Objective:CD8+T cells are the key effector cells in the anti-tumor immune response.The mechanism underlying the infiltration of CD8+T cells in esophageal squamous cell carcinoma(ESCC)has not been clearly elucidated.Me...Objective:CD8+T cells are the key effector cells in the anti-tumor immune response.The mechanism underlying the infiltration of CD8+T cells in esophageal squamous cell carcinoma(ESCC)has not been clearly elucidated.Methods:Fresh ESCC tissues were collected and grouped according to the infiltration density of CD8+T cells.After the transcriptome sequencing on these samples and the combined analyses with The Cancer Genome Atlas(TCGA)ESCC data,a secreted protein DEFB1 was selected to explore its potential role in the infiltration of CD8+T cells.Bioinformatics analyses,histological verification and in vitro experiments were then performed.Results:DEFB1 was highly expressed in ESCC,and the high expression of DEFB1 was an independent risk factor for overall survival.Since the up-regulation or down-regulation of DEFB1 did not affect the proliferation,migration and apoptosis of ESCC cells,we speculated that the oncogenic effect of DEFB1 was achieved by regulating microenvironmental characteristics.Bioinformatics analyses suggested that DEFB1 might play a major role in the inflammatory response and anti-tumor immune response,and correlate to the infiltration of immature dendritic cell(imDC)in ESCC.Histological analyses further confirmed that there were less CD8+T cells infiltrated,less CD83+mature DC(mDC)infiltrated and more CD1a+imDC infiltrated in those ESCC samples with high expression of DEFB1.After the treatment with recombinant DEFB1 protein,the maturation of DC was hindered significantly,followed by the impairment of the killing effects of T cells in both 2D and 3D culture in vitro.Conclusions:Tumor-derived DEFB1 can inhibit the maturation of DC and weaken the function of CD8+T cells,accounting for the immune tolerance in ESCC.The role of DEFB1 in ESCC deserves further exploration.展开更多
AIM:To investigate possible differences in dendritic cells(DC)within intestinal tissue of mice before and after induction of colitis. METHODS:Mucosal DC derived from intestinal tissue,as well as from mesenteric lymph ...AIM:To investigate possible differences in dendritic cells(DC)within intestinal tissue of mice before and after induction of colitis. METHODS:Mucosal DC derived from intestinal tissue,as well as from mesenteric lymph nodes and spleen,were analyzed by fluorescence activated cell sorting(FACS) analysis.Supernatants of these cells were analyzed for secretion of different pro-and anti-inflammatory cytokines. Immunohistochemistry and immunofluorescence were performed on cryosections of mucosal tissue derived from animals with colitis as well as from healthy mice. RESULTS:It was shown that DC derived from healthy intestinal lamina propria(LP)represented an immature phenotype as characterized by low-level expression of costimulatory cytokines.In contrast to DC from spleen and mesenteric lymph nodes(MLN)that secreted proinflammatory cytokines,LP-DC produced high levels of the anti-inflammatory cytokine IL-10.After induction of mu-rine colitis in a CD4 + CD62L + transfer model or in chronic Dextran sulfate sodium-colitis,a marked increase of activated CD80 + DC could be observed within the inflamed colonic tissue.Interestingly,in contrast to splenic DC,a significant population of DC within MLN and colonic LP expressed the mucosal integrin CD103 which was lost during colitis. CONCLUSION:The constitutive secretion of antiinflammatory cytokines by immature DC within the intestinal LP might regulate the homeostatic balance between mucosal immunity and tolerance.CD103 + DC could mediate this important function.展开更多
BACKGROUND Blastic plasmacytoid dendritic cell tumor(BPDCN)is a rare and highly invasive lymphohematopoietic tumor that originates from plasmacytoid dendritic cells.BPDCN has an extremely poor prognosis.Skin lesions a...BACKGROUND Blastic plasmacytoid dendritic cell tumor(BPDCN)is a rare and highly invasive lymphohematopoietic tumor that originates from plasmacytoid dendritic cells.BPDCN has an extremely poor prognosis.Skin lesions are usually the first manifestation of BPDCN,although the tumor may also invade the bone marrow,lymph nodes,peripheral blood,and other parts of the body,leading to several other manifestations,requiring further differentiation through skin biopsy and immunohistochemistry.CASE SUMMARY In the present paper,the cases of 2 patients diagnosed with BPDCN are discussed.The immunohistochemistry analysis of these 2 patients revealed positivity for CD4,CD56,and CD123.Currently,no standard chemotherapy regimen is available for BPDCN.Therefore,intensive therapy for acute lymphoblastic leukemia was applied as the treatment method for these 2 cases.CONCLUSION Although allogeneic bone marrow transplantation could be further effective in prolonging the median survival the ultimate prognosis was unfavorable.Future treatment modalities tailored for elderly patients will help prolong survival.展开更多
The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These syst...The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These systems can induce specific cell reactions,promote specific tissue functions,and serve as valuable tools for research in tissue engineering,regenerative medicine,and drug discovery.This paper discusses current developments in the field of three-dimensional cell culture and the potential applications of 3D type 1 collagen gels to enhance the growth and maturation of dendritic cells.展开更多
Objective: To observe the lethal effect of T cells activated by dendritic cells (DCs) with MUC1 gene transfection (MUC1-DCs) on BIU-87 cells in vitro, and to evaluate the possibility of dendritic cells transfecte...Objective: To observe the lethal effect of T cells activated by dendritic cells (DCs) with MUC1 gene transfection (MUC1-DCs) on BIU-87 cells in vitro, and to evaluate the possibility of dendritic cells transfected by MUC1 gene as a vaccine for bladder cancer immunotherapy. Methods: MUC1 was successfully transfected into cultured human blood-derived dendritic cell with lipofectin. The transfection efficiency and immunocompetence were detected by flow cytometry and MTT colourmetry. T lymphocytes activated by MUC1-DCs were used to kill BIU-87 cell lines and normal bladder epithelium in vitro and the killing rate was evaluated by MTT colourmetry. Results: There was significant lethal effect on the BIU-87 cells and normal bladder epithelium with T cells activated by MUCl-transfected DCs, but the lethal effect on the BIU-87 cells significantly exceeded that on normal bladder epithelium (P〈0.05). The lethal effect on BIU-87 cells with T cells activated by MUCl-transfected DCs significantly exceeded that with T cells activated by no-transfection DCs (P〈0.01). Conclusion: T cells activated by MUC1-DCs could induce killing action to BIU-87 cell lines. MUC1 gene could be used as a target for bladder cancer immunotherapy.展开更多
