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1.
AIM: To determine whether Sca-1+ cells from fetal liver can differentiate into neural cells. METHODS: The sex of 14.5-day-old murine fetuses was determined by PCR analysis of sry gene, and Sca-1+ cells from male fetal liver were isolated with a magnetic cell sorting kit, 2×103 of which were then transplanted into lethally irradiated female mice. The donor cells and their characteristics in recipient brains were identified and detected by FISH and immunohistochemistry double-staining analysis at 60, 120, 180 days after transplantation. RESULTS: There existed many male cells in brains of female recipients, some of them express neuron-specific nuclear protein (NeuN), and some of them express the astrocyte-specific marker, i.e. glial fibrillary acidic protein (GFAP). CONCLUSION: Sca-1+ cells from fetal liver, which contain hematopoietic stem cells, can differentiate into neuronal cells and astrocytes in the brains of adult mice.  相似文献   

2.
LIU Ge-xiu  ZHANG Yuan 《园艺学报》2003,19(9):1178-1181
AIM: To study whether Sca-1+ cells from fetal liver can be induced to differentiate into neuronal cells in vitro. METHODS:Sca-1+cells from 14 5-days-old murine fetal liver were isolated with a magnetic cell sorting kit, and were cultured in Dulbecco s modif ied Eagle s medium(DMEM)/F12 supplemented with 10%fetal bovine serum(FBS), and passaged at a rat io of 1 3 when cells reached more than 80%confluence.The 5 passage cells were induced by 10-3mol/Lβ-mercaptoethanol(β-ME)and 5×10-7 mol/L all-trans-retinoic acid(RA)for 24 hours, and then incubated in serum-free medium for 5 hours to 5 days.The characteristics of treated cel s were assayed by immunocytochemistry staining analysis at 5 hours, or 5 days.RESULTS: Cells treated with β-ME and RA exhibited neuronal phenotype and expressed neuron-specific protein such as neuron-specific nuclear protein (NeuN), neuronfilament-M, and neuron-specific tubulin-1 (TuJ-1) but not tau, MAP-2, or the astrocyte-specific marker glial fibrillary acidic protein (GFAP).CONCLUSION: Sca-1+ cells from fetal liver, of which most are regarded as hematopoietic stem cells, could differentiate into early immature neuronal cells in vitro. These findings suggest that Sca-1+ cells from fetal liver may be an alternative source in cell therapy and gene therapy of neural dysfunction.  相似文献   

3.
AIM: To investigate the differentiation of murine embryonic stem cells (ESCs) into hematopoietic stem cells (HSCs) by the supportive effects of human aorta-gonad-mesonephros (AGM) region and fetal liver (FL) stromal cells.METHODS: E14 ESCs were induced into embryoid body (EB) first. Then the cells from EB were further co-cultured with human AGM region and FL stromal cells in non-contact system. On day 6, the cells derived from EB were collected for Sca-1+c-Kit+ cell analysis by flow cytometry, colony forming unit (CFU) assay and teratoma formation checking. BALB/c female mice conditioned with lethal dose of γ-ray irradiation were transplanted with EB cells from different culture systems. The survival rates, engraftment of donor cells, reconstitution of hematopoietic were monitored.RESULTS: Sca-1+c-Kit+ cells in EB cells co-cultured with human AGM region and FL stromal cells had the value of (21.96±2.54)%, and the total CFU was as (520±52)/105 cells, which were statistically greater than those in EB cells only cultured with human AGM region stromal cells (P＜0.05). No teratoma was found in NOD-SCID mice after subcutaneous injection of EB cells co-cultured with human AGM region and FL stromal cells. In BALB/c female mice transplanted of EB cells co-cultured with human AGM region and FL stromal cells, the survival rate was 77.8%, and the peripheral blood cell count was obviously improved on day 14. PCR results showed the recipients all had sry gene copies from donor in bone marrow. The recipient mice transplanted with EB cells only cultured with human AGM region stromal cells all died within 15 days.CONCLUSION: Stromal cells from human AGM region and FL enhance the directed differentiation of ESCs into HSCs which can reconstruct hematopoiesis in vivo.  相似文献   

