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ABSTRACT: In this study, we hypothesized that the mice immunized with the glycosylphosphatidylinositol (GPI) anchored 6 kDa early-secreted antigenic target (ESAT-6) DNA vaccine (ESAT-6-gpi) and the tumor vaccine B16F10-ESAT-6- gpi/IL-21 might significantly enhance immune responses and antimelanoma efficacy. Our experimental results indicated that the anti-ESAT-6 antibody induced by the DNA vaccine ESAT-6-gpi bound ESAT-6 to the surface of tumor vaccine to activate a complement classical pathway and resulted in the B16F10 tumor cell lysis and apoptosis, which served as a potential potent trigger for breaking melanomatous immune tolerance to elicit an initiation of natural anti- melanoma immunity. Our innovative approach of using the DNA vaccine ESAT-6-gpi priming and the tumor vaccine B16F10-ESAT-6-gpi/IL-21 boosting induced strong antimelanoma immunity that inhibited melanomatous growth. These findings highlighted the DNA vaccine ESAT-6-gpi as an immune-enhancer to augment the immune efficacy of the tumor vaccine B16F10-ESAT-6-gpi/IL-21 against melanoma in a mouse model. This article is protected by copyright. All rights reserved.
Scandinavian Journal of Immunology 05/2013; · 2.23 Impact Factor
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ABSTRACT: Aim: To investigate the effects of anti-ABCG2 monoclonal antibodies (mAbs) in combination with paclitaxel iron oxide nanoparticles (PTX-NPs) on CD138(-)CD34(-) multiple myeloma (MM) cancer stem cells (CSCs) in JJN3 cells. Materials & methods: PTX-NPs were prepared using the hydrophobic interaction of the polyoxypropylene chain and oleic acid on the surface of iron oxide NPs and were targeted to the ABCG2 transporter overexpressing MM CSCs with mAbs. Results: The data showed that MM CSCs have strong drug resistance and tumorigenicity compared with non-MM CSCs. PTX-NPs combined with mAbs led to a significant reduction in the tumor volume, a visible alleviation of lytic bone lesions and to a markedly increased survival rate in contrast to using a single agent in MM CSCs when it was transplanted to nonobese diabetic/severe combined immunodeficiency mice. Conclusion: This study is the first to report on the anti-MM CSC activity by PTX-NPs as a single agent or used together with anti-ABCG2 mAbs to treat MM. These findings provide a rationale for future clinical trials. Original submitted 18 June 2012; Revised submitted 29 November 2012.
Nanomedicine 03/2013; · 5.05 Impact Factor
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ABSTRACT: The goal of this study was to investigate whether glycosylphosphatidylinositol (GPI)-anchored 6 kDa early secreted antigenic target (ESAT-6) and IL-21-producing B16F10/ESAT-6-GPI-IL-21 viable vaccine would induce antitumor efficacy. Mice were immunized with B16F10/ESAT-6-GPI-IL-21 vaccine and challenged by B16F10 cells 2 weeks later. Antitumor efficacy and mechanisms of the vaccine were analyzed. Vaccination with the viable B16F10/ESAT-6-GPI-IL-21 vaccine resulted in an increase of IFN-γ level and the CD8(+)CTL cytotoxicity, a decrease in TGF-β generation and increase in the expression of miR-200c that serves as melanoma suppressor by directly targeting zinc-finger E-box binding homeobox 1 to inhibit epithelial-mesenchymal transition and block tumor metastasis. The vaccine significantly inhibited the melanoma growth, reduced the lung melanoma nodules, and prolonged the mouse survival compared with the controls. These findings highlighted IL-21 as an immune adjuvant in an engineered viable tumor vaccine to reinforce heterogenetic antigen ESAT-6 immune tolerance break to induce powerful antitumor efficacy in mice.