AIM:To investigate the function of PU.1-silenced semimature dendritic cells(DCs)and the possibility of utilizing cell immunity in rat intestinal transplantation.METHODS:DCs were isolated from the bone marrow of F344 r...AIM:To investigate the function of PU.1-silenced semimature dendritic cells(DCs)and the possibility of utilizing cell immunity in rat intestinal transplantation.METHODS:DCs were isolated from the bone marrow of F344 rats and cultured using the adherent method.The PU.1 gene was knocked down in DCs using small interfering RNAs(siRNAs)for 24 h,and the cells were then incubated with lipopolysaccharide for 48 h.The PU.1 siRNA that had the highest silencing efficiency was screened using reverse transcription-polymerase chain reaction and Western blot for further study.The tolerance capacity was analyzed and compared between PU.1-silenced DCs(siRNA PU.1 group),negative control-silenced DCs(siRNA NC group)and immature DCs(control group)both in vitro and in vivo.CONCLUSION:Blocking expression of the PU.1 gene in vitro led to a reduction in DC maturation and an increased tolerance capability.PU.1-silenced DCs expressed moderate levels of major histocompatibility complex(MHC)-Ⅱand low levels of co-stimulatory molecules,and produced more interleukin(IL)-10,but less IL-12.Compared with the negative control group,surface molecules cluster of differentiation 80(CD80),CD86 and MHC-Ⅱin the siRNA PU.1 group were 27.0%±5.6%,23.6%±4.8%and 36.8%±6.8%,respectively,and showed a significantly lower trend(P<0.05).In vivo treatment of recipients with PU.1-silenced DCs injected before intestinal transplantation(siRNA PU.1group),significantly prolonged allograft survival and resulted in better tissue histopathology compared with the siRNA NC group and control group.Mean survival time after transplantation was 14.3±3.3 d in the siRNA PU.1 group(P<0.05).CONCLUSION:PU.1-silenced semi-mature DCs induced partial immune tolerance both in vitro and in vivo,which could be used as a new strategy to promote transplantation tolerance.展开更多
Type 1 diabetes(T1D)is an autoimmune disease characterized by the destruction of insulin-producingβ-cells of the pancreatic islets by autoreactive T cells,leading to high blood glucose levels and severe long-term com...Type 1 diabetes(T1D)is an autoimmune disease characterized by the destruction of insulin-producingβ-cells of the pancreatic islets by autoreactive T cells,leading to high blood glucose levels and severe long-term complications.The typical treatment indicated in T1D is exogenous insulin administration,which controls glucose levels;however,it does not stop the autoimmune process.Various strategies have been implemented aimed at stoppingβ-cell destruction,such as cellular therapy.Dendritic cells(DCs)as an alternative in cellular therapy have gained great interest for autoimmune disease therapy due to their plasticity to acquire immunoregulatory properties both in vivo and in vitro,performing functions such as anti-inflammatory cytokine secretion and suppression of autoreactive lymphocytes,which are dependent of their tolerogenic phenotype,displayed by features such as semimature phenotype,low surface expression of stimulatory molecules to prime T cells,as well as the elevated expression of inhibitory markers.DCs may be obtained and propagated easily in optimal amounts from peripheral blood or bone marrow precursors,such as monocytes or hematopoietic stem cells,respectively;therefore,various protocols have been established for tolerogenic(tol)DCs manufacturing for therapeutic research in the treatment of T1D.In this review,we address the current advances in the use of tolDCs for T1D therapy,encompassing protocols for their manufacturing,the data obtained from preclinical studies carried out,and the status of clinical research evaluating the safety,feasibility,and effectiveness of tolDCs.展开更多
Objective: To explore the effects of transforming growth factor (51 (TGF-β1) on dendritic cells (DC). Methods: Murine bone marrow cells were cultured with GM-CSF and TGF-β1 to develop TGF-β1-treated DC (TGFβ-DC). ...Objective: To explore the effects of transforming growth factor (51 (TGF-β1) on dendritic cells (DC). Methods: Murine bone marrow cells were cultured with GM-CSF and TGF-β1 to develop TGF-β1-treated DC (TGFβ-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 and IL-12p70 protein was detected by ELISA. The expression of Toll-like receptor 4 (TLR4) was analyzed by semi quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and FCM. Results: Compared to immature DC (imDC) cultured by GM-CSF alone, the TGFβ-DC express lower CD80, CD86,I-Ab and CD40. The TGFβ-DC were resistant to maturation with LPS. Maturation resistance was evident from a failure to up-regulate co-stimulatory molecules (CMs), to stimulate larger T cells proliferation and to enhance secretion of IL-12p70. We also found that TGF-β1 could down-regulate TLR4 expression on TGFβ-DC. Conclusion: TGFβ-DC are resistant to maturation stimulus (LPS) and might have some correlation with the down-modulation of TLR4 expression.展开更多
BACKGROUND Enterotoxigenic Bacteroides fragilis(ETBF)causes colitis and diarrhea,and is considered a candidate pathogen in inflammatory bowel diseases as well as colorectal cancers.These diseases are dependent on ETBF...BACKGROUND Enterotoxigenic Bacteroides fragilis(ETBF)causes colitis and diarrhea,and is considered a candidate pathogen in inflammatory bowel diseases as well as colorectal cancers.These diseases are dependent on ETBF-secreted toxin(BFT).Dendritic cells(DCs)play an important role in directing the nature of adaptive immune responses to bacterial infection and heme oxygenase-1(HO-1)is involved in the regulation of DC function.AIM To investigate the role of BFT in HO-1 expression in DCs.METHODS Murine DCs were generated from specific pathogen-free C57BL/6 and Nrf2−/−knockout mice.DCs were exposed to BFT,after which HO-1 expression and the related signaling factor activation were measured by quantitative RT-PCR,EMSA,fluorescent microscopy,immunoblot,and ELISA.RESULTS HO-1 expression was upregulated in DCs stimulated with BFT.Although BFT activated transcription factors such as NF-κB,AP-1,and Nrf2,activation of NF-κB and AP-1 was not involved in the induction of HO-1 expression in BFT-exposed DCs.Instead,upregulation of HO-1 expression was dependent on Nrf2 activation in DCs.Moreover,HO-1 expression via Nrf2 in DCs was regulated by mitogenactivated protein kinases such as ERK and p38.Furthermore,BFT enhanced the production of reactive oxygen species(ROS)and inhibition of ROS production resulted in a significant decrease of phospho-ERK,phospho-p38,Nrf2,and HO-1 CONCLUSION These results suggest that signaling pathways involving ROS-mediated ERK and p38 mitogen-activated protein kinases-Nrf2 activation in DCs are required for HO-1 induction during exposure to ETBF-produced BFT.展开更多
