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1.
AIM: To investigate the immune stimulation capacity of B7-H1 blockade on immature dendritic cells (DCs) in vitro. METHODS: The human monocyte-derived dendritic cells were induced in the presence of cytokine GM-CSF and IL-4. The expression of B7-H1 was detected by FCM. On blockade of B7-H1, the maturation and endocytic activity, T cells stimulatory proliferation capacity, IL-12 production, T cell differentiation effect of DCs were detected by FCM, MTT assay, ELISA and ELISPOT, respectively. RESULTS: The expression of B7-H1 was increased with the induction of DCs. On day 7, the positive expression was 54.12%, and the TNF-α induced mature DCs had the positive expression rate of 83.64%. The blockade of B7-H1 on immature DCs had sharply increased their T cells stimulatory proliferation capacity and IL-12 production, and efficiently induced the development of Th1/Tc1 cells, but had no effect on their maturation and endocytic activity. CONCLUSION: The blockade of B7-H1 on immature DCs increases its immune stimulation activity. It is valuable to investigate the antitumor immune responses of DCs vaccine with B7-H1 blockade.  相似文献   

2.
AIM: To evaluate the ability of dendritic cells (DCs) loaded with tumor lysate to initiate cell-mediated immune responses by stimulating naive T cells, and the efficiency of activated T cells to kill autologous tumor cells in vitro. METHODS: The peripheral blood lymphocytes and monocytes were obtained from the advanced renal cell carcinoma patient by conglutination method. The immature dendritic cells were generated in the presence of interleukin-4(IL-4) and granulocyte/macrophage colony-stimulating factor(GM-CSF) from monocytes of healthy individuals. These cells were pulsed with tumor lysate or not. Induction of tumor-specific cytotoxic T lymphocytes (CTLs) response by mature dendritic cells (mDCs) was evaluated by the CD95(Fas) expression assay through FCM and the cytotoxic assay against autologous human tumor cells.　RESULTS: Human immature dendritic cells and T cells obtained from healthy donors were stimulated with tumor-pulsed dendritic cells. The immature dendritic cells were applied to the cytotoxicity assay against target autologous tumor cells. The CD95(Fas) expression, IFN-γ and TNF-α secreted by the CTLs in tumor lysate-plused DC group were higher than those of other groups. The capacity of the CTLs to kill autologous tumor cells was significantly different(P<0.05). Antigen-specific DCs vaccine can induce T cells activation and proliferation, thus we can obtain higher proportion of tumor specific cytotoxic T cells(CTLs), and enhance the CTLs to secret IFN-γ and TNF-α. CONCLUSION: Our results indicate that monocyte-derived human dendritic cells pulsed with tumor lysate could induce the specific antitumor effect against autologous tumors . This in vitro model offers a new and simple approach to the development of DC+CTL-based immunotherapy.  相似文献   

3.
AIM: To study the effects of progesterone (P4) on the maturation and immunologic function of dendritic cells (DCs) from human peripheral blood. METHODS: Cultured DCs were treated with P4 at doses of 10-7 mol/L and 10-6 mol/L. The morphologic changes were observed under the scanning electronic microscope. The immunophenotypes of DCs in control and treated groups were analyzed by flow cytometry. IL-10 and IL-12 production in culture supernatant was examined by ELISA assay. The capability of the stimulatory activity of the DCs on allogeneic T cells in mixed reaction was tested by incorporation of ［3H］-TdR. RESULTS: Compared with control group, cultured DCs in the presence of P4 displayed less dendritic pseudopod, expressed low levels of MHC-II, CD40, CD80 and CD86, and exhibited weakly activity in stimulating the proliferation of allogeneic T cells. Increase in IL-10 production and decrease in IL-12 production were observed. CONCLUSION: P4 exerts negative effect on the maturation and immunologic function in dendritic cells from human peripheral blood.  相似文献   

4.
AIM: To investigate the effects of 17β-estradiol (E2) on the maturation and immunologic function of dendritic cells from human peripheral blood. METHODS: Cultured DCs were treated with E2 at doses of 10-7 mol/L and 10-6 mol/L. The morphologic changes were observed under the scanning electronic microscope. The immunophenotype of DCs in control and treated groups was analyzed by flowcytometry. IL-12 production in culture supernatant was examined by ELISA assay. The capability of the stimulatory activity of the DCs on allogeneic T cells in mixed reaction was tested by incorporation of ［3H］-TdR. RESULTS: Compared with control group, DCs cultured in the presence of E2 displayed less dendritic pseudopod, expressed low levels of MHC-II, CD40, CD80 and CD86, and exhibited weakly activity in stimulating the proliferation of allogeneic T cells and reduction of IL-12 production. CONCLUSION: E2 exerts a negative effect on the maturation and immunologic function of dendritic cells from human peripheral blood.  相似文献   