Immunologic Research 04/2012; 52(3):240-9. · 3.03 Impact Factor
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ABSTRACT: The goal of this study was to evaluate the protective efficacy of a cationic nanoparticle-based DNA vaccine expressing antigen 85A (Ag85A) and 6-kDa early secretory antigen target (ESAT-6) of Mycobacterium tuberculosis as well as cytokine interleukin-21 (IL-21) against M. tuberculosis infection. The results of this indicated that the anti-M. tuberculosis immune responses were induced in mice that had received the different DNA vaccines. More importantly, compared with using DNA vaccine Ag85A-ESAT-6-IL-21 alone, the nanoparticle-based DNA vaccine Ag85A-ESAT-6-IL-21 showed a statistically significant increase in the protective efficacy against M. tuberculosis infection in the immunized mice. We concluded that the nanoparticle-based DNA vaccine induced a strong immune response and markedly inhibited the growth of the M. tuberculosis in the mice. These findings highlighted the potential utility of Fe(3)O(4)-Glu-polyethyleneimine nanoparticles encapsulated with the DNA vaccine as a prophylactic vaccine in the M. tuberculosis-infected mouse model. FROM THE CLINICAL EDITOR: This study emphasizes the potential utility of Fe(3)O(4)-Glu-polyethyleneimine nanoparticles encapsulated with DNA vaccine against TB as a prophylactic vaccine. The authors demonstrated a strong immune response and marked growth inhibition of mycobacterium tuberculosis in the mice.
Nanomedicine: nanotechnology, biology, and medicine 03/2012; 8(8):1337-44. · 5.44 Impact Factor
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ABSTRACT: Ovarian cancer causes more deaths than any other cancer of the female reproductive system, and its overall cure rate remains low. The present study investigated human umbilical blood mononuclear cell (UBMC)-derived mesenchymal stem cells (UBMC-MSCs) as interleukin-21 (IL-21) gene delivery vehicles for ovarian cancer therapy in nude mice. MSCs were isolated from UBMCs and the expanded cells were phenotyped by flow cytometry. Cultured UBMCs were differentiated into osteocytes and adipocytes using appropriate media and then the UBMC-MSCs were transfected with recombinant pIRES2-IL-21-enhancement green fluorescent protein. UBMC-MSCs expressing IL-21 were named as UBMC-MSC-IL-21. Mice with A2780 ovarian cancer were treated with UBMC-MSC-IL-21 intravenously, and the therapeutic efficacy was evaluated by the tumor volume and mouse survival. To address the mechanism of UBMC-MSC-IL-21 against ovarian cancer, the expression of IL-21, natural killer glucoprotein 2 domain and major histocompatibility complex class I chain-related molecules A/B were detected in UBMC-MSC-IL-21 and in the tumor sites. Interferon-γ-secreting splenocyte numbers and natural killer cytotoxicity were significantly increased in the UBMC-MSC-IL-21-treated mice as compared with the UBMC-MSCs or the UBMC-MSC-mock plasmid-treated mice. Most notably, tumor growth was delayed and survival was prolonged in ovarian-cancer-bearing mice treated with UBMC-MSC-IL-21. Our data provide important evidence that UBMC-MSCs can serve as vehicles for IL-21 gene delivery and inhibit the established tumor.
Biotechnology and Applied Biochemistry 11/2011; 58(6):397-404. · 1.53 Impact Factor
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ABSTRACT: CSCs (cancer stem cells) are a small subset of cells within a tumour that possesses the characteristics of stem cells and are considered to be responsible for resistance to chemoradiation. Identification of CSCs through stem cell characteristics might have relevant clinical implications. In this study, SP (side population ) cells were sorted from a human ovarian cancer cell line by FACS to determine whether cancer stem cell-like SP cells were present. A very small fraction of SP cells (2.6%) was detected in A2780 cells. SP cells possessed the following characteristics: highly proliferative activity, marked ability for self-renewal in soft agar and culture medium, high expression of ABCG2, drug resistance to vinblastine in vitro, and strong tumourigenic potential in Balb/c nude mice. It is concluded that there exists in the A2780 cell line a small number of SP cells with high expression of ABCG2. The cells have the characteristics of cancer stem-like cells, and identification and cloning of such human SP cells can help in improving therapeutic approaches to ovarian cancer in patients.