Heat shock proteins (HSPs) are reported to act as effective adjuvants to elicit anti-tumor and anti-infection immunity. Here, we report that Hsp70-like protein 1 (Hsp70L1), a novel HSP derived from human dendritic cel...Heat shock proteins (HSPs) are reported to act as effective adjuvants to elicit anti-tumor and anti-infection immunity. Here, we report that Hsp70-like protein 1 (Hsp70L1), a novel HSP derived from human dendritic cells (DCs), has potent adjuvant effects that polarize responses toward Th1. With a calculated molecular weight of 54.8 kDa, Hsp70L1 is smaller in size than Hsp70 but resembles it both structurally and functionally. Hsp70L1 shares common receptors on DCs with Hsp70 and can interact with DCs, promoting DC maturation and stimulating secretion of the proinflammatory cytokines interleukin 12p70 (IL-12p70), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), and the chemokines IP-10, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and normal T cell expressed and secreted (RANTES). The induction of interferon-gamma-inducible protein 10 (IP-10) secretion by Hsp70L1 is not shared by Hsp70, and other functional differences include more potent stimulation of DC IL-12p70, CC-chemokine, and CCR7 and CXCR4 expression by Hsp70L1. Immunization of mice with the hybrid peptide Hsp70L1-ovalbumin(OVA)(257-264) induces an OVA(257-264)-specific Th1 response and cytotoxic T lymphocyte (CTL) that results in significant inhibition of E.G7-OVA tumor growth. The ability of Hsp70L1 to activate DCs indicates its potential as a novel adjuvant for use with peptide immunizations; the Hsp70L1 antigen peptide hybrid may serve as a more effective vaccine for the control of cancer and infectious diseases.展开更多
Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbr...Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbred miceof different age groups and indirect immunofluorescent tech-nique. A significant decline of both Ia^+DECs and Thy-1^+DECs in the mice of the aged group was demonstrated anddifferent densities and different distribution patterns betweenIa^+DECs and Thy-1^+DECs were obserged. These findingsmay imply that the decline of both Ia^+DECs and Thy-1^+DECs in the aged group may reflect the alterations of im-mune response in aging.展开更多
Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles o...Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.Results We found that Tmod1-deficient mDCs showed deficient antigen-presenting ability and they failed to express enough MHC-Ⅱ,co-stimulated molecules(CD80/86,CD40)and CCR7 on their cell surface.The secretions of the inflammatory cytokines IL-6 and IFN-γwere decreased while the anti-inflammatory cytokines IFN-βand IL-10 were increased in the supernatant of Tmod1-deficient mDCs.As compared to DCs of wild type mice,the migration ability of DCs from Tmod1 knockout mice were dramatically damaged including their free migration and CCL19 mediated chemotaxis migration.However,we found that Tmod1 knockout had no effects on the imDCs’endocytosis ability.Furthermore,Tmod1 knockout DCs showed higher osmotic fragility,lower Young’s modulus,less F-actin content and shorter dendritic pseudopodia.Under LPS stimulation,the phosphorylation level of p65 and p38 were significantly downregulated in Tmod1 knockout mice while the expression of p-IRF3 was upregulated.Conclusions These results indicated that Tmodl knockout leads to deficient antigen-presenting ability and impaired migration of DCs as well as their biophysical properties.The underlying mechanisms are due to the inhibitions of the TLR4-mediated NF-κB and p38 MAPK singling pathway and the activation of the IRF3 signaling pathway,as well as the disturbed reorganization of the F-actin cytoskeleton.Our results provide a new insight on the functions of Tmod1 which can affect the DCs’immune functions and biophysical properties through regulating the TLR4-mediated singling pathways and cytoskeleton remodeling.展开更多
To construct a lentiviral shRNA vector targeting rat CD40 gene and detect its effectiveness of gene silencing in dendritic cells(DCs), specific siRNA targets with short hairpin frame were designed and synthesized ac...To construct a lentiviral shRNA vector targeting rat CD40 gene and detect its effectiveness of gene silencing in dendritic cells(DCs), specific siRNA targets with short hairpin frame were designed and synthesized according to the mRNA sequence of rat CD40 gene. DNA oligo was cloned into lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. The verified plasmids were transfected into 293T cells that over-express recombinant CD40 in order to select the most effective siRNA targets, shRNA lentiviruses from the selected constructs were propagated and harvested with a virus packaging system, and the virus titers were determined. Western blot and Real-time PCR were performed to determine CD40 expression level in the virusinfected dendritic cells. PCR and sequencing analyses reveal that shRNA plasmids of four targets were successfully constructed. The optimal interfering target was selected, and the virus with a titer of 5 × 10^7 TU/mL was successfully packaged. CD40 expression in rat DCs was knockdown at both mRNA and protein levels by virus infection. In comparison to that of control groups, CD40 mRNA expression and protein expression were decreased by 60.9% and 61.2%, respectively. We have successfully constructed recombinant lentiviral shRNA expression vector targeting rat CD40 gene that can effectively down-regulate CD40 gene expression at mRNA and protein levels in rat DC.展开更多
Objective: To investigate the immunotherapy efficacy of fusion cells (dendritic-C6anti-TGF-β1 cells) in the treatment of intraeranial gliomas. Methods: Dendritic cells were isolated from rat bone-marrow precursor...Objective: To investigate the immunotherapy efficacy of fusion cells (dendritic-C6anti-TGF-β1 cells) in the treatment of intraeranial gliomas. Methods: Dendritic cells were isolated from rat bone-marrow precursors stimulated in vitro with granuloeyte-macrophage colony-stimulating factor (GM-CSF) and Interleukin-4 (IL-4). C6anti-TGF-β1 cells originally from C6 cell line of a rat glioblastoma were transfected with plasmid of TGF-β1 anti-sense gene. Fusions of dendritic cells and C6anti-TGF-β1 cells were prepared by polyethylene glycol (PEG). The DC/C6anti-TGF-β1 fusion cells were observed and confirmed by fight microscopy and scanning electron microscopy. Experimental rats were divided into three groups at random: C6 cells (Ⅰ), dendritic-C6anti-TGF-β1 fusion cells and C6 cells (Ⅱ) and IMDM medium only (Ⅲ). The cells were injected into right parietal lobe region of the rat with stereotaxic technique. Histology, tumor necrosis and survival time were evaluated. Results: Compared with the rats that received C6 cells (survival median time was less than 20 days, tumor region was seen in all fields of observed), the rats injected with dendritic-C6anti-TGF-β1 fusion cells and C6 cells got a more prolonged life span (more than 59 days), as well as less tumor region (5.01%-6.2%). There was no tumor necrosis, but some glias were seen in surroundings. All rats were survived and no necrosis was observed in negative control group. Statistical analysis showed that group Ⅱ had significant difference compared with group Ⅰ. Conclusions: Dendritic-C6anti-TGF-β1 fusion cells could prolong the life span of rats, providing a strategy to achieve an antitumor response against tumors in the central nervous system.展开更多