4.
AIM: To study the effect of acute renal failure (ARF) on the differentiated frequency of Sca-1+ cells from murine fetal liver in irradiated mice. METHODS: The Sca-1+ cells from murine fetal liver were isolated with magnetic cell sorting (MACS) technique, the sex of which was identified by PCR. The 2×104 Sca-1+ cells were transplanted into a lethally irradiated (［60Co］, 8 Gy) inbred female mouse. After 8 weeks, these recipient mice were divided to A, B, and C groups at random (A group: irradiated; B group: ARF; C group: ARF and Sca-1+). The mice in B and C groups were induced to ARF with 50% (V/V) glycerin (11.6 mL/kg). 72 hours later, the mice in C group were injected with the fresh prepared Sca-1+ cells again. 8 weeks later, mice were sacrificed, and their kidneys were taken out, fixed and slices were prepared. Fluorescence in situ hybridization (FISH) of renal slices was performed and the pictures of them were taken and analyzed. RESULTS: The cells containing Y chromosome were found in renal slices from the mice in A, B and C groups, which located in epithelial cells of renal tubules, interstitium, glomeruli, and glomerular margin and increased gradually. The double and encircle zone of Y chromosome cells were found in the slices from the mice in B and C groups separately, which was consist of new renal tubules. The differentiation frequency of Sca-1+ cells in kidney in A, B and C groups were (1.65±0.18)%， (8.58±1.34)% and (18.13±1.91)%， respectively, which showed significant difference between former group and later group (P<0.01). CONCLUSION: The differentiation of Sca-1+ cells from murine fetal liver into renal cells and tissue is promoted by physiological microenvironment of acute damage and regeneration.  相似文献   

5.
AIM: To study the relationship between tissue irradiation injury and past-transplantation distributing diversity of Sca-1 positive cells from murine fetal liver and their offspring cells in multi-organs after syngeneic sex mismatch hematopoietic remodeling.METHODS: The Sca-1 positive cells from the livers of C57BL/ 6j male mouse fetus aged 14.5 day were identified and separated by quick PCR and magnetic cell sorting (MACS) techniques. In order to reconstruct hematopoiesis of the adult female mice, which were lethally irradiated with 8G of ［Co60］, the 2×104 of Sca-1+ cells were transplanted through caudal vein into each of them. After 6 months, these recipient mice were sacrificed, and their kidneys, livers, lungs, stomachs, and small intestines were taken out, fixed and slices were made. Fluorescence in situ hybridization was performed and observed by fluorescent microscope. Images were captured and analyzed through appropriative cool CCD and software. RESULTS: After 2×104 of Sca-1+ cells were transfused, the hematopoietic function in the lethally irradiative female mice was completely restored. The cells containing Y chromosome were observed 6 months latter in multi-organs, including kidney, liver, lung, stomach, and small intestine. The frequency of the cells containing Y chromosome respectively was kidney (1.65±0.18)%, liver (0.90±0.10)%, lung (1.90±0.60)%, stomach (6.10±0.50)%, and small intestine (7.61±2.30)%, presented the trend of small intestine>stomach>lung>kidneys>liver. Combined informational analysis showed that the presenting frequency of the cells containing Y chromosome was consistent with the irradiative sensitivity of the organ. CONCLUSION: These findings suggest that the capability of differentiation of Sca-1 positive cells from murine fetal liver was potentially connected to the extent of damage in these organs when transferred in vivo.  相似文献   