Cell Biology International 01/2011; 35(3):227-34. · 1.48 Impact Factor
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ABSTRACT: An apoptotic tumor cell serves as a potential potent trigger for the initiation of naturally occurring tumor immunity. In the present study, a B16F10 tumor cell vaccine treated with mitoxantrone (MIT) was developed, and its antitumor effect on mice was evaluated. The results indicated that the B16F10 tumor cell vaccine treated with MIT alone or in combination with reserpine (RP) and verapamil (VP) for 12 h triggered apoptosis, and that the expression of CD80, the MHC II class molecule, NKG2D and its ligand were significantly increased compared to the expression levels in the control group. The tumor vaccine immunogenicity was significantly enhanced in the vaccinated mice, resulting in augmented cytotoxicity of splenocytes and NK cells as well as the splenocyte proliferative response compared to the control group mice. Notably, the mice vaccinated with the B16F10 tumor cell vaccine treated with MIT, RP and VP did not generate tumors only after 60 days into the observation, but the mice also generated a powerful immune prophylactic efficiency against the B16F10 tumor cell challenge. These findings demonstrated the safety and efficacy of the B16F10 tumor cell vaccine treated with MIT alone or in combination with RP and VP in the murine model, and suggest that an apoptotic tumor cell vaccine modeled on naturally occurring tumor immune responses in vivo may provide a safe and immunogenic tumor vaccine for potential applications in humans.
Experimental and therapeutic medicine 01/2011; 2(5):911-916.
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Weihua Hu, Jing Wang,
Jun Dou,
Xiangfeng He,
Fengshu Zhao,
Cuilian Jiang,
Fangliu Yu,
Kai Hu,
Lili Chu,
Xiaoli Li,
Ning Gu
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ABSTRACT: In the present study, CD34(+) human umbilical cord blood stem cells (UCBSCs) were engineered to express interleukin-21 (IL-21) and then were transplanted into A2780 ovarian cancer xenograft-bearing Balb/c nude mice. The therapeutic efficacy of this procedure on ovarian cancer was evaluated. The findings from the study indicated that UCBSCs did not form gross or histological teratomas until up to 70 days postinjection. The CD34(+) UCBSC-IL-21 therapy showed a consistent effect in the ovarian cancer of the treated mice, delaying the tumor appearance, reducing the tumor sizes, and extending life expectancy. The efficacy was attributable to keeping CD34(+) UCBSC-IL-21 in the neoplastic tissues for more than 21 days. The secreted IL-21 not only increased the quantity of CD11a(+) and CD56(+) NK cells but also increased NK cell cytotoxicities to YAC-1 cells and A2780 cells, respectively. The efficacy was also associated with enhancing the levels of IFN-γ, IL-4, and TNF-α in the mice as well as the high expressions of the NKG2D and MIC A/B molecules in the tumor tissues. This study suggested that transferring CD34(+) UCBSC-IL-21 into the nude mice was safe and feasible in ovarian cancer therapy, and that the method would be a promising new strategy for clinical treatment of ovarian cancer.
Cell Transplantation 11/2010; 20(5):669-80. · 5.13 Impact Factor
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ABSTRACT: In the present study, we developed the tumor vaccine expressing IL-21 in the GPI-anchored form together with secreting GM-CSFs and investigated its antitumor efficacy in C57BL/6 mouse model. The fusion genes containing IL-21 and the GPI anchor signal sequence were acquired by overlaping PCR, inserted into the downstream of two multi-clone sites in recombinant plasmid pRSC/GM-CSFs to form pRSC/IL-21-gpi-GM-CSFs that was transfected into the B16F10 cells. The tumor cell vaccine B16F10/IL-21-gpi-GM-CSFs was identified by reverse transcription PCR, IFA and FCM, respectively. The results showed that the pRSC/IL-21-gpi-GM-CSFs had no cell cycle and proliferative state impact on the B16F10 cells after transfected, and that the tumor vaccine B16F10/IL-21-gpi-GM-CSFs increased the cytotoxicities of NK cells and CD8(+)CTL, enhanced the level of serum IFN-gamma, augmented therapy of tumor effect and prolonged survival time in the tumor-bearing mice immunized with the tumor vaccine B16F10/IL-21-gpi-GM-CSFs. The data that we presented here provided a rationale and practical platform for clinical testing of enhancing cell therapy of B16F10 melanoma efficacy by modified tumor vaccine expressing GPI-anchored IL-21 and secreting GM-CSF.