Objective: To investigate the antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer mice model with the survival time of mice calculated and the tumor size measured in DC vaccine t...Objective: To investigate the antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer mice model with the survival time of mice calculated and the tumor size measured in DC vaccine therapy. Methods: C57BL/6 mice were immunized on the dorsal flank by s.c. inoculation of Lysate-DC, ova-DC, and non-DC on day -7. On day 0, 2× 10^6cells of RM-1 tumor cells (H-2b) were injected s.c. in C57BL/6 mice pre-treated by s.c. inoculation of modified DCs, correspondingly. DTH assay was performed with modified DCs. In partial test, for the determination of which immune cells were required for antitumor activity, mice were immunodepleted of CD4, CDS, or natural killer (NK) NK1.1 cells with the corresponding monoclonal antibodies. The survival time of nude mice loaded with tumor cells was calculated and the size of tumor measured. Results: In RM-1 mice prostate cancer model, immunized with lysate-DC, compared with ova-DC and non-DC, the pre-infection vaccine resulted in 100% clearance of primary tumors, whereas on day 0 of injection vaccine cleared 40-60% of primary tumors. On day 0, C57BL/6 mice (H-2b) were immunized with Lysate-DC, compared with ova-DC and non-DC by caudal vein injection, then on day 15, RM-1 cells were inoculated. On day 30, average diameters of tumor in different groups of modified DC were 23.7±5.4 mm, 22.1±4.9 mm, 4.3±2.6 mm, respectively. Lysate-DC, compared with ova-DC and non-DC, can greatly depressed RM-1 tumor cell growth (P〈0.01). The mean survival time of C57BL/6 mice in Lysate-DC, ova-DC and non-DC groups were 15.8±2.6, 16.6±3.2, 39.0±5.6, respectively, and there was a significant difference in the mean survival time in lysate-DC group between ova-DC and non-DC group (P〈0.01). DTH test showed that lysate-DC could prime T lymphocyte and elicit tumor antigen specific immune response, and over 80% mice in groups of lysate-DC showed obvious swelling in their foot pad. This response was strengthened with repeating inoculation, whereas DTH response was not seen in control group. In vivo depletion of NK cells resulted in a 40-60% reduction in growth suppression within the primary tumor, and depletion of CD4^+ cells resulted in a 20% reduction in growth suppression. Conclusion: The minor lysate-pulsed dendritic cells vaccine could elicit antitumor activity in RM-1 loaded C57BL/6 mice, and prolong the duration of RM-1 loaded C57BL/6 mice. So DC-based immunotherapy with hormone-refractory prostate carcinoma yielded protective immunity, generated efficient cellular antitumor responses, thereby providing further preclinical support for feasible immunotherapy approaches for prostate cancer.展开更多
Fc-epsilon receptor (CD23)-expressing follicular dendritic cells is a main prognostic factor in follicular lymphoma. Falaleeva N. A., Osmanov E. A., Tupitsyn N. N. Federal State Budgetary Institute N. N. Blokhin Russi...Fc-epsilon receptor (CD23)-expressing follicular dendritic cells is a main prognostic factor in follicular lymphoma. Falaleeva N. A., Osmanov E. A., Tupitsyn N. N. Federal State Budgetary Institute N. N. Blokhin Russian Cancer Research Center, Health Ministry of Russian Federation, Moscow, Russia SUMMARY Follicular dendritic cells, expressing FcεRII or CD23 (FcεRIIFDCs) as a component of non-tumor environment have been studied in 232 follicular lymphoma (FL) patients. FcεRIIFDCs were found in 87.5% of follicular lymphoma cases and were associated with a nodular pattern of tumor growth (p = 0.000), but not the cytological grade of lymphoma. There were no associations of FcεRIIFDC with clinical prognostic factors (FLIPI indices) or with bone marrow involvement in FL patients by histology. The presence of FcεRIIFDCs in tumor tissue was an independent prognostic factor according to treatment results, i.e. frequency of CR, duration of OS and PFS. Bone marrow involvement significantly worsened the prognosis in FcεRIIFDC-positive group of patients. We suggest a new prognostic index (FDC-IP) that allows biochemical identification of the following patient groups: FcεRIIFDC-positive patients without bone marrow involvement (good prognosis), FcεRIIF-DC-positive patients with bone marrow involvement (intermediate prognosis), FcεRIIFDC-negative patients (poor prognosis). These 3 groups significantly differ (p = 0.000) both in OS and in PFS. This is the first evidence of the possibility to assess tumor behavior and treatment results in FL according to lymphoma biochemical and other than clinical parameters.展开更多
Objective: The aim of this study was to study the expression and the clinical significance of B7-H1 on dendritic cells (DCs) in peripheral blood from patients with bladder cancer. Methods: Peripheral blood mononuc...Objective: The aim of this study was to study the expression and the clinical significance of B7-H1 on dendritic cells (DCs) in peripheral blood from patients with bladder cancer. Methods: Peripheral blood mononuclear cell were disparted from 30 bladder cancer patients and 7 healthy controls by density gradient centfifugation and then co-cultured. The expres- sion of B7-H1 on DCs were analyzed by flow cytometry. Results: Expression of BT-H1 on DCs in bladder cancer was higher than healthy controls (P 〈 0.01). And the expression were strongly associated with the pathological grade and clinical stage of bladder cancer (P 〈 0,05). Conclusion: The up-regulation of B7-H1 on DCs was strongly associated with neoplastic progres-sion of bladder cancer. B7-H1/programmed death (PD)-1 signal pathway may also play an important role in immune escape of bladder cancer during initial phase of T cell immune response.展开更多
Dermal mucinosis is often associated with collagen diseases such as rheumatoid arthritis, lupus erythematosus, and dermatomyositis, in addition to autoimmune thyroiditis. We report eight cases of dermal mucin depositi...Dermal mucinosis is often associated with collagen diseases such as rheumatoid arthritis, lupus erythematosus, and dermatomyositis, in addition to autoimmune thyroiditis. We report eight cases of dermal mucin deposition secondary to typical dermatomyositis with cutaneous lesions known as heliotrope rash and Gottron's papules. Striking mucin deposition was observed in both the papillary dermis and reticular dermis of all biopsy specimens. Immunohistochemical analysis showed that CD34+ dermal dendritic cells(DDCs) in the perilesional area in combination with vimentin+ cells within the mucinous lesion might be important in giving rise to abnormal deposition of dermal mucin. On the other hand, numbers of factor ⅩⅢa+ DDCs and tryptase+ mast cells were reduced within and surrounding the mucin deposition, as compared with those in the dermis of normal controls. A pathogenic mechanism of dermal mucin deposition is proposed.展开更多
基金supported by the National Natural Science Foundation of China(No.81972681,82103677)Tianjin Education Commission Research Plan Project(No.2021KJ201)+1 种基金Shenzhen High-level Hospital Construction Fund(No.G2022139)Tianjin Key Medical Discipline(Specialty)Construction Project(No.TJYXZDXK-009A).
