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OBJECTIVE: To evaluate the role of total glucosides of peony (TGP) as adjuvant therapy for prevention of cardiac allograft rejection in rats. METHODS: Rats with cardiac allograft were randomly divided into control group, tacrolimus-treated group, TGP-treated group and tacrolimus plus TGP-treated group. Graft survival time was observed. Allografts in some cases were examined by histological study seven days after transplantation. At the same time, the levels of CD4(+) and CD8(+) T cell subsets in peripheral blood were examined by using flow cytometry; the hepatic function and renal function of recipients were also tested. RESULTS: The graft survival time of the tacrolimus-treated group and tacrolimus plus TGP-treated group was (11.14+/-1.57) d and (13.57+/-1.99) d, respectively. The graft survival time of the tacrolimus plus TGP-treated group was longer than that of the tacrolimus-treated group (P<0.05). The histological study showed that the rejection of the tacrolimus plus TGP-treated group was slighter than that of the tacrolimus-treated group. The levels of CD4(+) T cell subset in the peripheral blood of the tacrolimus-treated and tacrolimus plus TGP-treated groups were (38.71+/-5.15)% and (32.43+/-4.39)% respectively 7 days after transplantation. The level of CD4(+) T cell subset in the tacrolimus plus TGP-treated group was lower than that in the tacrolimus-treated group (P<0.05). The level of CD8(+) T cell subset and the hepatic and renal function had no significant differences between the tacrolimus-treated group and the tacrolimus plus TGP-treated group. CONCLUSION: Effects of tacrolimus plus TGP in prevention of rejection are better than tacrolimus monotherapy in rats with cardiac allograft and without increasing side effects.
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Objective To observe the inhibition effect of chitlsan oligosaccharide on tumour. Methods The growth of tumour were observed after chitlsan oligosaccharide was injected into abdominal cavity(ip) or introtumour of mice;And the anti proliferation activities of chitlsan oligosaccharide on cells were evaluated by means of MTT assay. Results Growth of tumour were inhibited by chitlsan oligosaccharide, necrosis was observed in the tumour tissue.Chitlsan oligosaccharide affected the growth of cell. The effect was related to concentration of chitlsan oligosaccharide,but not depending of culture time.The apoptosis of cells could be seen under the transmission electron.Conclusion Chitlsan oligosaccharide could inhibit the growth of tumour and might induce apoptosis of cell.
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Objective:To investigate the effect of human Leptin on the proliferation of human circulating T lymphocytes in vitro and the influence of Leptin on the action of PHA on the proliferation of human circulating T lymphocytes.Methods:PBMC were isolated from hepartinized venous blood of normal donors by density-gradient sedimentation over Ficoll-Hypaque and cultured in the 1640 complete medium.T lymphocytes were then obtained by depleting the monocyte and B cell populations.Cells were then cultured in the 1640 complete medium for 56 h in the presence of PHA.The proliferative effect was assessed by means of incorporation of thymidine.TdR was added and cultured for 16 hours.Cells were then processed in the harvester to measure the incorporation of [()~3H] thymidine.Radioactivity was determined by liquid scintillation counter.Results:The results showed that Leptin alone did not affect the proliferation of T lymphocytes,but it could enhance the effect of PHA on the proliferation of T lymphocytes at the concentrations of PHA from 2-8 ?g/ml.A dose-response studies of T lymphocyte proliferation showed that the maximal effect of Leptin were observed at 10 nmol/L.Leptin did not affect the proliferation of T lymphocytes when the concentrations of PHA were over 8 ?g/ml.Conclusion:The studies demonstrated that Leptin alone did not affect the proliferation of lymphocytes,but it could enhance the action of PHA on cell growth in a dose-dependent manner.
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Objective:To observe mutagenesis of retrovirus and adenovirus as transgenic vector,and provide safe clinic evidence for transgenic tumor cell as tumor vaccin.Methods:Cells were cultured together with virus.Then,DNA and supernatant were carried on an in vestigation in mutagenesis by means of laboratory technique about genetic toxicology.Results:The result indicated that DNA and supernatant of transgenic cell had no mutagenesis through test both In vivo and in vitroConclusion:The virus modified had no mutagenesis as transgenic vector.