Vaccine 02/2010; 28(16):2846-52. · 3.77 Impact Factor
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Jun Dou,
Ping Wen,
Weihua Hu,
Yating Li,
Yun Wu,
Chunsheng Liu,
Fengshu Zhao,
Kai Hu, Jing Wang,
Chuilian Jiang,
Xiangfeng He,
Ning Gu
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ABSTRACT: Increasingly more evidence shows that TSCs possess the characteristics of stem-like cells. However, a link between stem cells and TSCs remains to be shown. We have sorted SP cells and non-SP cells from the B16F10 cell lines by FACS, and then studied their cellular biological characteristics by using a SFCM culture method, proliferative assay in vitro, clone formation assays in soft agar and normal media, tumorigenic assays in C57BL/6 mice, and resistance to chemotherapy assay in vitro, the quantitative detecting expression of ABCG2 and their CD phenotype analysis by a FCM. We detected 0.96% SP cells in the B16F10 cells and found that they had obvious differences in characteristics from non-SP cells. They possessed a marked capacity for self-renewal in soft agar and culture medium, strong tumorigenic potential in C57BL/6 mice, apparent resistance to vinblastin in vitro, upregulated ABCG2 expression, and a high expression of CD44+CD133+CD24+ phenotypes. We conclude that there were a few of SP cells that had the characteristics of tumor stem-like cells which may provide a useful tool and a readily accessible source for further study when specific TSCs markers are unknown.
Cell Biology International 06/2009; 33(8):807-15. · 1.48 Impact Factor
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Jun Dou,
Quan Tang,
Fangliu Yu,
Haitao Yang,
Fengshu Zhao,
Weiguo Xu, Jing Wang,
Weihua Hu,
Kai Hu,
Chunsheng Liou,
Xiang Feng He,
Yaqing Wang
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ABSTRACT: Mycobacterium tuberculosis (M. tuberculosis) has once again become a major public health threat owing to the combined effects of a worldwide anti-tuberculosis drug resistance and the emergence of the human immunodeficiency virus pandemic. The Bacille Calmette-Guérin (BCG)-based vaccine has displayed inconsistent efficacy in different trials although it continues to be used to prevent tuberculosis in many countries. In current work, we have developed DNA vaccines expressing M. tuberculosis antigen 85A (Ag85A) and cytokine granulocyte macrophage colony stimulating factor (GM-CSF) and aimed to investigate the immune effect in mice based on BCG priming and DNA vaccine boosting and immune protection against M. tuberculosis challenge. Our results showed that the activity of cytotoxic T lymphocyte and spleen cell proliferative responses to Ag85A and IFN-gamma level as well as the specific antibody titer against Ag85A were significantly increased in mice immunized with prime-boost strategy in comparison with the mice immunized with BCG or DNA vaccine expressing Ag85A and GM-CSF alone. Meanwhile, the immune strategy of BCG-prime and DNA vaccine boost induced mice to generate efficient immune protection against M. tuberculosis challenge. Our data demonstrate that BCG-prime and DNA vaccine expressing Ag85A and GM-CSF boost provides a rational strategy for further development of DNA vaccine against M. tuberculosis infection.
