-
[show abstract]
[hide abstract]
ABSTRACT: To study the immunogenicity of the therapeutic multi-epitope gene vaccine of Hepatitis B virus.
Multi-epitope gene of Hepatitis B virus was cloned into prokaryon expression vector pBAD/gIIIA to express a non-fusional antigen B-BPT and BALB/c mice were immunized with the antigen. Cytotoxic T-lymphocyte(CTL) was determined with CytoTox96 kit which quantitatively measures the lactate dehydrogenase(LDH) that is released on cell lysis. CD4(+);, CD8(+); T lymphocytes subsets in immunized mice was detected by using flow cytometry (FCM). Lymphocyte proliferation response was completed by MTT colorimetry.
Injection of B-BPT elicited high-level of CTL response and also stimulus spleen lymphocytes to proliferate effectively.
B-BPT can induce specific cellular immune responses and may be a good candidate for therapeutic vaccine.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 03/2011; 27(3):260-2.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the effects of polyinosinic-polycytidylic acid (polyI:C) on the production of thymic stromal lymphopoietin (TSLP) and airway inflammation in mice with exacerbated asthma induced by respiratory syncytial virus (RSV).
Thirty-two female BALB/c mice were randomly divided into 4 groups, namely the PBS control group, OVA group, OVA/RSV group, and OVA/RSV/polyI:C group. In the latter 3 groups, the mice were sensitized by OVA and stimulated with nebulized OVA. RSV was inoculated into the nasal cavity of the sensitized mice and polyI:C (1 mg/kg) was intramuscularly administered. The airway response to metacholine was examined, and the serum levels of IL-4, IL-5, IL-13, and IFN-γ and TSLP in the supernatants of bronchoalveolar lavage fluid (BALF) were detected using ELISA. The total BALF cells, eosinophils, lymphocytes and neutrophils were counted. The lung specimens were collected to observe the inflammation with HE staining, and immunohistochemistry was employed to determine TSLP production in the airway epithelial cells.
The mice in RSV/OVA/polyI:C group showed a significantly lower airway responsiveness to metacholine than those in OVA/RSV group (P<0.01). Compared with OVA/RSV group, RSV/OVA/polyI:C group showed significantly lower serum levels of IL-4, IL-5, IL-13 and TSLP in BALF (P<0.05), with also lower total BALF cells, eosinophils and lymphocytes (P<0.05) and lessened infiltration of the airway inflammatory cells. Immunohistochemistry of TSLP also demonstrated a lower production of TSLP in the airway epithelial cells in RSV/OVA/polyI:C group than in OVA/RSV group.
polyI:C can inhibit the increase in TSLP production in the airway epithelial cells after RSV infection and relieve airway inflammation in mice with RSV-induced asthma exacerbation.
Nan fang yi ke da xue xue bao = Journal of Southern Medical University 03/2011; 31(3):434-7.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the effects of different factors on the expressions of thymic stromal lymphopoietin (TSLP) in respiratory syncytial virus (RSV)-infected human airway epithelial cell line 16HBE cells.
RSV amplified by infecting Hep-2 cells was identified for its virulence. 16HBE cells were divided into six groups, namely the control group, RSV group, RSV/anti-TLR3 group, RSV/IFN-gamma group, RSV/IL-4 group and RSV/dexamethasone group with corresponding treatments. Real-time RT-PCR was used to examine the expression of TSLP mRNA in the cells 6 h after RSV infection. Western blotting was used to examine TSLP protein expression in the cells 24 h after the infection.
The expression of TSLP mRNA in 16HBE cells 6 h after RSV infection increased by 1.63-/+0.08 folds as compared to the expression level in the control cells. The expression of TSLP mRNA was significantly decreased in RSV-infected cells treated with anti-TLR3 antibody (P=0.034) and recombinant human IFN-gamma (P<0.001), but increased with the treatment by recombinant human IL-4 (P=0.025). Dexamethasone significantly inhibited the expression of TSLP mRNA in RSV-infected cells (P<0.001). The production of TSLP protein in 16HBE cells increased by 1.9 folds (P<0.001) 24 h after RSV infection, but underwent no significant changes after treatment with anti-TLR3 antibody (P=0.114). Recombinant human IFN-gamma significantly decreased while IL-4 enhanced the expression of TSLP protein in the infected cells (P=0.020 and 0.014, respectively). Dexamethasone significantly inhibited the increment of TSLP protein expression in RSV-infected cells (P<0.001).
