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Objective To investigate the expression of Toll-like receptor 4 (TLR4) and CD80 in unexplained recurrent spontaneous abortion and their relationship,discuss its possible roles in guiding eugenics,and provide a new idea for clinical diagnosis and prevention.Methods The study group were 60 cases with unexplained recurrent spontaneous abortion patients.The control group included 30 cases of normal early pregnancy without previous history of adverse pregnancy.The expressions of TLR4 and CD80 were detected with immunohistochemistry in the peripheral blood,chorion,and decidua from each case.Results TLR4 had higher expression in the peripheral blood,the chorion and decidua from the recurrent spontaneous abortion women than that from normal pregnancy women (P ＜ 0.05).Levels of CD80 were significantly higher in recurrent spontaneous abortion group than in normal pregnancy group (P ＜ 0.05).There was a positive correlation between TLR4 and CD80 (r =0.982,0.986,and 0.776,P ＜0.01).Conclusions The study shows that TLR4 and CD80 were expressed in peripheral blood lymphocytes and villi and decidua of early pregnancy (normal pregnancy or abnormal pregnancy).Higher expressions of TLR4 and CD80 might play an important role in the occurrence of recurrent spontaneous abortion.
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Objective To prepare the mB7-1-GPI-anchored Lewis vaccine and investigate its antitumor effects. Methods mB7-1-GPI was incorporated on Lewis tumor cells and mB7-1-GPI-anchoring tumor vaccine was prepared. The anti-tumor immunity induced by the prepared mB7-1-GPI-anchored Lewis tumor cell vaccine in tumor-bearing mice was observed. Results Flow cytometric analysis showed that mB7-1-GPI were positively expressed on the surface of Lewis tumor cells. After Lewis tumor cells incubated with mB7-1-GPI, the positive rate (PR) of mB7-1 antigen was 95.8% (0h), 93.6% (4h), 91.1% (8h) and the fluorescence intensity (FI) was 11.2(0h), 10. 6(4h), 9. 8(8h). The IL-2 and IFN-γ production of splenic lymphocytes + lewis cells was (25.9 ± 1.4) pg/ml, (56. 0± 3. 5 ) pg/ml. The IL-2 and IFN-γ production of splenic lymphocytes + lewis/mB7-1-GPI was ( 871.3 ± 10. 4 ) pg/ml, ( 1329. 0 ± 11.9 ) pg/ml. In 25 days, the mean diameter of tumor of Lewis/mB7-1 -GPI was shorter than Lewis( 1.4 ± 0. 21 )cm & ( 2. 5 ± 0. 27 )cm , P ＜ 0. 05 ). Lewis tumor cell-bearing C57BL/6 mice treated with Lewis/mB7-1-GPI vaccine survived much longer than mice treated with Lewis vaccine ( 75.2 ± 2. 0 ) d & (40. 2 ± 2. 0 ) d ( P ＜ 0. 05 ). Conclusion The Lewis tumor vaccine prepared with mB7-1-GPI fusion protein significantly inhibited the tumor growth in Lewis bearing mice. It represented an useful new strategy for attaching immunological factor onto tumor cell surfaces without genetic manipulation.
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Objective To observe the effeet of cotransduction of wt-p53,B7-1 and GM-CSF genes mediated by recombinant adenovirus on cell growth, apoptosis and immunogenicity of neuroblastoma cell line SH-SY5Y, Methods We transfected SH-SYSY cells separately with recombinant adenovirus, recombinant adenovirus mediating human wild type p53(wt-p53) gene and BB-102.We separately named them with SH/Ad,SH/p53 and SH/BB-102 group.The protein expression of p53, B7-1 and GM-CSF were measured by Western blot, FCM and enzyme linked immunosorbent assay. Cell number was counted and growth curves were drawed. Cell apoptosis was tested by FCM. Lymphocyte proliferation and cytokine secretory was evaluated by mixed lymphocyte culturing. Results After transection with BB- 102, p53, B7-1, and GM-CSF, high-efficiency expression of target genes were found. Growth of the cells was inhibited, apoptosis was induced in the SH/p53 and SH/BB-102 groups. Proliferation of lymphocytes was stimulated and cytokine increased visorously in the SH/BB-102 group. Condusions After transfection with BB-102, growth of the SH-SY5Y cells was inhibited,apoptosis was induced,and immunogenicity was greatly enhanced.