6.
AIM:To study the effect of adoptive transfer of CD4+ T cells with microRNA-7 (miR-7) knockdown (KD) on mouse acute liver injury model and to investigate its significance. METHODS:CD4+ CD62L+ T cells were purified from the spleen of normal wild-type (WT) mice and miR-7KD mice by magnetic bead sorting, and were stained with CFSE. These 2×106 CFSE-labeling cells were injected into normal mice via tail vein, and then the mouse acute liver injury model was induced by intraperitoneal injection of 30 mg/kg concanavalin A. After 72 h, the appearance, weight and weight index of the liver were investigated. The pathological change of the liver tissues was observed by HE staining. Real-time PCR was used to examine the mRNA expression of Bax and P53. The expression levels of CD62L, interleukin-4 (IL-4) and interferon-γ (IFN-γ) in the CD4+ T cells were analyzed by flow cytometry. RESULTS:We found that the liver tissue became lighter, and the weight (P<0.01) and weight index (P<0.05) were changed significantly in miR-7KD mice compared with control group. Moreover, HE staining showed that the liver cell damage was increased in the liver of miR-7KD mice. Meanwhile, the expression levels of Bax and P53 were significantly increased in miR-7KD group (P<0.05). The percentage of CD62L in CD4+ T cells was significantly decreased (P<0.01) in miR-7KD mice, with high expression of IFN-γ (P<0.05) and low expression of IL-4 (P<0.01) in CD4+T cells. CONCLUSION:These findings suggest that miR-7 knockdown significantly promotes the pathology of CD4+ T cell-mediated acute liver injury, which provides a preliminary experimental basis for further exploration on the mechanism of acute liver injury occurrence.  相似文献   

7.
AIM and METHODS: To study the efficacy of the decoction of invigorating kidney and activating blood circulation(DIKABC) in BALB/C mice with immune mediated aplastic anemia. 50 female BALB/C mice at 8~12 weeks of age were divided into 5 groups:(1) control,(2) model,(3) model+DIKABC(H), (4) model+DIKABC(L), and (5) model+ciclosporin A(CsA). The model of immune mediated aplastic anemia in BALB/C mice, which received sublethal whole body irradiation, lymph node and thymus cells of DBA/2 mice, was given different liquid from groups and administered appropriately for 9 days, blood cells and bone marrow nucleated cells(BMNC) and CD45+ cells were determined by flow cytomytry(FCM).RESULTS: RBC,WBC,Platelet,BMNC in DIKABC groups were higher than those in model group (P＜0.01/0.05), but those in CsA group were lower than those in DIKABC groups (P＜0.05).Compared with model group, CD45+ cells in DIKABC groups increased (P＜0.05),but were lower than those in control groups (P＜0.05). CONCLUSION: The decoction of invigorating kidney and activating blood circulation could increase the percentages of CD45+ cells in bone marrow and prevent immune mediated aplastic anemia in BALB/C mice.  相似文献   

8.
AIM: To investigate whether aorta-derived CD105+ cells show characteristics of mesenchymal stem cells, and if dexamethason enhances this kind of CD105+ cells to differentiate into adipocytes. METHODS: The distribution of CD105 in aorta was assessed by imunohistochemistry. The aorta wall cells were isolated and immunophenotypes were identified by FACS. CD105+ cells were sorted using MACS CD105 micromagnetic beads. The differentiation of CD105+ cells into adipocytes and osteoblasts was induced under different conditions and indicated by staining of Oil red O, detecting of alkaline phosphatase activity, calcium accumulation stained with silver nitrate and transmission electron microscope analysis, respectively. RESULTS: The endothelial cells, a part of medial smooth muscle cells and adventital fibrblasts were CD105 positive. The isolated aortic arch cells were positive for CD105, CD106, CD44, CD29, and negative for CD45, CD11a, CD11b and HLADR. The CD105+ cells differentiated into adipocytes contained Oil-Red-O-positive lipid droplets, the osteocytes with calcium deposition and alkaline phosphatase activity. Ultrastructurally, it was observed that some needle-shaped crystal calcium deposition similar to bone spicules was inside the cytoplasm of induced osteocytes. When the dexamethason was absent in the adipogenic medium, there were no adipocytes with lipid droplets. CONCLUSION: The results demonstrated that CD105+ cells showed characters of MSCs reside in aortic wall, and was able to differentiate into adipocytes and osteocytes in vitro. Dexamethason enhanced aorta-derived CD105+ with characters of MSCs to differentiate into adipocytes. These suggested that MSCs might be related with atherosclerosis.  相似文献   