5.
AIM: To investigate the immunological effect of tumor necrosis factor receptor-associated factor 6 (TRAF6) on the maturation of dendritic cells in vitro.METHODS: The human peripheral blood mononuclear cell (PBMC)-derived dendritic cells (DCs), induced by recombined human granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4, were stimulated to mature with TNF-α, LPS or cocktail of cytokines (TNF-α, IL-6, IL-1β and PGE2).The cell surface markers, T-cell stimulatory proliferation capacity and IL-12 p40 production of the DCs were determined by the methods of flow cytometry, ELISA and mixed lymphocyte reation (MLR), respectively.The mRNA expression of TRAF6 was detected by real-time PCR.RESULTS: The expression of TRAF6 was observed in all groups, which in cocktail group was the highest in the DCs with optimum state of maturation.Furthermore, TRAF6 enhanced the production of IL-12 and the ability of T-cell stimulation of mature DCs.CONCLUSION: TRAF6 might play an important role in inducing the maturation of human PBMC-derived DCs.  相似文献   

6.
AIM: To study the dynamic levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and gamma interferon （IFN-γ） produced by human dendritic cells infected with Dengue virus. METHODS: Monocytes isolated from healthy human peripheral blood were incubated in medium with GM-CSF and IL-4 for more than 7 days. DCs were then collected and identified by scanning electron microscope, immunohistochemistry and lymphocytes stimulatory ability. Human dendritic cells （DC) were infected with Dengue-2 virus (DV-2) in vitro， culture supernatants were collected in different time postinfection (6 h, 12 h, 24 h, 48 h and 72 h)， DV antigen in human dendritic cells were demonstrated by an indirect immunofluorescent assay （IFA）, production of TNF-α, IL-6 and IFN-γ in the culture supernatants were evaluated by ELISA. RESULTS: After 7 days, typical dendritic cells could be obtained. Virus antigen were detected in infected DC by IFA. Dengue virus induces TNF-α and IL-6 secretion from DC and does not induce IFN secretion. CONCLUSION: Human dendritic cells are target of dengue virus infection. TNF-α， IL-6 production from DC are increased with DV infection. Dendritic cells may play an important role in DV pathogenicity and immunity.  相似文献   

7.
AIM: To study the effect of interleukin 18 gene transfected lung cancer cells on the phenotype and immunological activity of dendritic cells (DC). METHODS: A secretive IL-18 expression vector containing IL-12 P40 signal sequence was constructed and transfected into NCI-H460 lung cancer cells. DC induced from human peripheral blood were divided into 4 groups (NT, PV, GT and PD). DC were stimulated by non-transfected NCI-H460 cells, pure vector transfected NCI-H460 cells and IL-18 transfected NCI-H460 cells respectively for group NT, PV, GT, and non-stimulated DC for group PD. CD54, CD80, CD83 and CD86 on DC in the 4 groups were detected with flow cytometry. T cell proliferation stimulated by DC in the 4 groups was assayed with MTT method. IL-12 release in cultured DC supernatant was measured by ELISA. RESULTS: Sequencing result of the secretive IL-18 was correct. The transfected cells expressed IL-18 fusion gene and 18 kD IL-18 protein. DC in GT group expressed more surface molecules than those in other 3 groups. T cell proliferation and IL-12 secretion in GT group were higher than those in other 3 groups. CONCLUSION: IL-18 gene transfected NCI-H460 cell increases surface molecule expressions on DC. It also enhances immunological activity and IL-12 secretion in DC.  相似文献   

