and mRNA in the endometrium were significantly decreased in group M(P<005). Compared with group M, the above indexes in group A and group W were significantly increased. CONCLUSION: The effect of acupuncture in rats with embryo implantation failure may be closely correlated with the modulation of CD4+ CD25+ Foxp3+ Treg cells.  相似文献   

20.
AIM: To isolate, purify and differentiate endothelial progenitor cells from cord blood in vitro and to study their biological characteristics. METHODS: CD133+ cells were selected from fresh cord blood mononuclear cells (MNC) by magnetic activated cell-sorting system (MACS). EPC was studied by flow cytometry, immunocytochemistry and immunofluorescence staining. Isolated cells were cultured in IMDM medium supplemented with or without VEGF, bFGF, SCF. RESULTS: The percentage of CD133+ cells of cord blood MNC was (1.41±1.14)%, and purity was 75%-85% (FACS method). CD133+ cells were grown on fibronectin-coated chamber slides in the presence of VEGF, bFGF, SCF. Within 1-2 hours of culture cells became adherent. On day 7-10, the adherent cells displayed a typical “cobblestone” morphology. After 14 days of culture, the adherent cells revealed a heterogeneous cell population, comprising small-sized round cells, spindle-like cells and formed tube-like structure. Weibel-Palade bodies were shown on the transmission electron microscopy photomicrographs. Compared with the original, cell markers CD133 and CD34 decreased significantly (77.0%±3.3% to 1.6%±2.2% and 93.1%±4.7% to 37.4%±4.9%, P<0.05), while Flk-1 increased significantly (from 22.3%±3.3% to 94.3%±4.1%, P<0.05) after 14 days of culture with VEGF, bFGF, SCF. The vWF was strongly expressed (77.9%±3.3%) on the 14th day later. CONCLUSION: Vascular endothelial progenitor cells were isolated from cord blood with specific expression of CD133/CD34/Flk-1. With the stimulation of the growth factors, seven-ten days after culture EPCs could be turned to endothelial cells.  相似文献   

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