Objective Respiratory syncytial virus ( RSV) is a primary cause of lower respiratory tract infections in children, and is also the cause for the development of asthma primarily in infants. However,the immunological mechanisms by which RSV enhances allergic sensitization and asthma remain unclear. The aimof this study was to examine the influence of RSV-infected airway epithelial cells on the activation and functions of rat myeloid dendritic cells ( mDCs) . Methods Rat airway epithelial cells ( RAECs) were infected by RSV. Then RSV-infected RAECs were co-cultured with rat mDCs, and the expression of cytokine and maturation markers on mDCs were examined by real time PCR and flow cytometry. To confirm this functional mDC maturation, allergenic mixed lymphocyte reaction ( MLR) were performed. Results Wefound that functional maturation of mDCs was induced by RSV-treated RAECs, as shown by their enhanced levels of OX40L and thymus- and activation-regulated chemokine ( TARC) mRNAs, which increased the expressions of major histocompatibility complex II ( MHCII) and CD86 costimulatorymolecules and promotedT-cell proliferation in mixed lymphocyte reactions. Conclusion Our results suggest that RSV-infected epithelial cells promote the maturation of mDCs that might support Th2 cell polarization and contribute to the pathogenesis of asthma.
ObjectiveTo explore the antitumor effect of tumor vaccine fused from dendritic cells (DC) and Walker-256 cancer cells on implanted liver cancer in rats and the related mechanism of inhibition for tumor angiogenesis. MethodsWalker-256 cancer cells and mature DC were fused by 50% polyethylene glycol method for preparation of DC-Walker-256 fusion vaccines. Implanted liver cancer models were established through operations on healthy male SD rats at the age of 6－8 weeks. All the rats were divided into four groups, and rats in each group were injected subcutanely with fusion vaccine (group), mixed cultured cells (group), simple DC (group), and PBS (blank control group), respectively. On 28 d after making model, the rats were put to death, the tumor was observed and pathological essays were prepared. All rats’ spleens were collected and prepared into lymphocyte to detect antigenic specificity cytotoxic T lymphocyte (CTL) by enzymelinked immunosorbent spot (ELISPOT) method. The expressions of VEGF, ANG-1, ANG-2, and MVD were detected by immunohistochemistry. ResultsThe numbers of rats survived in the fusion vaccine group, mixed culture cells group, simple DC group, and blank control group was 8, 5, 6, and 3, respectively. The rats in the other three groups except for fusion vaccine group were manifested as inaction, anorexia, and gloomy fur in some degree as well as ascites. The tumorigenesis was found in all survival rats except for two in the fusion vaccine group. The weight of liver tumors of rats in the fusion vaccine group 〔(32.4±9.2) g〕 was significantly lighter than that in the mixed culture cells group 〔(67.3±5.1) g, P=0.031〕, simple DC group 〔(75.0±8.3) g, P=0.019〕, and blank control group 〔(86.6±10.5) g, P=0.008〕, respectively. The number of tumorspecific CTL of rats in the fusion vaccine group was also significantly higher than that in the other three groups (P=0.019, P=0.025, and P=0.001, respectively). The MVD of tumor tissue in the fusion vaccine group was (24.12±2.32) vessels/HP, which was significantly lower than that in the mixed culture cells group 〔(40.34±1.29) vessels/HP, P=0.025〕, simple DC group 〔(42.36±3.16) vessels/HP, P=0.035〕, and blank control group 〔(56.48±5.16) vessels/HP, P=0.006〕, respectively. The MVD of tumor tissue in the mixed cultured cells group and simple DC group was similar (P=0.165), however, which was significantly lower than that in the blank control group (P=0.040 and P=0.043). The positive rate of VEGFA protein expression was 23.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (42.5%, P=0.031), simple DC group (61.3%, P=0.019), and blank control group (89.6%, P=0.003), respectively. The positive rate of VEGF-A protein expression in the mixed cultured cells and simple DC groups was similar (P=0.089), however, which was significantly lower than that in the blank control group (P=0.027 and P=0.038). The positive rate of ANG-1 protein expression in the fusion vaccine group (43.2%) was not different from that in the mixed culture cells group (46.3%, P=0.292), simple DC group (51.3%, P=0.183), or blank control group (49.6%, P=0.179), respectively, and the difference of pairwise comparison in latter three groups was not significant (P=0.242, P=0.347, and P=0.182). The positive rate of ANG2 protein expression was 19.2% in the fusion vaccine group, which was significantly lower than that in the mixed culture cells group (62.3%, P=0.007), simple DC group (67.3%, P=0.005), and blank control group (71.6%, P=0.004), respectively, however, the difference of pairwise comparison in latter three groups was not significant (P=0.634, P=0.483, and P=0.379). ConclusionFused vaccine can induce CD8+ CTL aiming at tumor cells and establish the effective antitumor immunity in vivo and also downregulate the level of VEGF and ANG-2 to suppress tumor angiogenesis and thereby achieve the purpose of curing tumor.