8.
AIM: To investigate the immune function of dendritic cells (DCs) in patients with hepatocellular carcinoma(HCC). METHODS: The DCs were cultured from human peripheral blood mononuclear cells (PBMC) by using GM-CSF and IL-4 for 7 days. Surface molecules CD86, HLA-DR of DCs were detected by flow cytometry. IL-12 production by DCs and IFN-γ production by T cells was measured with ELISA and ELISPOT, respectively. Allogenic mixed lymphocyte reaction was detected by MTT assay. RESULTS: CD86 expression in DCs in HCC patients were markedly lower than that in health control (91.7％ vs 83.5%, P<0.05). IL-12 production by DCs had no significant difference between the HCC patients and the health control ［(324.6±171.0)ng/L vs (436.5±142.7)ng/L, P>0.05］. However, after stimulated DCs with LPS, IL-12 production in HCC patients was significantly lower than that in health control ［(478.6±142.7)ng/L vs (630.0±151.9)ng/L, P<0.05］. IFN-γ production by T cells and T cell proliferation index (PI) in the HCC patients were all significantly lower than those in health control ［(IFN-γ: 133.4±51.2)103U/L vs (183.0±60.2)103U/L, P<0.05; PI: 2.3±0.7 vs 3.5±0.8, P<0.01］. CTL killing activity assay indicated that DC-induced CTL killing activity in HCC group was significantly lower than that in health group when the E/T ratio was 10∶1 and 20∶1, respectively. CONCLUSION: The immune function of DCs and T cells in the patients with HCC are significantly decreased. The CTL killing activity of HCC group is also markedly decreased.  相似文献   

9.
WANG Ling  LUO He-sheng  XIA Hong 《园艺学报》2009,25(10):1950-1953
AIM: Sodium butyrate has antitumor effects on colon cancer cells such as inhibiting cell growth and promoting differentiation and apoptosis. The aim of this study is to investigate whether sodium butyrate induces apoptosis in human colon cancer cell line HT-29 and to examine the intracellular mechanisms involved, especially the role of caspase activation in the process. METHODS: HT-29 cells were cultured to logarithmic phase before treatment with sodium butyrate at concentration of 5.0 mmol/L and caspase inhibitors at the concentration of 20 μmol/L. The latter were added in the medium ahead of sodium butyrate for 1 h. Then, the staining of Annexin V-FITC and PI were used to analyze HT-29 apoptosis and the dye JC-1 was applied to detect mitochondrial membrane potential by flow cytometry. Caspase activity within the cells was measured respectively using a specific caspase activity assay kit and a microplate reader. RESULTS: Preincubation of HT-29 cells with sodium butyrate significantly increased apoptosis ［(35.40±0.70)%］ and decreased mitochondrial membrane potential (5.53±0.91). This effect was blocked when pretreatments were enforced with z-VAD-fmk, z-DEVD-fmk and z-LEHD-fmk. The apoptosis percentages were (1.33±0.59)%, (1.40±0.53)% and (1.27±0.91)%, respectively and mitochondrial membrane potentials were 9.80±1.15, 10.23±0.50 and 10.33±1.02, respectively. However, the role of reduction by z-IETD-fmk, which presented the apoptosis percentage of (32.10±2.33)% and mitochondrial membrane potential of 5.93±1.31, was not observed. An enhancement of caspase-3 and -9 activities (2-3-fold) but no change of caspase-8 activity was confirmed. CONCLUSION: Apoptosis of HT-29 colon carcinoma cells induced by sodium butyrate is tightly linked to caspase activation via mitochondrial pathway other than tumor necrosis factor-alpha and has the potential to inhibit proliferation and thereby may contribute to the progression of colon cancer.  相似文献   

10.
WANG Yi  HAO Yu  LOU Jin-li  MA Hui  QIU Quan-ying 《园艺学报》2004,20(10):1759-1764
AIM: To study the effect of ginsenoside Rg1 and Rh1 on the anti-tumor activity of dendritic cells (DC). METHODS: Effect of Rg1 and Rh1 on the production of IL-12 p40 protein was detected by ELISA, and the IL-12 p40 mRNA level of DC was monitored by RT-PCR. Anti-tumor activity of DC-LPAK was determined by neutral red staining assay. RESULTS: The results of ELISA showed that Rg1 and Rh1 significantly enhanced the production of IL-12 p40 of DC. Rg1 at 1 mg/L and Rh1 at 100 mg/L upregulated the IL-12 p40 mRNA level. Rg1 and Rh1 enhanced the anti-tumor ability of DC, induced lymphokine and PHA activated killer (DC- LPAK) on human papillate tumor cell line. Each dose of Rg1 can obviously accelerate the cytotoxity to L929 at the E∶T ratio of 5∶1(P<0.05,0.01), while only Rh1 10 mg/L enhanced the cytotoxity ability of DC-LPAK (P<0.05). CONCLUSION: Rg1 and Rh1 enhanced the production of IL-12 p40. This effect may be mediated by the increase in the mRNA level. As a result, Rg1 and Rh1 promote the ability of DC to stimulate the cytotoxitic acticity of DC-LPAK.  相似文献   