文摘Objective:CD8+T cells are the key effector cells in the anti-tumor immune response.The mechanism underlying the infiltration of CD8+T cells in esophageal squamous cell carcinoma(ESCC)has not been clearly elucidated.Methods:Fresh ESCC tissues were collected and grouped according to the infiltration density of CD8+T cells.After the transcriptome sequencing on these samples and the combined analyses with The Cancer Genome Atlas(TCGA)ESCC data,a secreted protein DEFB1 was selected to explore its potential role in the infiltration of CD8+T cells.Bioinformatics analyses,histological verification and in vitro experiments were then performed.Results:DEFB1 was highly expressed in ESCC,and the high expression of DEFB1 was an independent risk factor for overall survival.Since the up-regulation or down-regulation of DEFB1 did not affect the proliferation,migration and apoptosis of ESCC cells,we speculated that the oncogenic effect of DEFB1 was achieved by regulating microenvironmental characteristics.Bioinformatics analyses suggested that DEFB1 might play a major role in the inflammatory response and anti-tumor immune response,and correlate to the infiltration of immature dendritic cell(imDC)in ESCC.Histological analyses further confirmed that there were less CD8+T cells infiltrated,less CD83+mature DC(mDC)infiltrated and more CD1a+imDC infiltrated in those ESCC samples with high expression of DEFB1.After the treatment with recombinant DEFB1 protein,the maturation of DC was hindered significantly,followed by the impairment of the killing effects of T cells in both 2D and 3D culture in vitro.Conclusions:Tumor-derived DEFB1 can inhibit the maturation of DC and weaken the function of CD8+T cells,accounting for the immune tolerance in ESCC.The role of DEFB1 in ESCC deserves further exploration.
基金Supported by Grants From the German Research Foundation DFG[UGS(FOR 876),FO,HCR]Research Grants From the University of Regensburg,Germany as part of the ReForM-program(UGS,FO)
文摘AIM:To investigate possible differences in dendritic cells(DC)within intestinal tissue of mice before and after induction of colitis. METHODS:Mucosal DC derived from intestinal tissue,as well as from mesenteric lymph nodes and spleen,were analyzed by fluorescence activated cell sorting(FACS) analysis.Supernatants of these cells were analyzed for secretion of different pro-and anti-inflammatory cytokines. Immunohistochemistry and immunofluorescence were performed on cryosections of mucosal tissue derived from animals with colitis as well as from healthy mice. RESULTS:It was shown that DC derived from healthy intestinal lamina propria(LP)represented an immature phenotype as characterized by low-level expression of costimulatory cytokines.In contrast to DC from spleen and mesenteric lymph nodes(MLN)that secreted proinflammatory cytokines,LP-DC produced high levels of the anti-inflammatory cytokine IL-10.After induction of mu-rine colitis in a CD4 + CD62L + transfer model or in chronic Dextran sulfate sodium-colitis,a marked increase of activated CD80 + DC could be observed within the inflamed colonic tissue.Interestingly,in contrast to splenic DC,a significant population of DC within MLN and colonic LP expressed the mucosal integrin CD103 which was lost during colitis. CONCLUSION:The constitutive secretion of antiinflammatory cytokines by immature DC within the intestinal LP might regulate the homeostatic balance between mucosal immunity and tolerance.CD103 + DC could mediate this important function.
基金Supported by The National Key Research and Development Programs of China,No.2022YFC2603801Maternal and Child Health Project of Jiangsu Province,No.F201717+1 种基金Doctor Project of Affiliated Hospital of Jiangsu University,No.jdfyrc2019003Clinical and Virology Study of 2019-ncov Infection in Patients with Moderate to Severe Psoriasis,No.Jdfyxgzx005.
文摘BACKGROUND Blastic plasmacytoid dendritic cell tumor(BPDCN)is a rare and highly invasive lymphohematopoietic tumor that originates from plasmacytoid dendritic cells.BPDCN has an extremely poor prognosis.Skin lesions are usually the first manifestation of BPDCN,although the tumor may also invade the bone marrow,lymph nodes,peripheral blood,and other parts of the body,leading to several other manifestations,requiring further differentiation through skin biopsy and immunohistochemistry.CASE SUMMARY In the present paper,the cases of 2 patients diagnosed with BPDCN are discussed.The immunohistochemistry analysis of these 2 patients revealed positivity for CD4,CD56,and CD123.Currently,no standard chemotherapy regimen is available for BPDCN.Therefore,intensive therapy for acute lymphoblastic leukemia was applied as the treatment method for these 2 cases.CONCLUSION Although allogeneic bone marrow transplantation could be further effective in prolonging the median survival the ultimate prognosis was unfavorable.Future treatment modalities tailored for elderly patients will help prolong survival.
文摘The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These systems can induce specific cell reactions,promote specific tissue functions,and serve as valuable tools for research in tissue engineering,regenerative medicine,and drug discovery.This paper discusses current developments in the field of three-dimensional cell culture and the potential applications of 3D type 1 collagen gels to enhance the growth and maturation of dendritic cells.
基金This project was supported by a grant from National Natural Sciences Foundation of China (No. 30271300)
文摘Objective: To observe the lethal effect of T cells activated by dendritic cells (DCs) with MUC1 gene transfection (MUC1-DCs) on BIU-87 cells in vitro, and to evaluate the possibility of dendritic cells transfected by MUC1 gene as a vaccine for bladder cancer immunotherapy. Methods: MUC1 was successfully transfected into cultured human blood-derived dendritic cell with lipofectin. The transfection efficiency and immunocompetence were detected by flow cytometry and MTT colourmetry. T lymphocytes activated by MUC1-DCs were used to kill BIU-87 cell lines and normal bladder epithelium in vitro and the killing rate was evaluated by MTT colourmetry. Results: There was significant lethal effect on the BIU-87 cells and normal bladder epithelium with T cells activated by MUCl-transfected DCs, but the lethal effect on the BIU-87 cells significantly exceeded that on normal bladder epithelium (P〈0.05). The lethal effect on BIU-87 cells with T cells activated by MUCl-transfected DCs significantly exceeded that with T cells activated by no-transfection DCs (P〈0.01). Conclusion: T cells activated by MUC1-DCs could induce killing action to BIU-87 cell lines. MUC1 gene could be used as a target for bladder cancer immunotherapy.