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Objective:To study the morphologic and phenotypic characters of a macrophage-tumor vaccine,and to observe the effect of macrophage-tumor vaccine on inducing CTL respose.Methods:The super-structure and the expression of CD14,CD68,CD80,CD86,MHC Ⅱ molecules of macrophage-tumor cells were detected with electron microscope,immunofluorescence staining and flow cytometry respectively.Meanwhile,H22 tumor cells were transplanted to the mice that had been immunized with different tumor vaccines.The weight and volume of tumors,the tumor cell injure rate and the level of LDH in culture supernatant were detected with direct measurement,MTT and selection methods.Results:The macrophage tumor vaccine cells were large cells with an irregular outline,and generally displayed pseudopodium,membrane folding,and vesicles on the cell surface.The predominant cytoplasmic organelles were lysosomes,secondary lysosomes and residual bodies.The percentage of CD14,CD68,CD80,CD86 and MHC Ⅱ positive cells within the differentiated population were 53.90%,98.60%,26.50%,90.20% and 25.40% respectively.The results of experiment in vivo revealed that the tumor forming rate,volume and weight of the group immunized with macrophage-tumor vaccine were much lower than that of control group and the group that were immunized with the macrophages that were induced by liquid paraffin (P0.05),the tumor weight and volume of the group immunized with the macrophage-tumor vaccine were lower than those of the group immunized with inactivated tumor cells(P
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Objective: To study the biological characteristics of tumor cells infected with recombinant adenovirus expressing hTNF-?, investigate the antitumor effect of recombinant adenovirus. Methods: Human lung adenocarcinoma cell line Anip973 was infected with recombinant adenovirus expressing hTNF-?. Cell growth assay, colone formation test, flowcytometry assay and morphology were used to observe the effects on tumor cells. The hTNF-a gene, which was transduced into cancer cells mediated by recombinant adenovirus, was detected by PCR and agarose gel electrophoresis and its products were detected by ELISA assay. The intratumoral injection of rAd-LacZ and rAd-hTNF-? was carried out to evaluate their antitumor effects. Results: The liter of rAd reached 1010 PFU/ml and more than 90% Anip973 cells could be infected by 30MOI rAd. Except the surface structure and ultrastructure of tumor cells infected with rAd had a light change, cell growth abillity assay, colone formation test, flow cytometry assay showed no significant difference compared with that of the control cells. The TNF-? gene expression at 24 h increased greatly. Antitumor study indicated that on the tumor-bearing mice treated with rAd the tumor grew slowly. Tumor volume was significantly smaller and survive time was prolonged than that of controls. Conclusion: There was no significantly changes occurred on tumoral cells after infected with recombinant adenovirus expressing hTNF-?. The intratumoral injection of rAd-LacZ and rAd-hTNF-? could inhibit the growth of solid tumor.
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Objective: To investigate the safety of transgenic human lung adenocarcinoma cell line SPC-A-1/IL-2 as tumor vaccine. Methods: IL-2 gene was introduced into human lung adenocarcinoma cell line SPC-A-1 and was expressed stably on the basis of the construction of retroviral packing cell line PA317/pLIL-2SN.The mutagenesis of both the DNA and supernatant of SPC-A-1/IL-2 in the and in vitro was tested by means of genetic toxicological techniques.Results:The result indicated that mutagenesis of both the DNA and the supernatant of transgenic cell SPC-A-1/IL-2 was not observed. Conclusion: The initial experiment suggested that the application of transgenic cell SPC-A-l/IL-2 as tumor vaccine was bisically safe and reliable.
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Objective: To investigate in vitro the biological characteristics of AdCD80-infected human lung cancer cells on the basis of generation of replication-deficient hB7-1(CD80) recombinant adenovirus. Methods: Human CD80 gene was transduced into lung cancer cells mediated by recombinant adenovirus and then the expression of the gene was detected by PCR and agarose gel electrophoresis. The biological characteristics of the above cells were analysed with electron microscope, FACS and etc. Results: The titers of rAd reached to 10 10 PFU/ml and more than 90% Anip973 lung cancer cells could be infected by 30 MOI rAd. The growth curve and cloning efficiency of rAdCD80-infected Anip973 cells showed no significant difference compared with that of the control cells. The cell proliferation cycle of rAdCD80-infected 973 cells showed no change through FACS test. Having been infected by rAdCD80, the surface structure and ultrastructure of 973 cells had a little change. Conclusion: These results will lay foundation for tumor vaccines.
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In this experiment, we introduced human interleukin-2(lL-2)gene and Neomycin resistance (Neo~R) gene into a human lung adenocarcinoma cell line GLC, using a retroviral vector PLXSN. The positive cells obtained by selecting in G418 were compared with the wild-type cells. We observed their growth characteristics in vitro and the quantity of IL-2 in the culture supernatants. PCR result demonstrated the integration of foreign genes into tumor genome DNA after transfection and permanent existence. Under electron microscope, what can be seen was the surface villi of trans-duced-cells are less and shorter. It deserves further investigation to determine whether the change of cell surface influence metastatic ability of cells.