11.
AIM: To investigate the effect of interleukin-13 (IL-13) on interleukin-12 (IL-12) production in mesangial cells.METHODS: The protein synthesis of IL-12 in mesangial cells was measured by ELISA.The expression of IL-12 mRNA in mesangial cells was evaluated by RT-PCR.RESULTS: The production of IL-12 in mesangial cells stimulated by lipopolysaccharide(LPS) was significantly increased (P<0.01).IL-13 (1-100 μg/L) inhibited the protein and mRNA expression of IL-12 in a dose-dependent manner (P<0.05 or P<0.01).CONCLUSION: IL-13 inhibits IL-12 expression induced by LPS in mesangial cells.IL-13 may regulate immune responses by balancing Th1/Th2 in glomerulonephritis.  相似文献   

12.
AIM: To investigate the effect of uric acid on the maturation and the biological function of murine bone-marrow derived dendritic cells (BMDCs) in vitro.METHODS: BMDCs were cultured with GM-CSF, IL-4, LPS and uric acid. Cell phenotypes were analyzed by flow cytometry. MTT was used to detect the effect of uric acid on the function of BMDCs in stimulating the proliferation of T cells. IL-12 released by BMDCs was also detected. RESULTS: BMDCs were cultured and identified. Uric acid at concentrations of 200 mg/L and 400 mg/L increased the expression of the molecules (CD11c, CD83, CD86, IA/IE) on BMDCs surface and the IL-12 level in the culture supernatants (P<0.05), promoted the proliferation of T cells at the T: DC rate 5∶1, 10∶1, 20∶1 (P<0.05). However, uric acid at concentration of 70 mg/L had no effect on above molecule expression (P>0.05), no effect on T cell proliferation with BMDCs (P>0.05) was observed. CONCLUSION: Uric acid promotes the differentiation, maturation, the expression of co-stimulatory molecules, the IL-12 production in BMDCs and enhances the ability of BMDCs to stimulate the proliferation of T cell in a specical dose range.  相似文献   

13.
AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-κB activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P＜0.05). The data of mixed leukocyte reaction and IL-12P70 production were consistent with the flow cytometry results (P＜0.01). Nuclear extracts from the anti-CD47 mAb-treated DCs revealed a decrease in NF-κB binding activity. CONCLUSION: The anti-CD47 mAb exerts a negative effect on the maturation and function of in vitro cultured DCs via inhibiting of NF-κB binding activity.  相似文献   

14.
AIM: To study the effects of WT1 peptide-loaded dendritic cells (DC) stimulating the cytotoxic T lymphocytes (CTL) on K562 cells in vitro. METHODS: DC were generated from normal human peripheral blood mononuclear cells (PBMC) in the presence of granulocyte-macrophage colony stimulating factor(GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-α) , DC were cultured with WT1 peptides , and then triggered T cells into specific CTL. RESULTS: Most suspended cells exhibited distinctive morphological features of DCs and they stimulated proliferation of allogenic lymphocytes. Under the effector : target ratio of 20∶1, CTLs derived from cultures with DC and WT1 peptides were showed 86.1%±26.8% cytotoxicity against K562 cells, cytotoxicity by CTLs derived from cultures with unloaded DC against K562 cells were 47.1%±20.8% and cytotoxicity by lymphocytes were 27.7%±15.3%. Cytotoxicity by CTLs derived from culture with WT1 peptides-loaded DC were the strongest among three groups （P＜0.05）. CONCLUSION: CTLs derived from cultures containing DC pulsed with WT1 peptides show the strongest cytolytic activities on K562 cells.  相似文献   