基金Supported by a grant from the Natural Science Foundation of Jiangsu Province,China,No.BK2008237
文摘AIM:To investigate the function of PU.1-silenced semimature dendritic cells(DCs)and the possibility of utilizing cell immunity in rat intestinal transplantation.METHODS:DCs were isolated from the bone marrow of F344 rats and cultured using the adherent method.The PU.1 gene was knocked down in DCs using small interfering RNAs(siRNAs)for 24 h,and the cells were then incubated with lipopolysaccharide for 48 h.The PU.1 siRNA that had the highest silencing efficiency was screened using reverse transcription-polymerase chain reaction and Western blot for further study.The tolerance capacity was analyzed and compared between PU.1-silenced DCs(siRNA PU.1 group),negative control-silenced DCs(siRNA NC group)and immature DCs(control group)both in vitro and in vivo.CONCLUSION:Blocking expression of the PU.1 gene in vitro led to a reduction in DC maturation and an increased tolerance capability.PU.1-silenced DCs expressed moderate levels of major histocompatibility complex(MHC)-Ⅱand low levels of co-stimulatory molecules,and produced more interleukin(IL)-10,but less IL-12.Compared with the negative control group,surface molecules cluster of differentiation 80(CD80),CD86 and MHC-Ⅱin the siRNA PU.1 group were 27.0%±5.6%,23.6%±4.8%and 36.8%±6.8%,respectively,and showed a significantly lower trend(P<0.05).In vivo treatment of recipients with PU.1-silenced DCs injected before intestinal transplantation(siRNA PU.1group),significantly prolonged allograft survival and resulted in better tissue histopathology compared with the siRNA NC group and control group.Mean survival time after transplantation was 14.3±3.3 d in the siRNA PU.1 group(P<0.05).CONCLUSION:PU.1-silenced semi-mature DCs induced partial immune tolerance both in vitro and in vivo,which could be used as a new strategy to promote transplantation tolerance.
基金A basic science grant from CONACyT,No.285480the Department of Clinical Immunology Research of the Biochemical Sciences Faculty.
文摘Type 1 diabetes(T1D)is an autoimmune disease characterized by the destruction of insulin-producingβ-cells of the pancreatic islets by autoreactive T cells,leading to high blood glucose levels and severe long-term complications.The typical treatment indicated in T1D is exogenous insulin administration,which controls glucose levels;however,it does not stop the autoimmune process.Various strategies have been implemented aimed at stoppingβ-cell destruction,such as cellular therapy.Dendritic cells(DCs)as an alternative in cellular therapy have gained great interest for autoimmune disease therapy due to their plasticity to acquire immunoregulatory properties both in vivo and in vitro,performing functions such as anti-inflammatory cytokine secretion and suppression of autoreactive lymphocytes,which are dependent of their tolerogenic phenotype,displayed by features such as semimature phenotype,low surface expression of stimulatory molecules to prime T cells,as well as the elevated expression of inhibitory markers.DCs may be obtained and propagated easily in optimal amounts from peripheral blood or bone marrow precursors,such as monocytes or hematopoietic stem cells,respectively;therefore,various protocols have been established for tolerogenic(tol)DCs manufacturing for therapeutic research in the treatment of T1D.In this review,we address the current advances in the use of tolDCs for T1D therapy,encompassing protocols for their manufacturing,the data obtained from preclinical studies carried out,and the status of clinical research evaluating the safety,feasibility,and effectiveness of tolDCs.
基金Project supported by the National Natural Science Foundation of China (No. 30270571)Doctor Degree Point Foundation of Ministry of Education, China (No. 20020335080)
文摘Objective: To explore the effects of transforming growth factor (51 (TGF-β1) on dendritic cells (DC). Methods: Murine bone marrow cells were cultured with GM-CSF and TGF-β1 to develop TGF-β1-treated DC (TGFβ-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 and IL-12p70 protein was detected by ELISA. The expression of Toll-like receptor 4 (TLR4) was analyzed by semi quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and FCM. Results: Compared to immature DC (imDC) cultured by GM-CSF alone, the TGFβ-DC express lower CD80, CD86,I-Ab and CD40. The TGFβ-DC were resistant to maturation with LPS. Maturation resistance was evident from a failure to up-regulate co-stimulatory molecules (CMs), to stimulate larger T cells proliferation and to enhance secretion of IL-12p70. We also found that TGF-β1 could down-regulate TLR4 expression on TGFβ-DC. Conclusion: TGFβ-DC are resistant to maturation stimulus (LPS) and might have some correlation with the down-modulation of TLR4 expression.
基金Supported by the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education,Science and Technology,South Korea,No.NRF-2018R1D1A1B07043350
文摘BACKGROUND Enterotoxigenic Bacteroides fragilis(ETBF)causes colitis and diarrhea,and is considered a candidate pathogen in inflammatory bowel diseases as well as colorectal cancers.These diseases are dependent on ETBF-secreted toxin(BFT).Dendritic cells(DCs)play an important role in directing the nature of adaptive immune responses to bacterial infection and heme oxygenase-1(HO-1)is involved in the regulation of DC function.AIM To investigate the role of BFT in HO-1 expression in DCs.METHODS Murine DCs were generated from specific pathogen-free C57BL/6 and Nrf2−/−knockout mice.DCs were exposed to BFT,after which HO-1 expression and the related signaling factor activation were measured by quantitative RT-PCR,EMSA,fluorescent microscopy,immunoblot,and ELISA.RESULTS HO-1 expression was upregulated in DCs stimulated with BFT.Although BFT activated transcription factors such as NF-κB,AP-1,and Nrf2,activation of NF-κB and AP-1 was not involved in the induction of HO-1 expression in BFT-exposed DCs.Instead,upregulation of HO-1 expression was dependent on Nrf2 activation in DCs.Moreover,HO-1 expression via Nrf2 in DCs was regulated by mitogenactivated protein kinases such as ERK and p38.Furthermore,BFT enhanced the production of reactive oxygen species(ROS)and inhibition of ROS production resulted in a significant decrease of phospho-ERK,phospho-p38,Nrf2,and HO-1 CONCLUSION These results suggest that signaling pathways involving ROS-mediated ERK and p38 mitogen-activated protein kinases-Nrf2 activation in DCs are required for HO-1 induction during exposure to ETBF-produced BFT.