Immunobiology 06/2009; 215(2):133-42. · 3.20 Impact Factor
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Fengshu Zhao,
Jun Dou, Jing Wang,
Lili Chu,
Quan Tang,
Yongfang Wang,
Yating Li,
Minggang Cao,
Weihua Hu,
Kai Hu,
Xiang Feng He,
Ning Gu
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ABSTRACT: GPI-anchored membrane cytokines have been shown to play an important role in host immune response against tumor cells. In the present study, we constructed the tumor vaccine expressing mIL-21 in the GPI-anchored form and investigated its anti-tumor effect in C57BL/6 mice model. The fusion genes containing mIL-21 and the GPI anchor signal sequence was acquired by overlaping PCR, inserted into plasmid pcDNA3.1 to form the pcDNA3.1 mIL-21-GPI recombinant, which was transfected into the B16F10 cells, and the tumor vaccine based on B16F10 cells expressing the GPI-anchored membrane mIL-21 was generated. Through transfection, it was found that GPI-anchored membrane mIL-21 has no proliferate impact on B16F10 cells, but it was functional and reflected in inducing CD3-activated murine splenocytes proliferation response to B16F10 cells, improving the cytotoxicities of CTL and NK cells, increasing the numbers of splenocytes-producing IFN-gamma in mice, augmenting therapeutic effect of tumor and prolonging longevity effects in tumor-bearing mice injected with the inactivated GPI-anchored mIL-21 tumor vaccine. We concluded the expression of mIL-21 on the B16F10 cells surface in the GPI-anchored form was proved to be effective in activating immune responses against tumor cells, and our results provided a good foundation for further investigating the immunotherapy of tumor by GPI-mIL-21.
Immunobiology 06/2009; 215(2):89-100. · 3.20 Impact Factor
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Jun Dou,
Yongfang Wang, Jing Wang,
Fengshu Zhao,
Yating Li,
Minggang Cao,
Weihua Hu,
Kai Hu,
Xiang Feng He,
Lili Chu,
Chuilian Jiang,
Ning Gu
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ABSTRACT: The ovarian cancer cells (SKOV3) secreting IL-21 alone or combination with GM-CSF cytokines was developed and its antitumor effect was evaluated in the nude mice. The gene of IL-21 was amplified from plasmid pRSC-IL-21 by PCR, cloned into the plasmid pRSC-GM-CSF, and the plasmid pRSC-GM-CSF-IL21 was constructed. The plasmids of pRSC-GM-CSF, pRSC-IL21, pRSC-GM-CSF-IL21 and pRSC were respectively transfected into the SKOV3 cells and antitumor efficacy induced by the SKOV3 cells secreting IL-21 or combination with GM-CSF was evaluated by surveying the tumor growth and the nude mice's survival. The results indicated that the secreted IL-21 and GM-CSF were functional because the culture supernatant of SKOV3 cells transfected with the plasmid pRSC-GM-CSF-IL21 enhanced NK cytotoxicity in vitro. The expressions of MIC A/B, NKG2D and ICAM-1 molecules on the SKOV3 cells were up-regulated. The level of IFN-gamma and TNF-alpha, the NK cytotoxicity and the antitumor efficacy were significantly increased in the null mice inoculated with the SKOV3 cells secreting both IL-21 and GM-CSF in comparison with the nude mice inoculated with the other different SKOV3 cells. We concluded that the SKOV3 cells genetically engineered to secrete biologically active IL-21 and GM-CSF elicited antitumor immunity effectively through enhancing NK cytotoxicity, promoting the expressions of MIC A/B , ICAM-1 and NKG2D molecules as well as elevating level of IFN-gamma and TNF-alpha in the nude mice model.
Immunobiology 02/2009; 214(6):483-92. · 3.20 Impact Factor
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ABSTRACT: Automatic exposure (AE) is one of the indispensable functions of modern video cameras. According to the attention mechanism
of human visual systems, peak regions in luminance histogram correspond to the region of no interest in an image. Based on
this assumption, a new AE algorithm using the luminance histogram of an image is proposed in this paper. The algorithm finds
the first two largest peak regions in the histogram and calculates the mean weighted luminance (MWL) of the entire image by
weighting the luminance of pixels inside the two peak regions. The MWL is then used to control the exposure of video cameras.
The weight of pixel luminance is decided by a set of quadratic curves, and the parameters of the quadratic curves are affected
by the brightness of the image background. Fuzzy logic is also applied to optimize the practical AE systems. Results show
that the proposed algorithm gives efficient exposure control over various scene tests.
Frontiers of Optoelectronics in China 01/2008; 1(3):285-291.
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ABSTRACT: The hepatitis C virus (HCV) C region has been reported to have overlapping genes or regions, and may encode a core shadow protein that has a role in HCV self-replication, pathogenesis and carcinogenesis. The aim of this study was to identify the effect of HCV core shadow protein expressed in a human hepatoma (Huh-7) cell line on human gene expression profiles.