15.
AIM:To investigate the relation of tolerogenic dendritic cells (DC) induced by interleukin-10 (IL-10) and the paired immunoglobin-like receptor (PIR) A and B (PIR-A and PIR-B) in mouse. METHODS:The mouse dendritic cell line, DC2.4 cells were cultured with the IL-10 to develope the IL-10-DC and were stimulated by lipopolysaccharide (LPS) for 48 h to induce the mature dendritic cells (LPS-DC). Special small inference RNA (siRNA) molecule of PIR-B was chemically synthesized and was transfected into IL-10-DC by Lipofectamine 2000 (Si-DC). The expression of PIR A and PIR B on DC2.4 cells were measured by semi-quantitative RT-PCR and flow cytometry (FCM). The allogeneic stimulating capacity of DC was measured by mixed lymphocyte reaction (MLR) using ［3H］-thymidine incorporation test. The concentration of IFN-γ in supernatants of MLR from distinct groups was analyzed by ELISA. RESULTS:IL-10 up-regulated the PIR-B and down-regulated the PIR-A by semi-quantitative RT-PCR. On the contrary, LPS down-regulated the PIR-B expression and up-regulated the PIR-A expression. The expression of PIR, which is the common extra-membrane of PIR-A and PIR-B, was increased in both the IL-10-DC and the LPS-DC groups by FCM detection, but the higher expression was found in IL-10-DC group than that in LPS group. The IL-10 induced the higher PIR-B expression, inhibited allogenetic T cell proliferation and down-regulated the IFN-γ secretion. Special siRNA molecules of PIR-B in IL-10 group promoted the T cell proliferation and enhanced the IFN-γ secretion in MLR. CONCLUSION:IL-10 up-regulates the PIR-B expression and makes DC tolerance. Up-regulated PIR-B expression may be the molecular mechanism of tolerogenic dendritic cells induced by IL-10 in mouse.  相似文献   

16.
AIM and METHODS:To study the immunological effect of measles vaccine therapy on asthmatic children, we examined the changes of interleukin-12 , interleukin-13 and total serum IgE levels in plasma and cultured peripheral blood mononuclear cells(PBMC) supernatant by means of ELISA in 13 mild-moderate asthmatic children treated with measles vaccine. Results were compared with 12 anti-symptomatic treatment mild-moderate asthmatic children and 17 normal children control group.RESULTS:After measles vaccine treatment, IL-13 and total serum IgE levels decreased remarkably, statistically lower than that of group receiving only anti-symptomatic treatment. There was no statistical difference in IL-12 level between the two group. Correlation analysis: 1)IL-12 level of plasma was negatively correlated to the level of serum total IgE, there was no correlation of supernatant IL-12 in PBMC to the total serum IgE; 2)IL-13 levels in plasma and PBMC were positively correlated to the level of total serum IgE; 3) IL-12 level was negatively correlated to IL-13.CONCLUSION:Measles vaccine could down-regulate IL-13 level,therefore decrease total IgE synthesis,but not affect IL-12 level in asthmatic children.  相似文献   

17.
AIM: To investigate the phenotype and immune activity of dendritic cells using interleukin-18 as intervent.METHODS: Monocytes were isolated from human peripheral blood and induced into DCs with GM-CSF and IL-4. The cellular morphous was observed under inverted microscope. On the 5th day, 3 groups including IL-18 group, TNF-α group and IL-18+TNF-α group were set. IL-18, TNF-α or IL-18+TNF-α was used as intervents respectively to facilitate cell maturity. Supernatants were collected at 24 h, 48 h and 72 h. IL-12 in the supernatant, CD1a, HLA-DR, CD83 and CD86 were analyzed using flow cytometry. DCs of the 3 groups were co-cultured with T cells respectively on the ratio of 1∶〖KG-*2〗100, 1∶〖KG-*2〗50 and 1∶〖KG-*2〗10. T cell proliferation stimulated by DC was determined using MTT method. DCs were co-cultured with T cells on the ratio of 1∶〖KG-*2〗10, and the supernatant were collected at 24 h, 48 h and 72 h. IFN-γ in the supernatant was detected with ELISA method.RESULTS: Induced by GM-CSF and IL-4, then stimulated by IL-18, TNF-α or IL-18+TNF-α, monocytes showed typical morphous of DC. No morphological difference was observed among DCs of the 3 groups. No statistical difference showed in expression level of CD1a, HLA-DR, CD83 and CD86 between IL-18 group and TNF-α group (P>0.05). The positive rates of CD1a and CD83 in IL-18+TNF-α group were higher than those in other 2 groups. The positive rate of HLA-DR in IL-18+TNF-α group was higher than that in IL-18 group. No difference between IL-18 group and TNF-α group in the potency of stimulating T cell proliferation was found, whereas the stimulating potency in IL-18+TNF-α group was higher than that in IL-18 group and TNF-α group. IL-12 in IL-18+TNF-α group at 48 h and 72 h was higher than that in IL-18 group and TNF-α group (P<0.05). However, there was no difference between the latter 2 groups. There was also no difference between IL-18 group and TNF-α group in IFN-γ secretion. IFN-γ in IL-18+TNF-α group was higher than that in IL-18 group and TNF-α group (P<0.05).CONCLUSION: Using IL-18 as intervent, DC expresses high level of surface molecules, secretes high level of IL-12, stimulates T cell proliferating effectively and produces IFN-γ potently. The actions are stronger when used in combination with TNF-α. It suggests that IL-18 may serve as a promoting agent of DC maturity, or combination with TNF-α in DC induction will strengthen the immune activity of DC.  相似文献   