9.
AIM: To study the mechanism of butylated hydroxyanisole-induced neural differentiation of fetal liver Sca-1+ cells in vitro. METHODS: Sca-1+ cells from 14.5-day-old mouse fetal liver were isolated with a magnetic cell sorting kit. Cultured cells pretreated with or without extracellular signal-regulated kinase (MEK1) inhibitor, PD98059, were induced by 200 μmol/L butylated hydroxyanisole (BHA) for 24 hours, and then incubated in serum-free medium. Expression of genes in treated or untreated cells were assayed by Western blotting and RT-PCR. RESULTS: There was low level of neuronfilament-L (NF-L) and brain factor-1 (BF-1) in fetal liver Sca-1+ cells, but no neuronfilament-H (NF-H) and tyrosine hydroxylase (TH) was observed. BHA significantly promoted the expression of neuron-specific NF-L, NF-H, BF-1, and TH in fetal liver Sca-1+ cells. NF-L, NF-H, BF-1 and TH increased by 6.32 fold, 2.73 fold, 3.37 fold and 2.68 fold, respectively (all P<0.01 vs untreated cells). However, the expression of BHA-induced genes were inhibited by ERK kinase inhibitor PD98059 (25 μmol/L). CONCLUSION: BHA induces neural differentiation of fetal liver cells through ERK.  相似文献   

10.
AIM:To induce lymphoid stem cells and/or T-cell precursors to differentiate into functional mature T lymphocyte, and to increase the surface marker of T lymphocytes such as CD3+, while embryonic stem(ES) cells differentiated into hematopoietic stem/progenitor cells(HSPCs) in vitro. When they were injected into lethally irradiated mice, these differentiated cells had the advantage in immune reconstitution. METHODS:Embryonic stem cells formed embryoid bodies(EBs) in the medium containing methycellulose, hematopoietic growth factors(HGFs) was added to the culture system on the 6th day, thymopeptide was added at the same time. Flow cytometry were performed to detect the surface marker CD34+ and CD3+ of the differentiated cells. Finally the differentiated cells were injected into lethally irradiated mice, 60 days later, the incidence rate of graft versus host disease(GVHD) was taken as the mark of cell mediated immunity, PCR was performed to detect the sex determining region of the Y-chromosome(Sry) in bone marrow cells and spleen cells of the survival host female mice. RESULTS:The percentage of CD3+ T lymphocytes was 10.52% and the incidence rate of GVHD was 0% on the 13th day, but they respectively rose up to 22.93% and 100% if thymopeptide was added in the procedure of inducing ES cells to differentiate into HSPC in vitro. CONCLUSION:The quantity of CD3+ T lymphocytes increased in medium containing thymopeptide when ES cells differentiated into CD34+ HSPC.  相似文献   

11.
AIM:To investigate the effects of glucocorticoid on the regulation of microRNA-155 (miRNA-155) expression in the CD4+ T cells of asthmatic mice. METHODS:The ovalbumin (OVA)-induced asthma mouse model was established and the mice were treated with glucocorticoid. The effects of glucocorticoid on the pulmpnary histopathological changes, the expression of miRNA-155 in the lung tissues and CD4+T cells, and the levels of cytokines in the bronchoal-veolar lavage fluid (BALF) were evaluated. RESULTS:The results of RT-qPCR showed that the expressions of miRNA-155 in the lung tissues and CD4+T cells from the spleen of asthmatic mice were significantly increased, and the level of miRNA-155 in the CD4+T cells was significantly increased with the increase in the allergen exposure time (P<0.01). HE and PAS staining showed that OVA significantly increased inflammatory cell infiltration as compared with control group, and the peribronchial and perivascular inflammation and mucus secretion of proliferative goblet cells were significantly reduced after glucocorticoid treatment. Glucocorticoid treatment inhibited the increase in the proportion of CD4+ CD8- cells in the spleen and decreased the accumulation of CD4+ T cells in the lung tissues of asthmatic mice (P<0.01). After glucocorticoid treatment, the levels of interleukin-4 (IL-4), IL-5 and IL-13 in BALF were decreased, while the level of interferon-γ was increased significantly (P<0.01). CONCLUSION:Glucocorticoid reduces the accumulation of CD4+ T cells and inhibits the expression of miRNA-155 in the lung tissues and spleen CD4+ T cells of asthmatic mice.  相似文献   