RSV infection can enhance the expressions of TSLP in human airway epithelial cells. IFN-gamma, anti-TLR3 and dexamethasone can inhibit the elevation of TSLP expression induced by RSV infection, but IL-4 synergistically enhances its expression.
Nan fang yi ke da xue xue bao = Journal of Southern Medical University 03/2010; 30(3):519-22.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the role of ribavirin (RIB) in treating respiratory syncytial virus (RSV)-induced asthma exacerbation of mice.
(1) Cell experiment: 32 flasks of human airway epithelial cell 16HBEs were randomly divided into four groups: RSV group, RSV/RIB group, RIB group and control group. 16HBEs were infected with RSV at a multiplicity of infection (MOI) of 2. RIB 50 microg/ml was added in culture medium and Western blot used to detect the production of thymic stromal lymphopoietin (TSLP) protein; (2) Animal experiment: 32 female BALB/c mice were randomly divided into four groups: Ovalbumin (OVA) group, OVA/RSV group, OVA/RSV/RIB group and control group. Mice were sensitized by OVA and stimulated with nebulized OVA. RSV was inoculated into murine nasal cavity and RIB 10 mg/kg intramuscularly administered. BUXCO noninvasive murine lung function detection instrument was used to examine the airway response to metacholine; ELISA was used to detect IL-4, IL-5, IL-13 and IFN-gamma in murine serum and TSLP in supernatants of bronchoalveolar lavage fluid (BALF); murine lung specimens were stained with HE to observe inflammation and immunohistochemical technique was employed to observe the production of TSLP in murine airway epithelial cells.
The cell experiment demonstrated the productions of TSLP protein in RSV group, RSV/RIB group, RIB group and control group were 1.97 +/- 0.22, 1.16 +/- 0.19, 0.99 +/- 0.17 and 0.89 +/- 0.08 respectively, and the production of TSLP in RSV/RIB group was lower than that in RSV group (P < 0.01). The animal experiment demonstrated that the murine airway responsiveness in RSV/OVA/RIB group was lower than that in OVA/RSV group (P < 0.01). The levels of IL-4 [(109.7 +/- 41.9) pg/ml], IL-5 [(220.8 +/- 30.9) pg/ml], IFN-gamma [(13.0 +/- 3.4) pg/ml] in murine serum and TSLP [(1945 +/- 82) pg/ml] in BALF of RSV/OVA/RIB group were significantly lower than those in OVA/RSV group [(274.2 +/- 103.7), (293.3 +/- 46.1), (30.1 +/- 5.7) and (2127 +/- 46) pg/ml respectively, all P < 0.01]; less infiltration of airway inflammatory cells in OVA/RSV/RIB group was observed than that in OVA/RSV group. Immunohistochemical staining of TSLP also showed a lower production of TSLP in airway epithelial cells of OVA/RSV/RIB group than OVA/RSV group.
Ribavirin can inhibit the elevated production of TSLP after RSV infection and relieve RSV-induced asthma exacerbation in mice.
Zhonghua yi xue za zhi 06/2009; 89(20):1430-4.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the effect of respiratory syncytial virus (RSV) infection on the production of thymic stromal lymphopoietin (TSLP) and Th1/Th2 balance in asthmatic mice.
Thirty-two female BALB/c mice were randomly divided into 4 groups, namely the PBS group, ovalbumin (OVA) group, RSV group and OVA/RSV group. The mice were sensitized by OVA and then stimulated with nebulized OVA, and RSV was inoculated into the nasal cavity of the mice. BUXCO noninvasive lung function detection was performed to examine the airway response to metacholine, and enzyme-linked immunosorbent assay (ELISA) was used to detect the serum levels of IL-4, IL-5, IL-13, and IFN-gamma in the mice. The cells in the bronchoalveolar lavage fluid (BALF) were counted and classified, and the supernatants of the BALF were used for the detection of TSLP. Histopathological changes in the lung tissues of the mice were examined using HE staining, and immunohistochemistry using anti-mouse TSLP antibody was performed to examine TSLP expressions in the airway epithelial cells.
RSV infection promoted the production of TSLP in the asthmatic mice, and the concentration of TSLP in OVA/RSV group (2.13-/+0.05 ng/ml) was significantly higher than that in the other groups (P<0.01). RSV infection increased the serum levels of IL-4, IL-5, IL-13, and IFN-gamma in the mice. The total BALF cells, eosinophils, lymphocytes and neutrophils in OVA/RSV group were significantly higher than those in the other groups; noninvasive lung function examination showed higher Penh value in OVA/RSV group (318.66-/+50.87) than in the other groups when the inhaled metacholine increased to 6.25 mg/ml (P<0.01). More obvious and extensive airway inflammatory cell infiltration in OVA/RSV group were observed, and immunohistochemical staining also showed higher expression of TSLP in the airway epithelial cells of OVA/RSV group.