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Objective To explore the possible role of CD28 +/CD152 +:B7 eostimulators in immune pathophysiology of severe pneumonia.Methods 22 severe pneumonia peripheral blood sample were used to analyze the expression of CD3+ T cell CD28+,CD152+,CD14++ on mononuelear cell CD86+,and HLA - DR + by FACS expression.The relationship between CD28+,CTLA4,CD86+ and the HLA-DR +,and the relationship between APACHE Ⅱ Grading,CD28+,CD152+,CD86+ and HLA-DR + were analyzed.Results Compared with the control group,the expression of CD3 + T cell,CD86+ and HLA - DR + were remarkably reduced while the expression of CD28+ and CD152+ were markedly increased in patients with severe pneumonia who were hospitalized in 24 h(P＜0.05).However,T cells with positive CD8+ CD3+ and CD4+ CD3+ had no significant change between two groups(P＞0.05).For patients with severe pneumonia who survived,the APACHE Ⅱ scores were significantly reduced while the expression of CD28+,CD152+,CD86+,HLA-DR + and CD3+ + cells were significantly increased after 10 days from admission(P＜0.05).By contrast,T cells with positive CD8+ CD3+ and CD4+CD3+ had no significant change between two groups(P＞0.05).There were no relation between costimulators CD28+ and HLA - DR + (r=-0.12,P=0.54)and APACHE Ⅱ scores(r=-0.30,P=0.19) in control group.CD86+ and HLA - DR + showed positive correlation(r=0.65,P=0.00).CD86+ and APACHE Ⅱ scores had no correlation(r=-0.38,P=0.09).Conclusion The costimulators expressed abnormally in circumference blood of patients with severe pneumonia,CD86+ decreased,but CD28+,CD152+ increased.T cell of circumference blood was at the condition of "anergy".The increase of CD28+,CD86+,CD86+ and HLA - DR + during convalescence stages in patient with severe pneumonia showed that spocific immunity was advantageous for restoration in these patients.The relationship among CD86+,CTLA4 and HLA - DR + indicated that CD28+/CD152+:B7 play an role in the occurrence and development of severe pneumonia.
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Objective To study the inhibitory effects of gene combined radiotherapy on mice transplanted with B16 melanoma.Methods Alkaline lysis assay was used to extract and purify the plasmid.Plasmid DNA was injected into tumor by microinjection assay.Mice were inoculated with 5?10~5 of B16 melanoma in right hind legs and the therapy was performed when the diameter of tumor reached at 0.30.5 cm.pNE-mIL-12 and pcDNA-B7-1 plasmids were injected locally three times following with irradiation three times.The tumor growth rate of mice was observed.Results The anti-tumor effect of pNE-mIL-12 combined with pcDNA-B7-1 plasmid following with 2 Gy X-ray irradiation was much better than other groups.It showed that the tumor growth rate was slowed,the survival days of mice were delayed significantly(P
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Objective To explored the role of CD40/CD40L and B7-1/CD28 interaction in the immunopathologic process of folic acid-induced nephropathy(FAN) and investigate the intervention effect of the anti-B7-1 monoclone antibody(B7-1mAb). Methods GD1 mice were administrated by i. p with FA only or co-treated with B7-1 mAb for the intervention study. Kidneys were harvested for immunohistochemical, Western blot assays and serum for renal function evaluation at day 1, 3,5, 7, 14 and 21. Results Continuous expression of CD40, B7-1 from day 1 to day 21 on the renal tubular epithelial cells was detected in this model. CD40 was up-regulated more than five times in the kidney at every test time point (P
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50 days),whereas third party skin grafts were vigorously rejected.Conclusion Blockade of CD28-B7 and CD40-CD40 ligand interactions resulted in long-term allograft survival and donor-specific tolerance in mouse cardiac transplantation without chronic rejection.
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Objective To investigate the role of costimulatory molecules CD28/B7 in the pathogenesis of psoriasis. Methods The expression of CD28, CD80 and CD86 was detected in the lesional skin of 22 cases by immunohistochemical technique (ABC), and in the peripheral blood lymphocytes of 17 cases by flow cytometry respectively. As control, normal skin and lymphocytes from peripheral blood of healthy volunteers were also tested. Results The immunohistochemistry study showed that the expression of CD28, CD80 and CD86 in the psoriatic lesions was significantly higher than that in the normal controls (P .05). Conclusion CD28, CD80 and CD86 might play a certain role in the pathogenesis in psoriasis.
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Objective To investigate the significance of B7-CD28/CTLA-4 (CD152),co-stimulatory molecules of T lymphocytes,in the onset,development and prognosis of genital congdyloma acuminatum (CA) in females.Methods Flow cytometry was utilized to detect the expression levels of CD80/CD86 in peripheral blood lymphocytes and of CD28/CD152 (CTLA-4) in CD4~+/CD8~+ T lymphocytes from 30 CA patients (17 primary CA,13 recurrent CA,15 at recovery stage of CA) and 15 healthy volunteers as con- trols.Results No significant difference was found for the frequencies of CD80~+,CD86~+,CD4~+/CD28~+ and CD8~+/CD28~+ lymphocytes between the primary or recurrent group and the control group.The frequencies of CD8~+/CD28~+,CD4~+/CD28~+ and CD80~+ lymphocytes were significantly higher in the recovery group than those in the recurrent group (P0.05).Conclusions There is an abnormal expression of co-stimulatory molecules B7-CD28/CTLA-4 (CD152) in periphoral blood lym- phocytes,CD4~+ and CD8~+ T lymphocytes in female patients with genital CA,and the expression abnormaility is closely linked with different disease stages of CA.