12.
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   

13.
AIM: To investigate the biological characterics of human second-trimester fetal cord blood mesenchymal stem cells (MSC) and its application prospects in utero gene transfer/therapy (IUGT). METHODS: Nuclear cells separated from cord blood were cultured in DMEM medium. Surface antigens of the MSC were analyzed by the FACScan flow cytometry. Adipogenic and osteogenic mediums were used to assess the differentiation ability of the cells. Adenovirus vector deliver green fluorescent protein gene (Ad-GFP) was used to transfected the MSC and the expressing of GFP was detected by fluorescent microscope. The MSC were injected into the liver of newborn rat. The immunofluorescence analysis was conducted to determine the presence of double-positive CD105+/CD166+ cells in different organs of rats. MSC were subcutaneous injected into the human-nonobese diabetes/severe combined immunodeficiency disease (NOD/SCID) mice and carcinogenesises of the MSC in vivo were detected by pathological diagnosis. RESULTS: MSC could be separated from fetal cord blood. These cells were uniformly positive for CD29, CD44, CD59, CD105, CD166 and negative for CD34, CD45, CD80, CD86, HLA-DR. The cells had the abilities to differentiate into adipogenic and osteogenic cells in vitro, expressed the GFP at high levels (56.32%±3.28%). The MSC were located at different organs after injected into the newborn rats and didn't have carcinogenicity in vivo. CONCLUSION: Human second-trimester fetal cord blood MSC is an promising target cells in fetal IUGT.  相似文献   

14.
AIM: To study the effect of natural killer(NK)-cells on graft-versus-host disease(GVHD), graft rejection, engraftment and reconstituting of hematopoiesis in mouse allogeneic bone marrow transplantation. METHODS: Lethally irradiated BALB/c(H-2d) mice were transplanted with C57/6j(H-2b) bone marrow containing donor peripheral T cells and/or NK cells. Recipients CD34+ cell counts and the expression of H-2Kb＋ cell were detected by flow cytometry, peripheral white blood cell(WBC) was detected by auto-cytometry, and the recipients survival rates, GVHD, engraftment and hematopoiesis recovery were then observed. RESULTS: In the group of transplanted with NK cells infusion, the incidence of GVHD was evidently lessened, the survival rates, WBC and CD34+ cell counts and the expression of H-2Kb＋ cell were significantly high than that without NK cells infusion. CONCLUSION: In mouse allogeneic bone marrow transplantation, alloreactivity NK cells prevents GVHD, reduces graft rejection, and promotes engraftment and reconstituting of hematopoiesis.  相似文献   

15.
AIM:To study the effect of TGF-β1 and TNF-α antisense PS-ODNS on ex vivo expansion of hematopoietic stem/progenitor cells (HSPC). METHODS:CD34+cells were purified from fresh umbilical cord blood by immunomagnetic beads, and mononuclear cells were purified from bone marrow by Ficoll-hypaque. The effects of TGF-β1 and /or TNF-α antisense PS-ODNS on ex vivo expansion of CD34+ cells、CFU-GEMM、CFU-GM、CFU-E and BFU-E were detected by using liquid and semi-solid culture systems.RESULTS:TGF-β1 antisense PS-ODNS cooperated with cytokines increased the number of CD34+ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E, which was as 4, 2.6, 2.7, 1.8, 2.1 times as that of the control (the cytokines combination), respectively. TNF-α antisense PS-ODNS cooperated with cytokines respectively increased the number of CD34+ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E by 4, 2.9, 2.6, 1.7, 1.8 times as that of the control. The above two antisense PS-ODNS cooperated with cytokines could respectively increased the number of CD34+ cells, CFU-GEMM, CFU-GM, CFU-E and BFU-E by 5.3, 2.1, 2.7, 1.9, 1.8 times as that of the control.CONCLUSION:Inhibition of endogenous TGF-β1 and TNF-α by antisense PS-ODNS will be one of the effective methods to expand HSPC ex vivo.  相似文献   