Recombinants for expression of HCV genotype 1b core shadow protein and genotype 1b core protein were constructed, and an Huh-7 cell line was established that could express the shadow protein and the core protein constitutively. Affymetrix human gene chip, HG-U133 A and B microarray analysis and semiquantitative RT-PCR were employed to identify the expression profiles of two kinds of core proteins in the Huh-7 cell line.
The microarray analysis showed that the core shadow protein caused expression of more genes to be up/down-regulation than the core protein, including signal transduction, protease activity, molecular transport and, particularly, immune responses genes. Surprisingly, the core shadow protein could increase/decrease expression of apoptosis and anti-apoptosis genes simultaneously. The expression profiles of three up-regulated genes were confirmed by semiquantitative RT-PCR, with results similar to the microarray analysis.
Hepatitis C virus core shadow protein may play an important role in inhibiting or stimulating host cells apoptosis processing and carcinogenesis, which is useful for the understanding of HCV core shadow protein biological functions in vivo and in vitro.
Journal of Gastroenterology and Hepatology 01/2007; 21(12):1794-800. · 2.87 Impact Factor
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ABSTRACT: In present investigation, we constructed recombinants expressing the HCV genotypes 1b, 2a, and 4d core proteins, and established human hepatocellular carcinoma (HepG2) cell line that expressed various genotype core proteins. The gene expression profiles in the cells expressing different HCV genotype core proteins were compared with those in the control by microarray analysis. In data analysis, a threshold was set to eliminate all genes that were not increased or decreased by 2.5-fold change in a comparison between the transfected cells and control cells. The preliminary microarray analysis suggests that the gene expression profiles regulated by three kinds of genotype core proteins are mainly involved in transport, signal transduction, regulation of transcription, protease activity, etc., and that some pathogenesis/oncogenesis gene expressions are up/down- regulated simultaneously in the HepG2 cell line. The data suggest that each core protein has its gene expressions profile and that the profiles are implicated in HCV replication and pathogenesis, which may open up a novel way to understand the function of the HCV variant core proteins biological and their pathogenic mechanism.
Cellular & molecular immunology 07/2006; 3(3):227-33. · 2.99 Impact Factor
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ABSTRACT: To analyze three kinds of genotype hepatitis C virus (HCV) core protein expressed in human hepatoma (Huh-7) cell line and to recognize HCV core proteins biological function and its pathogenic mechanism.
The Huh-7 cell expressed three kinds of core proteins were established respectively. Affymetrix human gene chip was used for identifying the gene expression dependently on Affymetrix's protocol. All genes changed by 3 or 1.5 folds between the transfected cells and a control cells were further analyzed, and annotated by using NetAffx analysis through Affymetrix website and were categorized based on their biological processes.
The HCV-1b core protein caused 16 genes up/down-regulated expression, of which the immune response genes of PF4V1 and SPP1 were up-regulated 3.4 or 4.4 folds respectively. The HCV-2a core protein had caused the immune response gene CXCL5 and apoptosis gene BTF a down-regulated expression of 3.4 and 3.1 folds respectively, but caused the apoptosis genes of HRK and LZTS1 an up-regulated expression of 3.2 and 3.4 folds respectively. As compared with HCV 1b or 2a core protein, HCV-4b core protein caused 111 genes expression changing and it had more obvious effects on gene expression. If we applied 1.5 fold change for a comparison gene expression, a few of the same gene expression profiles might be caused by these two core proteins.
The three kinds of HCV core protein should have its own expression character and be mainly shown in immune responses, signal transduction, apoptosis, etc. It should be helpful for our recognizing the HCV core protein biological function and its pathogenic mechanism.
Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] 02/2006; 40(1):38-41.