Objective To study the advances in the relationship between the number of infiltrating dendritic cells and the postoperative prognosis of digestive malignant tumor. MethodsThe literature in recent years on the relationship between the number of infiltrating dendritic cells and the postoperative prognosis of digestive malignant tumor was reviewed.ResultsThe number of infiltrating dendritic cells among esophageal cancer,and gastric carcinoma,colonic cancer and pancreatic cancer was associated with a better prognosis.Conclusion The population density of dendritic cells among the malignant tissue could be regarded as an independent indicator in estimating the postoperative prognosis of malignant tumor.
Objective To study the method of obtaining a large number of dendritic cells （DC）. To study the specific cytotoxicity T lymphocyte (CTL) effect against tumor cells initiated by DC pulsed with peptide of cancer cell. Methods Development of cells with cytologic features of DC in bone marrow cultures supplemented with granulocyte macrophage-colony stimulus factor (GM-CSF) and IL-4. Determining the DC phenotype and the specific structure by electronic microscopy. The CTL effect against pancreatic carcinoma leading by the DC pulsed with tumor cells lysate in vitro was observed. Results A large number of typical DC was proliferated by supplementing with GM-CSF and IL-4 cytokines. DC had specific cell appearance and structure, and highly expressed various cell surface molecules. TNF-α had the ability of stimulating DC mature, the mature DC had the enhancing abilities of antigen presenting and IL-12 self-secreting, as well as, expressed higher levels of CD54， MHC-Ⅱ and CD86 molecules than control group (P＜0.05). T lymphoid cell stimulated by DC without tumor antigen could not recognize and kill the target cells, only if DC pulsed with peptide of cancer cell can lead a b immune response to special tumor cells. The inhibiting ratio of CTL was significantly higher than that in other groups (P＜0.01). Conclusion Bone marrow DC has b ability of inducing special CTL against determined cancer cells after they are pulsed with tumor cell lysate. DC vaccine is probably a new immunotherapeutic method against tumor in the near future.
Tumor-derived exosomes play a role in helping tumor cells with escape from immune surveillance, and it may also activate tumor-specific immune responses to eradicate tumor cells. Tumor cells release exosomes with major histocompatibility complex molecules and antigenic peptides on the surface membranes, which can induce dendritic cells (DC) and cytokine-induced killer (CIK) cells in vitro to produce the tumor antigen-specific T cells, and the obtained DC-CIK cells have a dual antitumor function with specificity and non specificity. This provides a new method for the treatment of cancers. This review briefly summarized the latest progress of adoptive immunotherapy with exosomes and DC-CIK.
【Abstract】Objective To explore the effect against gastric cancer induced by Newcastle disease virus modified autologous tumor vaccine （NDV-ATV）pulsed dendritic cells(DCs). Methods The Newcastle disease virus infected the gastric cancer lines (MNK45) and was lost its activity. Peripheral blood mononuclear cell (PBMC) were cultured under condition of recombinant human granulocyte macrophage-colony stimulating factor (1 000 u/ml)+IL-4(1 000 u/ml) + TNF-α(100 ng/ml). The tumor antigen specific cytotoxic T lymphocytes (CTL) was generated from activated autologous T cell by the Newcastle disease virus infected the MNK45 pulsed DC. And Cyto Tox 96TM in vitro assayed the cytotoxicity of CTL to MNK45. Thawed gastric cancer cell antigen were used as control in these experiments. Results The killing rate of MNK45 by antigen specific CTL reached (90.15±9.82)%, which was nearly twice as high as that of control(60.57±5.74)%. The CTL had much higher cytotoxicity to different differentiated type of gastric cancer cells such as MGC803〔(52.23±6.45)% 〕 and SGC7901〔 (61.75±8.84)%〕, as compared with LOVO〔(9.11±3.42)%〕 and HepG2 〔 (8.30±3.12)%〕tumor cells(P＜0.05). Conclusion Efficient and specific of against gastric cancer immunoreaction can be induced in virtue of NDV-ATV pulsed DCs, NDV-ATV loaded DCs might provide a new kind of theraputic means for gastric cancer.
Objective To investigate the effects of FasL gene-modified dendritic cell (DC) on the airway inflammation in mice sensitized/challenged by house dust mite (HDM) allergen.Methods FasL gene-modified DC (FasL-DC) and control DC (nontransfection DC) were administrated into HDM sensitized and challenged mice by intratracheal injection respectively，then HDM sensitized and challenged mice were sacreificed two days later.Total and differentiation cell counts and levels of interleukin-4（IL-4），IL-5 and interferon-γ(IFN-?) in bronchoalveolar lavage fluid (BALF) were detected and lung histological features were observed.Results After administration of FasL-DC，lung allergic inflammation was ameliorated while total cell counts，the percentage of eosinophil ,the levels of IL-4 and IL-5 in BALF decreased and the level of IFN-? in BALF increased.Conclusion Administrating FasL-DC into HDM sensitized/challenged mice can inhibit Th2 cells activation and ameliorate airway allergic inflammation.