16.
AIM: Gene transduction with a recombined murine stem cell retroviral vector has been investigated to find an effective way of gene transduction and to offer theory and experimental basis for the recombined murine stem cell retroviral vector used for gene transduction. METHODS: 1. Construction of retrovirus vector: EC1-4 gene (repeats 1-4 of cadher in-5 extracellular domain) and mutant (Ser 222A) MEK1 gene were cloned into retrovirus vector pMSCV after cut by Bgl Ⅱ and EcoR 1 restriction endonuclease. 2. Obtaining CD41+ cells and cell culture: Cells expressing CD34+ from cord blood were isolated. The inducement of cells expressing CD41 from CD34+ cells was performed by using TPO and cells CD41+ were selected by FACS. NIH 3T3 cells were cultured in high sugar DMEM medium and U937 in RPMI 1640 medium. UT7 cells which is cytokine-dependent cell line were grown in Iscove's modified Dulbeco's medium supplemented by GM-CSF. 3. Determination of viral titers: Retroviral vectors were transferred to packaging cell line 293. Retroviral containing supenatant was collected after transfection. The viral titers was tested on infection of NIH 3T3 cells by FACS analysis. 4. Western blot: Transduced CD41+, UT7, U937 and MDA-MB-435 cells were analyzed by western blot to examine expression of transduced genes. RESULTS: A packaging cell line 293 produces high-titer MEK1 pMSCV retroviruses (3.1×107) and EC1-4 pMSCV retroviruses (1.0×108). With 8-folds dilution retroviruses, 60.73% GFP positive cells have been obtained in MEK1 pMSCV transduced UT7 cells, 72.56% in U937 cells and 30.57% in CD41+ cells, respectively. GFP positive cells have reached up 97.54 % in EC1-4 pMSCV transducted MDA-MB-435 cells. Phosphorylated MEK1 has been decreased in experiment group when TPO has stimulated CD41+ and UT7 cells or serum has stimulated U973 cells. This indicates that is a dominant negative effect of mutation MEK gene. EC1-4 gene transduced MDA-MB-435 cells have expressed EC1-4. CONCLUSION:The recombined murine stem cell retrovirus can effectively mediate gene transduction of CD41+,UT7,U937 and MDA-MB-435 cells,and transduced genes can be stably expressed.  相似文献   

17.
AIM:To establish the method for detecting the immunophenotype of immunosuppressive receptor programmed cell death protein 1 (PD-1) in T-cell receptor (TCR) Vβ repertoire of CD3+, CD4+ and CD8+ T-cell subsets, therefore to evaluate the distribution of PD-1 in T-cell repertoire from human peripheral blood (PB). METHODS:The PB samples from 10 cases of healthy individuals (HI) were collected. Using multi-colored fluorescence flow cytometry, the distribution frequency of PD-1 in TCR Vβ repertoire was detected with a wide panel of anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-PD-1 and 24 anti-TCR Vβ repertoire (IOTest® Beta Mark TCR Vβ Repertoire Kit, containing 8 tubes which labeled A~H, each tube is a composite antibody of FITC and PE coupling, each cocktail contains antibodies direc-ted to 3 different Vβ subfamilies) monoclonal antibodies. RESULTS:The total number of the 24 TCR Vβ repertoire detected in CD3+, CD3+CD4+ and CD3+CD8+ T cells from 10 cases of HI was consistent with the Quick Reference Card data provided by the kit. The preliminary results showed that the frequency of Vβ usage in CD3+, CD4+ and CD8+ T cells was different. High usage of Vβ2, Vβ3, Vβ8, Vβ9, Vβ5.1, Vβ13.1 and Vβ13.2 was found in CD3+ T cells, while high usage of Vβ2, Vβ3, Vβ8, Vβ5.1, Vβ9 and Vβ13.1 in CD3+CD4+ T cells, and high usage of Vβ1, Vβ2, Vβ3, Vβ9, Vβ13.1 and Vβ13.2 in CD3+CD8+ T cells were also observed. Further analysis showed that the expression of PD-1 was detected in all 24 TCR Vβ subfamilies of CD3+, CD3+CD4+ and CD3+CD8+ T cells. The higher frequency of PD-1+ T cells was CD4+Vβ5.2+ T cells, whereas the higher frequency of PD1+ T cells in CD8+Vβ11+ and CD8+Vβ13.6+ T cells was detected. CONCLUSION:The method for detection of the immunosuppressive receptor PD-1 in TCR Vβ repertoire of T-cell subsets is successfully established, which provides a new method for further analysis of immunosuppressive characteristics of TCR Vβ repertoire in the patients with leukemia.  相似文献   