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ABSTRACT: To assess GM-CSF immune accessory effects in tumor-bearing mice, an animal tumor model was established by inoculating SP2/0 myeloma cells s.c. into the flank of Balb/c mice and 14 days later, injecting either 400 mug recombinant pcDNA3.1/mGM-CSF or a blank plasmid s.c. or i.m. into the tumor four times. The tumor weight, the activities of CTL and NK, the serum levels of IFN-gamma, IL-2 and lymphocytes infiltrating in tumor tissue were analysed 8 weeks later with MTT, ELISA and pathological section methods. The results showed that the tumor lump was reduced in mice injected s.c. (0.880 +/- 0.405 g) or i.m. (0.378 +/- 0.411 g) with pcDNA3.1/mGM-CSF compared with control mice injected s.c. (1.548 +/- 0.221g, P < 0.01)or i.m. (1.554 +/- 0.249g, P < 0.001) with a blank vector. Lymphocyte infiltration in tumor tissues was very apparent in mice injected i.m. with pcDNA3.1/mGM-CSF. In contrast, there was no lymphocyte infiltration in tumor tissues of control mice. In addition, the serum concentrations of IFN-gamma, IL-2 and the activities of CTL and NK cells were significantly increased in mice injected with pcDNA3.1/mGM-CSF compared with a control mice (P < 0.01). In conclusion, direct gene immunization of recombinant pcDNA3.1/mGM-CSF is a feasible strategy for tumor therapy.
Immunological investigations 02/2006; 35(2):227-37. · 1.16 Impact Factor
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ABSTRACT: To investigate the inhibitory effect of Chinese herbal medicine on the transcription of hepatitis C virus (HCV) structural gene in Hela D cells.
Hela cell line was transfected with recombinant pBK-CMV-HCV containing HCV structural gene by Lipofectamine. RT-nested-PCR and Western blot assay were used to testify the HCV gene expression in Hela cells. The Hela cells expressing HCV structural protein were named Hela D cells. Prescriptions of Xiao chaihu Decoction (XCHD), Fufang Huangqi (FFHQ) and Bingganling (BGL) were respectively added to Hela D cells in various concentrations. Semi-quantitative RT-nested-PCR product analysis was performed according to the fluorescent density between HCV DNA band and GAPDH DNA band in gel electrophoresis after screened.
Recombinant pBK-CMV-HCV could correctly express the HCV structural gene in Hela D cells. After co-culture of Hela D cells with three prescriptional different concentrations for 48 h respectively, the transcription of HCV gene decreased with increasing of the concentration of each prescription. The lightness ratio of HCV product bands to GAPDH product bands was 0.24, 0.10 and 0.12 in Hela D cells incubated with 0.1 g/mL of XCHD, FFHQ and BGL respectively and the lightness ratio HCV product bands to GAPDH product bands was 0.75, 0.67 and 0.61 respectively in the control cells.
The prescriptions of XCHD, FFHQ and BGL partly inhibit the transcription of HCV structural gene in Hela D cells.
World Journal of Gastroenterology 07/2005; 11(23):3619-22. · 2.47 Impact Factor
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ABSTRACT: A novel tuberculosis (TB) gene vaccine containing mouse granulocyte macrophage-colony stimulating factor (mGM-CSF) and a TB antigen (Ag85A) was developed in this study. The genes encoding Ag85A and mGM-CSF were amplified by PCR respectively from the Ag85A-containing pBSby5 and pC-mGM-CSF. The genes were then cloned into two different polylinker sites of plasmid pIRES, forming a novel TB gene vaccine construct pI85AGM. Following transfection of pI85AGM plasmid into 7721 cell line by Lipofectamine(TM), the expression of Ag85A and GM-CSF proteins was identified by Western blotting or RT-PCR. Then Balb/c mice were inoculated with the recombinant pI85AGM, pI85A, pIGM or plasmid alone, respectively. The activities of CTL, NK cells and the Ag85A-stimulated proliferation of spleen cells were measured by MTT method. The serum antibody against Ag85A was detected by ELISA. The results showed that the Ag85A and GM-CSF proteins could be expressed in 7721 cell line and the activity of CTLs and the proliferation of spleen cells were significantly increased in the pI85AGM-immunized mice, indicating that the pI85AGM-immunized mice could generate specific immune responses to Ag85A. This study might provide possibility for developing novel anti-TB gene vaccine.
Cellular & molecular immunology 03/2005; 2(1):57-62. · 2.99 Impact Factor