文摘Heat shock proteins (HSPs) are reported to act as effective adjuvants to elicit anti-tumor and anti-infection immunity. Here, we report that Hsp70-like protein 1 (Hsp70L1), a novel HSP derived from human dendritic cells (DCs), has potent adjuvant effects that polarize responses toward Th1. With a calculated molecular weight of 54.8 kDa, Hsp70L1 is smaller in size than Hsp70 but resembles it both structurally and functionally. Hsp70L1 shares common receptors on DCs with Hsp70 and can interact with DCs, promoting DC maturation and stimulating secretion of the proinflammatory cytokines interleukin 12p70 (IL-12p70), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), and the chemokines IP-10, macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, and normal T cell expressed and secreted (RANTES). The induction of interferon-gamma-inducible protein 10 (IP-10) secretion by Hsp70L1 is not shared by Hsp70, and other functional differences include more potent stimulation of DC IL-12p70, CC-chemokine, and CCR7 and CXCR4 expression by Hsp70L1. Immunization of mice with the hybrid peptide Hsp70L1-ovalbumin(OVA)(257-264) induces an OVA(257-264)-specific Th1 response and cytotoxic T lymphocyte (CTL) that results in significant inhibition of E.G7-OVA tumor growth. The ability of Hsp70L1 to activate DCs indicates its potential as a novel adjuvant for use with peptide immunizations; the Hsp70L1 antigen peptide hybrid may serve as a more effective vaccine for the control of cancer and infectious diseases.
文摘Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbred miceof different age groups and indirect immunofluorescent tech-nique. A significant decline of both Ia^+DECs and Thy-1^+DECs in the mice of the aged group was demonstrated anddifferent densities and different distribution patterns betweenIa^+DECs and Thy-1^+DECs were obserged. These findingsmay imply that the decline of both Ia^+DECs and Thy-1^+DECs in the aged group may reflect the alterations of im-mune response in aging.
基金funded by the National Natural Science Foundation of China ( 31660258,31771014, 31860262,31570938,31260227)the Science and Technology Foundation of Guizhou Province ( 2019-2787,2018-1412, 2016-5676,2017-5718)+2 种基金the Science and Technology Innovative Talent Team of Guizhou Province ( 2015-4021)the 2011 Collaborative Innovation Program of Guizhou Province ( 2015-04 )the Cell and Gene Engineering Innovative Research Groups of Guizhou Province ( KY-2016-031)
文摘Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.Results We found that Tmod1-deficient mDCs showed deficient antigen-presenting ability and they failed to express enough MHC-Ⅱ,co-stimulated molecules(CD80/86,CD40)and CCR7 on their cell surface.The secretions of the inflammatory cytokines IL-6 and IFN-γwere decreased while the anti-inflammatory cytokines IFN-βand IL-10 were increased in the supernatant of Tmod1-deficient mDCs.As compared to DCs of wild type mice,the migration ability of DCs from Tmod1 knockout mice were dramatically damaged including their free migration and CCL19 mediated chemotaxis migration.However,we found that Tmod1 knockout had no effects on the imDCs’endocytosis ability.Furthermore,Tmod1 knockout DCs showed higher osmotic fragility,lower Young’s modulus,less F-actin content and shorter dendritic pseudopodia.Under LPS stimulation,the phosphorylation level of p65 and p38 were significantly downregulated in Tmod1 knockout mice while the expression of p-IRF3 was upregulated.Conclusions These results indicated that Tmodl knockout leads to deficient antigen-presenting ability and impaired migration of DCs as well as their biophysical properties.The underlying mechanisms are due to the inhibitions of the TLR4-mediated NF-κB and p38 MAPK singling pathway and the activation of the IRF3 signaling pathway,as well as the disturbed reorganization of the F-actin cytoskeleton.Our results provide a new insight on the functions of Tmod1 which can affect the DCs’immune functions and biophysical properties through regulating the TLR4-mediated singling pathways and cytoskeleton remodeling.
基金Supported by the National Natural Science Foundation of China(No.30670301)Science and Technology Services of Jilin Province, China(No.20050408-1)+1 种基金PhD Scientific Research Foundation of Ministry of Education of China(No.20050183069)the Jilin Province Talent Development Foundation(No.JRJB2007-2)
文摘To construct a lentiviral shRNA vector targeting rat CD40 gene and detect its effectiveness of gene silencing in dendritic cells(DCs), specific siRNA targets with short hairpin frame were designed and synthesized according to the mRNA sequence of rat CD40 gene. DNA oligo was cloned into lentiviral expression vector, and then PCR and sequencing analyses were conducted to verify the constructs. The verified plasmids were transfected into 293T cells that over-express recombinant CD40 in order to select the most effective siRNA targets, shRNA lentiviruses from the selected constructs were propagated and harvested with a virus packaging system, and the virus titers were determined. Western blot and Real-time PCR were performed to determine CD40 expression level in the virusinfected dendritic cells. PCR and sequencing analyses reveal that shRNA plasmids of four targets were successfully constructed. The optimal interfering target was selected, and the virus with a titer of 5 × 10^7 TU/mL was successfully packaged. CD40 expression in rat DCs was knockdown at both mRNA and protein levels by virus infection. In comparison to that of control groups, CD40 mRNA expression and protein expression were decreased by 60.9% and 61.2%, respectively. We have successfully constructed recombinant lentiviral shRNA expression vector targeting rat CD40 gene that can effectively down-regulate CD40 gene expression at mRNA and protein levels in rat DC.
文摘Objective: To investigate the immunotherapy efficacy of fusion cells (dendritic-C6anti-TGF-β1 cells) in the treatment of intraeranial gliomas. Methods: Dendritic cells were isolated from rat bone-marrow precursors stimulated in vitro with granuloeyte-macrophage colony-stimulating factor (GM-CSF) and Interleukin-4 (IL-4). C6anti-TGF-β1 cells originally from C6 cell line of a rat glioblastoma were transfected with plasmid of TGF-β1 anti-sense gene. Fusions of dendritic cells and C6anti-TGF-β1 cells were prepared by polyethylene glycol (PEG). The DC/C6anti-TGF-β1 fusion cells were observed and confirmed by fight microscopy and scanning electron microscopy. Experimental rats were divided into three groups at random: C6 cells (Ⅰ), dendritic-C6anti-TGF-β1 fusion cells and C6 cells (Ⅱ) and IMDM medium only (Ⅲ). The cells were injected into right parietal lobe region of the rat with stereotaxic technique. Histology, tumor necrosis and survival time were evaluated. Results: Compared with the rats that received C6 cells (survival median time was less than 20 days, tumor region was seen in all fields of observed), the rats injected with dendritic-C6anti-TGF-β1 fusion cells and C6 cells got a more prolonged life span (more than 59 days), as well as less tumor region (5.01%-6.2%). There was no tumor necrosis, but some glias were seen in surroundings. All rats were survived and no necrosis was observed in negative control group. Statistical analysis showed that group Ⅱ had significant difference compared with group Ⅰ. Conclusions: Dendritic-C6anti-TGF-β1 fusion cells could prolong the life span of rats, providing a strategy to achieve an antitumor response against tumors in the central nervous system.