18.
AIM: To investigate the role of mesenchymal stem cell-induced regulatory dendritic cells (MSC-DCregs) in mouse acute graft-versus-host disease(aGVHD) model. METHODS: Bone marrow cells from BALB/c (H-2d) mice were isolated and were induced to differentiate into DCs. The DCs were selected by flow cytometry, and after 10 d co-culture with MSCs, they were induced to be MSC-DCregs. Male 8-week-old C57BL/6 (H-2b) mice were used as donor mice. The female 8-week-old BALB/c (H-2d) mice, who had received 100 cm source-skin distance, 30 cm×30 cm radiation field, 700 cGy total body irradiation (TBI) pretreatment were used as recipient mice. The recipients were divided into 5 groups: control group, TBI group (injected with medium only), bone marrow transplantation group (injected with 1×107 bone marrow cells), aGVHD group (injected with 1×107 bone marrow cells and 1×107 spleen cells), and MSC-DCregs group (injected with 1×107 bone marrow cells, 1×107 spleen cells and 1×106 MSC-DCregs). The white blood cell count, recipients' chimerism, clinical evaluation of aGVHD, survival analysis and pathological changes were determined. RESULTS: Hematopoieic recovery was seen at 10 d after transplantation. The recipients' chimerism was parallel to the donors' at 30 d. The median survival time of the mice in aGVHD group and MSC-DCregs group was 27 d and 33 d, and the survival rates at 30 d were 20% and 100% (P<0.01), respectively. The clinical scores of the mice in MSC-DCregs group were lower than those in aGVHD group (P<0.01). Moreover, the pathological changes in the skin and liver of the mice in MSC-DCregs group were less serious than those in aGVHD group. CONCLUSION: The MSC-DCregs induce an aGVHD tolerance in vivo, and further research of its mechanism is still in great necessary.  相似文献   

19.
AIM: To investigate the heterogeneity of basal intracellular free calcium concentration([Ca2+]i) in peritoneal macrophages(PM) and whether it is relative to the reactivity of PM at the single cell level. METHODS:[Ca2+]iimplicated stimulated were measured by fluorescent microscopic imaging system after loading with fluorescent probe fura-2/AM. Superoxide(O2-)produced by single PM was determined by modified NBT test. RESULTS: The values of basal[Ca2+]idetermined in 392 PMs of 7 mice showed normal distribution [(54±24) nmol/L, n=392] with wide range(less than 20 nmol/L to more than 100 nmol/L), among which about 50% were in the range of 40-60 nmol/L. When stimulated with PMA or fMLP,[Ca2+]iwas increased, the peak values were positively correlated with the basal[Ca2+]iin one mouse(PMA stimulated cells: r=0.52, P<0.01, n=58; fMLP stimulated cells:r=0.59, P<0.01, n=44. Both of the experiments were repeated in 3 mice, the results in the other 2 mice were similar). The O2- in PMA stimulated PMs were also positively correlated with the basal i(r=0.42, P<0.01, n=43, repeated in 4 mice, the results in the other 3 mice were similar). CONCLUSION: Basal[Ca2+]iin murine PM is heterogeneous, and the value of basal[Ca2+]iis tightly correlated to the reactivity of PM stimulated by proinflammatory factors.  相似文献   

20.
AIM: To explore the mechanism of ET-1, NO and PGI2 release from coronary artery endothelial cells(CAEC) induced by acute hypoxia. METHODS: Bovine coronary artery endothelial cells were cultured and [45 Ca2+] was used to investigate the difference of calcium uptake between normoxia group and hypoxia group(3% O2). The contents of ET-1, NO and PGI2 in media of normoxia group, hypoxia group and hypoxia + verapamil group were measured 24 h after hypoxia. RESULTS: [ 45 Ca2+] uptake by CAEC in hypoxia group was 1.9 times more than normoxia group(P< 0.01). Hypoxia + verapamil group released more PGI2, ET-1 and less NO than hypoxia group(P< 0.05). CONCLUSION: Changes of ET-1, NO and PGI2 releases during hypoxia may be caused by the inflow of Ca2+ into coronary artery endothelial cells.  相似文献   

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