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Objective To investigate the proliferation and cytotoxicity of cytotoxic T lymphocyte (CTL) induced by ovarian cancer cells transfected with a CD 80 gene containing retroviral vector Methods The ovarian cancer cell line TYK cells were transfected with retro virus vector PLXSN hCD 80 After geneticin (G418) selection, the CD 80 expression of the transfectants was confirmed by flow cytometry CTL was induced by co culture of CD 80 expressing TYK cells (TYK hCD 80 ) and peripheral blood mononuclear cells (PBMC) in the presence of anti CD 3 monoclonal antibody (McAb) Proliferation of PBMC was measured using 3?H Thymidine incorporation assay The lysis activity of CTL toward tumor cells was determined using methyl thiazolyl tetrazolium (MTT) assay Results After transfection and G418 selection, the CD 80 expression was proved by flow cytometry The highest rate of CD 80 expression was 84 9% The TYK cell line expressing high CD 80 was named TYK hCD 80 In the presence of anti CD 3 McAb,TYK hCD 80 significantly enhanced proliferation of PBMC than TYK cells ( 3H Thymidine incorporation, (40 604? 842) vs (8 000? 594) cpm ( P
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ObjectiveTo investigate the expression of costimulatory molecules, Fas/FasL, and major histocompatibility complex (MHC)-class Ⅱ antigens in normal ocular tissues.MethodsTwelve eyes were obtained from the eye bank of Zhongshan Ophthalmic Center within 16 to 24 hours postmortem. Six eyes were used for making the retinal wholemounts, and the tissues (iris and ciliary body, choroid, and retina) of the others were used for making the frozen sections. Immunohistochemical staining was carried out on these retinal wholemounts as well as on tissue sections to investigate the exprenion of B7-1 and B7-2 (costimulatory molecules), HLA-DR(MHC class Ⅱ), CD68 (macrophages), Fas/FasL.ResultsThe results of immunohistochemical staining showed the presence of B7-2, FasL, CD68 and HLA-DR in the iris and ciliary body. Expression of B7-1, FasL, CD68, and HLA-DR was found in the choroid while HLA-DR, CD68 and FasL were detectable in the retina.ConclusionExpression of costimulatory molecules, MHC-class Ⅱ molecules and molecules related to apoptosis is different in the iris, ciliary body, choroid, and retina, which may play an important role in the stability of the immunological microenvironment of these tissues.
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Objective To investigate the effects of ?-interferon (IFN-? on the expression of costimulatory molecules B7-1 (CD80) and B7-2 (CD86) and Fas/FasL in human retina. Methods Nine human eyes were obtained from the eye-bank of Zhongshan Ophthalmic Center. Six eyes were used for making retinal wholemounts, and 12 retinal wholemounts from each eye were put into the 24-hole culture board which had 2ml DMEM/F12 culture medium in each hole. The wholemounts were divided into 3 groups whose concentration of IFN-? was 0, 200, and 1 000 U/ml respectively. After cultured in 37℃ culture box (95%O2,5%CO2) for 24 hours, the expressions of B7-1 (CD80), B7-2 (CD86), and Fas/FasL on these retinal wholemounts were detected by immunohistochemical method. The retinal wholemounts from 3 healthy people were detected by immunohistochemical method as the control. Results Expression of FasL but not B7-1, B7-2 or Fas was found in the control group, while the expression of B7-1, B7-2 and Fas and increased expression of FasL were found after cultured with IFN-?. Conclusion IFN-? may be involved in the occurrence of ocular immune response and induction of apoptosis via the stimulation of expression of costimulatory molecules and Fas/FasL, which plays an important role in the activation of T lymphocytes.
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Objective To study the role of the chimeric hsp70/CD80 DNA vaccine for treatment for TB. Methods C57BL/6N mice challenged with H37Rv were immunized with the chimeric hsp70/CD80 DNA vaccine, hsp70 DNA vaccine and BCG in order to compare the therapeutic role of these vaccines. Results The levels of interferon ? (IFN-?) in the serum of mice in hsp70/CD80 group (1336.98?129.64) pg/ml was significantly higher than in group BCG (121.54?56.39) pg/ml, pcDNA3 (192.00?64.36) pg/ml and pcDNA3 hsp70 (542.33?99.77) pg/ml. The growth of Mycobacterium tuberculosis in live and spleen were inhibited significantly in hsp70/CD80 vaccinated mice. Conclusions The chimeric hsp70/CD80 DNA vaccine was more efficient than BCG and hsp70 DNA vaccine alone in treating TB.
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Objective To investigate the expression of costimulatory molecules CD80and CD86on peripheral blood lymphocytes(PBLC)in patients with systemic lupus erythematosus(SLE)and its significance.Methods The expression of CD80and CD86on PBLC was measured by flow cytometric assay in30patients with SLE and25normal controls.Results Expression of CD86on PBLC in patients with SLE was significantly lower than that of normal controls(P