基金Supported by medical funds of Shanghai Science and Technology Commission
文摘Objective: To investigate the antitumor activity of tumor lysate-pulsed dendritic cells vaccine in RM-1 prostate cancer mice model with the survival time of mice calculated and the tumor size measured in DC vaccine therapy. Methods: C57BL/6 mice were immunized on the dorsal flank by s.c. inoculation of Lysate-DC, ova-DC, and non-DC on day -7. On day 0, 2× 10^6cells of RM-1 tumor cells (H-2b) were injected s.c. in C57BL/6 mice pre-treated by s.c. inoculation of modified DCs, correspondingly. DTH assay was performed with modified DCs. In partial test, for the determination of which immune cells were required for antitumor activity, mice were immunodepleted of CD4, CDS, or natural killer (NK) NK1.1 cells with the corresponding monoclonal antibodies. The survival time of nude mice loaded with tumor cells was calculated and the size of tumor measured. Results: In RM-1 mice prostate cancer model, immunized with lysate-DC, compared with ova-DC and non-DC, the pre-infection vaccine resulted in 100% clearance of primary tumors, whereas on day 0 of injection vaccine cleared 40-60% of primary tumors. On day 0, C57BL/6 mice (H-2b) were immunized with Lysate-DC, compared with ova-DC and non-DC by caudal vein injection, then on day 15, RM-1 cells were inoculated. On day 30, average diameters of tumor in different groups of modified DC were 23.7±5.4 mm, 22.1±4.9 mm, 4.3±2.6 mm, respectively. Lysate-DC, compared with ova-DC and non-DC, can greatly depressed RM-1 tumor cell growth (P〈0.01). The mean survival time of C57BL/6 mice in Lysate-DC, ova-DC and non-DC groups were 15.8±2.6, 16.6±3.2, 39.0±5.6, respectively, and there was a significant difference in the mean survival time in lysate-DC group between ova-DC and non-DC group (P〈0.01). DTH test showed that lysate-DC could prime T lymphocyte and elicit tumor antigen specific immune response, and over 80% mice in groups of lysate-DC showed obvious swelling in their foot pad. This response was strengthened with repeating inoculation, whereas DTH response was not seen in control group. In vivo depletion of NK cells resulted in a 40-60% reduction in growth suppression within the primary tumor, and depletion of CD4^+ cells resulted in a 20% reduction in growth suppression. Conclusion: The minor lysate-pulsed dendritic cells vaccine could elicit antitumor activity in RM-1 loaded C57BL/6 mice, and prolong the duration of RM-1 loaded C57BL/6 mice. So DC-based immunotherapy with hormone-refractory prostate carcinoma yielded protective immunity, generated efficient cellular antitumor responses, thereby providing further preclinical support for feasible immunotherapy approaches for prostate cancer.
文摘Fc-epsilon receptor (CD23)-expressing follicular dendritic cells is a main prognostic factor in follicular lymphoma. Falaleeva N. A., Osmanov E. A., Tupitsyn N. N. Federal State Budgetary Institute N. N. Blokhin Russian Cancer Research Center, Health Ministry of Russian Federation, Moscow, Russia SUMMARY Follicular dendritic cells, expressing FcεRII or CD23 (FcεRIIFDCs) as a component of non-tumor environment have been studied in 232 follicular lymphoma (FL) patients. FcεRIIFDCs were found in 87.5% of follicular lymphoma cases and were associated with a nodular pattern of tumor growth (p = 0.000), but not the cytological grade of lymphoma. There were no associations of FcεRIIFDC with clinical prognostic factors (FLIPI indices) or with bone marrow involvement in FL patients by histology. The presence of FcεRIIFDCs in tumor tissue was an independent prognostic factor according to treatment results, i.e. frequency of CR, duration of OS and PFS. Bone marrow involvement significantly worsened the prognosis in FcεRIIFDC-positive group of patients. We suggest a new prognostic index (FDC-IP) that allows biochemical identification of the following patient groups: FcεRIIFDC-positive patients without bone marrow involvement (good prognosis), FcεRIIF-DC-positive patients with bone marrow involvement (intermediate prognosis), FcεRIIFDC-negative patients (poor prognosis). These 3 groups significantly differ (p = 0.000) both in OS and in PFS. This is the first evidence of the possibility to assess tumor behavior and treatment results in FL according to lymphoma biochemical and other than clinical parameters.
文摘Objective: The aim of this study was to study the expression and the clinical significance of B7-H1 on dendritic cells (DCs) in peripheral blood from patients with bladder cancer. Methods: Peripheral blood mononuclear cell were disparted from 30 bladder cancer patients and 7 healthy controls by density gradient centfifugation and then co-cultured. The expres- sion of B7-H1 on DCs were analyzed by flow cytometry. Results: Expression of BT-H1 on DCs in bladder cancer was higher than healthy controls (P 〈 0.01). And the expression were strongly associated with the pathological grade and clinical stage of bladder cancer (P 〈 0,05). Conclusion: The up-regulation of B7-H1 on DCs was strongly associated with neoplastic progres-sion of bladder cancer. B7-H1/programmed death (PD)-1 signal pathway may also play an important role in immune escape of bladder cancer during initial phase of T cell immune response.
基金A research grant from the Japan Society for the Promotion of Science(25461695 to Masahiko Muto)a grant for Research on Measures for Intractable Diseases(to Masahiko Muto)from the Ministry of Health,Labour and Welfare,Japan
文摘Dermal mucinosis is often associated with collagen diseases such as rheumatoid arthritis, lupus erythematosus, and dermatomyositis, in addition to autoimmune thyroiditis. We report eight cases of dermal mucin deposition secondary to typical dermatomyositis with cutaneous lesions known as heliotrope rash and Gottron's papules. Striking mucin deposition was observed in both the papillary dermis and reticular dermis of all biopsy specimens. Immunohistochemical analysis showed that CD34+ dermal dendritic cells(DDCs) in the perilesional area in combination with vimentin+ cells within the mucinous lesion might be important in giving rise to abnormal deposition of dermal mucin. On the other hand, numbers of factor ⅩⅢa+ DDCs and tryptase+ mast cells were reduced within and surrounding the mucin deposition, as compared with those in the dermis of normal controls. A pathogenic mechanism of dermal mucin deposition is proposed.