RSV infection promotes the production of TSLP in the airway epithelial cells and increases the level of Th2 cytokines in asthmatic mice. Concurrent RSV infection can exacerbate Th2 inflammatory reaction in asthmatic mice.
Nan fang yi ke da xue xue bao = Journal of Southern Medical University 04/2009; 29(4):724-8.
-
Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue 03/2009; 21(2):121-2.
-
[show abstract]
[hide abstract]
ABSTRACT: To explore the method of adjusting the immunosuppressants in serious infection after liver transplantation.
With reference to sepsis-related organ failure assessment (SOFA), 2005.1-2007.12, when the patient's score > or =15, the immunosuppressants were withdrawn, and the patients were given powerful antibiotics and the other treatments in combination. They were further divided into two groups, SOFA 15-17 (group A, 10 cases) and > or =18 (group B, 16 cases). They were compared, and also with the patients without stoppage of immunosuppressants (group C, 13 cases, 2003.3-2004.12). After withdrawing the immunosuppressant, the rejection incidence and times, the changes in SOFA score and mortality and their relationships were analyzed.
After adjusting the immunosuppressant and with control of serious infections, rejection occurred in 9 patients, with 5 cases in group A (50.0%), 4 in B (25.0%), none in C. The differences among groups showed statistically significant difference (chi(2)=8.0, P=0.02), but no difference was seen between group A and B (chi(2)=1.70, P=0.19). When the rejection developed, the SOFA score decreased obviously (9.78+/-3.14 vs. 17.22+/-1.86, t=6.10, P=0.00). The time of rejection was (17.56+/-2.60) days after stopping the immunosuppressant. All 25 deaths were due to serious infection with multiple organ dysfunction syndrome, but not rejection. Five deaths occurred in group A (50.0%), 7 in B (43.8%), 13 in C (100.0%). Not a single patient with rejection died from infection. Proper adjustment of the immunosuppressants could decrease the mortality (chi(2)=7.60, P=0.02).
SOFA score could be used to guide the adjustment of the immunosuppressants, when SOFA> or =15, the immunosuppressants could be stopped, which would not increase the rejection incidence and decrease mortality. The lower the SOFA score is, the faster the patients recuperate better, but more rejection develops. In order to adjust the immunosuppressant in time, the period with high SOFA score should be shortened.
Zhongguo wei zhong bing ji jiu yi xue = Chinese critical care medicine = Zhongguo weizhongbing jijiuyixue 02/2009; 21(2):85-8.
-
[show abstract]
[hide abstract]
ABSTRACT: To study the cloning, expression and antigen of therapeutic multi-epitope gene of hepatitis B virus.
The multi-epitope gene of hepatitis B virus(BPT) was designed, synthesized and cloned into the prokaryotic expression vector pBAD/gIIIA. Then it was transformed into E.coli Top10 and multi-epitope protein of hepatitis B virus(B-BPT) was expressed under the induction of Arabinose. The immunogenicity of the protein was analyzed by Western blot detection.
The recombinant plasmid pBAD/BPT was constructed successfully and the protein of multi-epitope gene of hepatitis B virus was expressed in E.coli. Western blot detection showed the protein had ideal antigenicity.
The design of therapeutic multi-epitope gene of hepatitis B virus is proved to be correct. The expressed protein may be a good therapeutic vaccine.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 09/2008; 24(8):788-90.
-
[show abstract]
[hide abstract]
ABSTRACT: To study the immunogenicity of a multiple epitope DNA vaccine against hepatitis B virus(HBV).
A multiple epitope HBV antigen gene BPT was synthesized and cloned into eukaryotic expression vector pcDNA3.1 and then BALB/c mice were immunized with the DNA vaccine. The specific humoral and cellular immune responses were detected by indirect ELISA, cytotoxicity of CTL, and lymphocyte proliferation. The immunized mice were also observed for the possible toxicity and side effects after administration of the DNA vaccine.
Immunization with pcDNA3.1/BPT elicited high-level antigen-specific IgG and antigen-specific CTL response, and stimulated lymphocyte proliferation. RT-PCR analysis of spleen lymphocytes showed that levels of IL-12 mRNA in immunized mice were notably higher than that in control mice.