Objective To explore the effect of dendritic cells (DCs) allergized by K-ras mutant peptide on expressions of chemokines CCL19, CCL22, and cytoskeletal protein fascin-1. Methods DCs were derived from peripheral blood in the presence of granuloceyte/macrophage-colony stimulating factor, interleukin (IL) -4 in vitro. The DCs were collected on day 7 after culture, and were divided into non-K-ras mutant peptide group (addition of RPMI 1604 culture solution 50 μg/ml) and K-ras mutant peptide group (addition of K-ras mutant peptide 50 μg/ml). Phenotype was identified by flow cytometry. The morphological structure was observed by scanning and transmission electron microscopies, respectively. The expressions of IL-12, CCL19, and CCL22 were tested continuously by enzyme-linked immunosorbent assay (ELISA). The expression of cytoskeletal protein fascin-1 was determined by Western blot. Results ①The expressions of CD1a, CD80, and CD86 after loading K-ras mutant peptide were higher than that before loading K-ras mutant peptide (Plt;0.01). ②The DCs with petal-like and branch-like profections after loading were observed under scanning electron microscopy; The DCs with irregular shapes, branch-like or burr-like were showed under transmission electron microscopy. ③The expressions of IL-12, CCL19, and CCL22 in the Kras mutant peptide group were higher than those in the non-K-ras mutant peptide group at different times (6, 12, 24, and 48 h) after loading Kras mutant peptide (Plt;0.01). ④The expression of fascin-1 in the K-ras mutant peptide group was also higher than that in the non-K-ras mutant peptide group (Plt;0.01). Conclusion K-ras mutant peptide can promote DC to mature and improve the expression of chemokines and cytoskeletal protein which will strengthen DC migration.
Objective To investigate the feasibility of dendritic cells ( DCs ) as vector of immunotherapy through intratracheal injection. Methods The DCs obtained from the bone marrow of BALB/ c mice were cultured and isolated with CD11c-positive magnetic beads. Then DCs were overloaded with ovalbumin peptide 323-339 ( OVA 323-339) for 24 hours. The mice in the DC-OVA group were intratrachelly injected DCs overloaded with OVA 323-339 in dose of 2 ×106 cells per mouse. The mice in thenegative control group were intratracheally injected with DCs untreated by OVA 323-339. On the second day,all mice were challenged with 1% OVA in PBS lasting for five days. The asthma animal model established by classic method was used for the positive control. Pathologic changes in lung and cell numbers in bronchoalveolar lavage fluid ( BALF) were assayed 24 hours after challenged. Results Just like the lung tissues from the mice asthma models, the lung tissues from the mice instilled with DCs overloaded with allergen OVA 323-339 showed extensive inflammatory cells infiltration, most of which were eosinophils, neutrophils and lymphocytes. The lung tissues in the DC group showed no obvious inflammation. There were more cells in BALF in the DC-OVA group than that in the DC group. OVA-specific IgE in serum from the DC-OVA group was not significantly different from that in the mice asthma models [ ( 48. 22 ±4. 76) U/mL vs. ( 52. 75 ±4. 03) U/mL, P gt;0. 05] . Conclusion DCs overloaded antigen has the ability of transferring of antigen effectively and may be used as vectors of immunotherapy.
Objective To investigate the effects of nuclear factor kappa B decoy oligodeoxynucleotides ( NF-κB decoy ODN) transfection on biological characteristics of mature dendritic cells ( mDCs) in mice. Methods Immature DCs were harvested from Balb / c mice bone marrow, followed by the incubation with antigen OVA and LPS, and mature DCs were evaluated by the expressions of CD11c and MHC-Ⅱ detected by FACS. Mature DCs were transfected with NF-κB decoy ODN and the changes of NF-κB activity after the transfection were detected by EMSA. The expressions of the costimulatory molecules( CD40,CD80 and CD86) on DCs were detected by FACS and the proliferation of T cells was tested by mixed lymphocyte reaction( MLR) . Results The mature DCs were cultured successfully. The NF-κB activity of NF-κB decoy ODN transfected DCs was decreased significantly( P lt; 0. 05) . There was no difference in the expressions of CD40 and CD80, but the expression of CD86 was decreased significantly in NF-κB decoy ODN transfection group( P lt; 0. 05) . MLR test showed that the proliferation of T lymphocyte cells was inhibited by NF-κB decoy ODN transfected DCs, but was stimulated bly by the DCs of other groups. Conclusions Mature DCs transfected with NF-κB decoy ODN could inhibit the proliferation and activation of antigenspecical T cells, which was probably related to the down-regulation of CD86 on DCs. This modified DCs might be a promising vaccine for the treatment of asthma in the future.