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Objective:To determine the binding ability of ~V-1 V3 loop to target cells.Methods:V3 loop peptides(V3-HBIO,V3-ADA,V3-89.6)derived from different HIV-1 strains LIIB(X4-nopic),ADA(R5-tmpic),89.6(R5X4-tropic) and the biotinylated V3-BH1O(biotin-BHIO) and V3 -ADA( biotin-ADA) were synthesized. The binding of the biotinylated V3 peptides to cells and the binding targets were analyzed using flow cytometry. Results:V3 BH10 can bind to a wide range of cell lines, while bio-ADA scarcely binds to the monocytes derived from periperal blood mononuclear cells.Antibody anti-CXCR4 binding to cells blocked by V3 -BH10 and biotin-BH1O binding was blocked by protease inhibitors. The binding of V3-BH10 could be significantly enhanced by V3-BHlO but by neither V3-89.6 nor V3-ADA.Conclusion:The binding ability of the V3 loop derived from diverse HIV- I strains are different. The V3 loop derived from HIV-1 X4-tropic strain directly binds to a wide range of cell lines, the binding targets are multiple including at least coreceptor and proteases. The V3 loop derived from R5 -tropic strain ADA does scarcely.
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Objective:To study the biological characteristics of tumor cells infected with recombinant adenovirus expressing hIL 2, and investigate the antitumor effect of the rAd Methods:Human lung adenocarcinoma cell line Anip973 was infected with recombinant adenovirus expressing hIL 2(rAd hIL 2) Cell growth assay, colony formation were used to observe the effects on tumor cells hIL 2 gene was transduced into cancer cells mediated by recombinant adenovirus, then detected by PCR and agarose gel electrophoresis and its products were detected by ELISA assay The intratumor injection of rAd LacZ, rAd hIL 2 was carried out to evaluate its antitumor effects Results:The titer of rAd reached 10 10 PFU/ml and more than 90% Anip973 cells could be infected by 30MOI rAd Growth inhibition assay, colony formation assay showed no significant difference compared with that of the control cells The 24 h IL 2 expression increased greatly Antitumor study indicated that the tumor grew slowly in the bearing tumor mice treated with rAd Tumor volume was significantly smaller and survive time was prolonged than that of controls Conclusion:There were no significantly changes of tumor cells infected with recombinant adenovirus expressing hIL 2 The intratumor injection of rAd LacZ?rAd hIL 2 could inhibit the growth of solid tumor
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Objective:To study the vaccine potency of gene-modified tumor cells. Methods:The EL-4 lymphoma was transduced with recombinant retrovirus containing the murine B7-1 gene. The effect of gene transduction on antitumor immunity was investigated.Results:The appearance, growth rate and surface marker of MHCⅠand MHCⅡ molecules of EL-4 cells transduced with B7-1 gene were the same with control cells except for CD80 positive in B7-1 gene transduced cells. B7-1 gene transduced EL-4 cells resulted in remarkable loss of tumorigenicity in syngenetic mice. EL-4/B7-1 cells could induce system protective immunity. Therapeutic vaccine of EL-4/B7-1 cells could retard the growth of established early-stage EL-4/Wt tumor significantly, but not retard the growth of late-stage EL-4/Wt tumor. Irradiated EL-4/B7-1 vaccine showed weak effect against challenged EL-4 cells.Conclusion:B7-1 gene transduced EL-4 cells can induce system protective immunity. It suggested that this vaccine have a potential application value in human cancer treatment.
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Objective: To investigate the inhibitory effects of chitosan oligosaccharide on tumor growth in vivo. Methods: Murine tumor cell line H22 was inoculated into. Then different doses of chitosan oligosaccharide were injected into the mice-bearing H22 liver carcinoma, IL-2 and IFN-? in the sera were measured by ELISA assay. The in vitro anti-proliferation activity of chitosan oligosaccharide on the lung carcinoma LH-7 cells was evaluated by MTT assay. Results: Chitosan oligosaccharide inhibited in vivo the growth of H22 tumor cells,with increasing the content of IL-2 and IFN-? in sera of the tumor-bearing mice. The weight of spleen and thymus of the mice were increased when compared with those of control group; tissue necrosis was observed in the tumor in situ; chitosan oligosaccharide also inhibited the growth of LH-7 cells in vitro. The inhibitory effect was shown in a concentration dependent pattern, but not correrlated with incubation time. The early characteristics of apoptosis of LH-7 cell could be observed under the transmission electronic microscopy. Conclusion: Chitosan oligosaccharide inhibits proliferation of tumor and improves immunity function of the hosts, and might induce apoptosis of LH-7 cells.