The multiple epitope HBV DNA vaccine can induce specific humoral and cellular immune responses, which lays a certain foundation for development of prophylactic and therapeutic HBV vaccine.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 10/2004; 20(5):517-21.
-
[show abstract]
[hide abstract]
ABSTRACT: To study the synthesis, cloning, expression and antigenicity of therapeutic multi-epitope gene of hepatitis B virus.
The therapeutic multi-epitope gene of hepatitis B virus was synthesized and cloned into the vector pWR450-1, then was expressed in E.coli and the products were purified. The immunogenicity of the expressed protein was analyzed by Western-blotting.
Recombinant plasmid PWR/BPT was constructed successfully and the protein of multi-epitope gene of hepatitis B virus was expressed in E. coli, which showed ideal antigenicity by Western-blotting.
The designed multi-epitope therapeutic gene of hepatitis B virus was proved to be correct and the expressed protein may be a good therapeutic vaccine.
Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA 09/2003; 23(8):802-5.
-
[show abstract]
[hide abstract]
ABSTRACT: To study the inhibitory effect of recombinant human endostatin (rhES) on the angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795.
The recombinant yeast strain containing the gene sequence encoding highly soluble rhES was induced by methanol for rhES production, which was purified with heparin affinity chromatography. T739 mice with subcutaneous inoculation of LA795 cells were randomized into 2 groups (10 in each group) to receive injection of either rhES (20 mg/kg x b x w x per day) or PBS in the same volume for 14 consecutive days starting from the sixth day after the inoculation. The angiogenesis and lung metastasis of the implanted tumors were subsequently observed.
Purified rhES was successfully obtained. As shown by immunohistochemistry, the tumors in the mice receiving rhES treatment exhibited less density of the microvessels than those in the PBS-treated mice did (P<0.01). Pathological examination of the lung tissue of the mice in rhES group found no visible signs of tumor metastasis, which, in contrast, was widespread in PBS group. The weight of the lungs was also significantly different (P<0.01).
rhES possesses good biological properties and can potently inhibit the angiogenesis and lung metastasis of mouse lung adenocarcinoma LA795.
Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA 01/2003; 23(1):30-3.
-
[show abstract]
[hide abstract]
ABSTRACT: Generally, growth and metastasis of tumor are critically dependent on angiogenesis. Endostatin can specifically inhibits tumor angiogenesis. The current study was designed to evaluate the inhibitory effect of recombinant human endostatin (rhES) secreted by pichia, pastoris, GS115 on the growth and metastasis of mice lung adenocarcinoma LA795 in mice T739.
To select a strain that could highly express recombinant human endostatin, then induce the clone to express rhES by adding methanol. Purification of rhES was performed with heparin affinity chromatography. LA795 tumor cells were inoculated subcutaneously into the dorsa of T739 mice, and the mice were randomized into two groups. The first group was given rhES(20 mg/kg/d), and the second group was given equal volume of PBS, for 14 consecutive days. The volume of tumors were measured. And the tumor metastasis in the lungs of the mice was observed.
The selected clone was induced to secrete enough soluble rhES. The purified protein could strongly inhibit growth and metastasis of mice lung adenocarcinoma LA795 in T739 mice (P < 0.001).
The rhES secreted by pichia, pastoris, GS115 has good biological activities and greatly inhibit growth and metastasis of mice lung adenocarcinoma LA795 in mice T739.
Ai zheng = Aizheng = Chinese journal of cancer 11/2002; 21(11):1197-202.
-
[show abstract]
[hide abstract]
ABSTRACT: To investigate the inhibitory effects exercised by endostatin on the production of interleukin-6 (IL-6) and IL-8 by human umbilical vein endothelial cells (HUVECs).
(HUVECs were isolated and cultured in vitro with endostatin (treated group) or PBS (control group), and the supernatant was harvested from the primary culture medium daily for 9 consecutive days starting from the first day of culture, followed by centrifugation. IL-6 and IL-8 contents in the supernatant were measured using sandwich enzyme-linked immunosorbent assay (ELISA).
IL-6 and IL-8 were detected in the supernatant of the control cell culture, and their amounts increased as the cell culture was prolonged, reaching the peak levels on day 6 (2 979.32+/-19.65 pg/ml and 6 018.87+/-56.74 pg/ml, respectively). In the treated group, however, the amounts of IL-6 and IL-8 were significantly lower than the control levels (P<0.01).
Endostatin can inhibit the growth and proliferation of endothelial cells, reducing their biological activities.
Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA 01/2002; 22(1):54-6.