18.
AIM: To investigate the effects of transforming growth factor β1 (TGF-β1) on murine-derived dendritic cells (DC). METHODS: Murine bone marrow cells were cultured with GM-CSF and TGF-β1 to develop TGF β-DC. Then they were stimulated by lipopolysaccharide (LPS). Their phenotypes were assessed by flow cytometry (FCM). The allogeneic stimulating capacity of DC was measured by mixed lymphocyte reaction (MLR) using BrdU ELISA method. IL-12 p70 protein was detected by ELISA and the expressions of Toll like receptor 4 (TLR4) on DCs were measured by semi-quantitative RT-PCR and FCM. RESULTS: Compared to immature DC (imDC) cultured with GM-CSF alone, the expressions of CD80, CD86, I-Ab and CD40 in TGF β-DC were lower. The TGF β-DC was resistant to maturation by LPS. Maturation resistance was evident from a failure to up-regulate CMs, to stimulate larger T cell proliferation and to increase secretion of IL-12 p70. Down-regulation of TLR4 expression on TGF β-DC was also found. CONCLUSION: TGF-β1 inhibits the expression of co-stimulatory molecules on DC. It is resistant to maturation stimulus (LPS) and might be linked with TLR4 down-regulation.  相似文献   

19.
AIM: To observe the effect of B7H1 expression in pancreatic carcinoma cells on the proliferation and activation of co-cultured T lymphocytes. METHODS: B7H1 expression in panc-1 cells before and after interferon-γ(IFN-γ) treatment or B7H1-siRNA transfection was evaluated by RT-PCR and flow cytometry. The influence of B7H1 expression on co-cultured PHA-activated T lymphocytes was determined by the methods of MTT and enzyme-linked immunosorbent assay (ELISA). RESULTS: B7H1 was highly expressed in panc-1 cells and up-regulated after IFN-γ stimulation. Such up-regulation led to the significant inhibition of T cell proliferation and secretion of cytokines such as IFN-γ and interleukin-2(IL-2). However, IL-10 production was enhanced. In contrast, knockdown of B7H1 expression in panc-1 cells by RNA interference resulted in increased T cell proliferation as well as IFN-γ and IL-2 production. Meanwhile, the IL-10 secretion decreased. CONCLUSION: B7H1-expressing panc-1 cells suppress T cell function by inhibiting T cell proliferation and production of Th1 cytokines. Suppression of B7H1 expression through siRNA restores T cell immune functions, indicating a potential strategy for immunotherapy against pancreatic cancer.  相似文献   

20.
AIM: The goal of this study was to compare different methods for tumor antigen preparation, to observe the induction of tumor-specific cytotoxic T lymphocytes in rats by dendritic cells (DCs) pulsed with different tumor antigens. METHODS: The precursors of dendritic cells were isolated from bone marrow of rats, stimulated in vitro with recombinent rat granulocyte-macrophage colony-stimulating factor (rrGM-CSF) and interleukin-4 (rrIL-4). Then rat DCs were pulsed with C6 tumor cell antigens prepared with different methods: freeze-thaw, boiling or total protein extracted from ultrasonic crushed tumor cell. Subsequently primed DCs were cocultured with T lymphocytes isolated from spleen to induce CTL. Lymphocyte chemoattractant factor from DCs and cytokine IFN-γ release were determined by ELISA, the cytotoxicity of CTL was assayed by JAM test. RESULTS: DCs pulsed with boiled tumor cell in vitro induced an enhanced ability of T-cell proliferation and cytotoxic T lymphocyte activity.CONCLUSION: Our results demonstrated that DCs primed with boiled tumor cell may represent a method for inducing immune responses against the entire repertoire of tumor antigens of malignancies.  相似文献   

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