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Volume 6,Issue 2,1999 Table of Contents
1999, 6(2):83-86.
DOI: 10.3872/j.issn.1007-385X.1999.2.002
Abstract:
To observe the expression of human wild type p53, B7 1 and GM CSF genes mediated by recombinant adenovirus and their effects of inducing apoptosis on lung cancer cells. Methods: Human wild type p53, B7 1 and GM CSF genes were transfected into lung cancer cells mediated by recombinant adenovirus. The expression products of these genes were detected by immunohistochemistry assay, flow cytometric analysis and ELISA. Cell growth assay was carried out by counting alive cells after trypan blue exclusion. Cell apoptosis was detected by TdT assay of DNA fragmentation. Results: The multi genes could be efficiently expressed in lung cancer cells mediated by recombinant adenovirus, which could suppress lung cancer cell growth and induce their apoptosis. Conclusion: These results suggest the feasibility of muti gene therapy for lung cancer.
To observe the expression of human wild type p53, B7 1 and GM CSF genes mediated by recombinant adenovirus and their effects of inducing apoptosis on lung cancer cells. Methods: Human wild type p53, B7 1 and GM CSF genes were transfected into lung cancer cells mediated by recombinant adenovirus. The expression products of these genes were detected by immunohistochemistry assay, flow cytometric analysis and ELISA. Cell growth assay was carried out by counting alive cells after trypan blue exclusion. Cell apoptosis was detected by TdT assay of DNA fragmentation. Results: The multi genes could be efficiently expressed in lung cancer cells mediated by recombinant adenovirus, which could suppress lung cancer cell growth and induce their apoptosis. Conclusion: These results suggest the feasibility of muti gene therapy for lung cancer.
1999, 6(2):87-90.
DOI: 10.3872/j.issn.1007-385X.1999.2.003
Abstract:
To clone the full length cDNA of the tumor rejection gene MAGE-1 from hepatocellular carcinoma(HCC) tissues. This MAGE-1 gene and the tumor rejection antigen encoded by it may be useful in subsequent studies aiming at exploring new strategies for the immunotherapy for HCC. Methods: The full length MAGE-1 cDNA was amplified by RT-PCR method using a pair of primers designed according to the encoding sequence of MAGE-1 gene. The PCR products were then digested by restriction endonucleases and inserted into the plasmid PUC19. After primary selection of the recombinants by endonuclease digestion, the sequences of the inserted gene fragments were confirmed by DNA sequence analysis. Results; Using the same pair of primers, we obtained three clones of different MAGE genes, which were a full length MAGE-1 gene, a 750 bp fragment of MAGE-3 gene and a gene highly homologous to MAGE-6 and MAGE-12 but not identical to any reported MAGE genes. Conclusion: These data suggested that some MAGE genes are expressed in heptocellular carcinoma probably including some unknown genes, which might introduce potential new targets for immune attacks.
To clone the full length cDNA of the tumor rejection gene MAGE-1 from hepatocellular carcinoma(HCC) tissues. This MAGE-1 gene and the tumor rejection antigen encoded by it may be useful in subsequent studies aiming at exploring new strategies for the immunotherapy for HCC. Methods: The full length MAGE-1 cDNA was amplified by RT-PCR method using a pair of primers designed according to the encoding sequence of MAGE-1 gene. The PCR products were then digested by restriction endonucleases and inserted into the plasmid PUC19. After primary selection of the recombinants by endonuclease digestion, the sequences of the inserted gene fragments were confirmed by DNA sequence analysis. Results; Using the same pair of primers, we obtained three clones of different MAGE genes, which were a full length MAGE-1 gene, a 750 bp fragment of MAGE-3 gene and a gene highly homologous to MAGE-6 and MAGE-12 but not identical to any reported MAGE genes. Conclusion: These data suggested that some MAGE genes are expressed in heptocellular carcinoma probably including some unknown genes, which might introduce potential new targets for immune attacks.
1999, 6(2):91-96.
DOI: 10.3872/j.issn.1007-385X.1999.2.004
Abstract:
To construct the recombinant adenovirus encoding antisense cyclin D1 fragment, and to investigate its effect on tumor cell proliferation and apoptosis. Methods: The shuttle plasmid encoding antisense cyclin D1 was constructed by cloning cyclin D1 cDNA fragment in the reverse direction into the pAdCMV. Then the plasmid pJM17 and the shuttle plasmid were co-transferred into 293 cells with liposome for homologous recombination to acquire recombinant adenovirus. The cell cycle, apoptosis, and the expression of the related genes of the in vitro transferred human adenocarcinoma cell lines GLC-82 and SPC-A-1, as well as the human embryonic diploid lung cell line 2BS were studied. The effect on colony formation was also studied. Results: The recombinant adenovirus encoding antisense cyclin D1 fragment Ad-AS cyclin D1 was obtained with the titer of 1×10 10 pfu/ml. RT-PCR assay showed that the expression of cyclin D1 of GLC-82 and SPC-A-1 was reduced after Ad-AS cyclin D1 infection. Simultaneously, obvious G1 arrest and apoptosis, bcl-2 and bax gene expression changes were observed in the infected tumor cells. The infected 2BS cells showed no such changes. The colony formation ability in vitro was also reduced.]Conclusion: The reduced expression of cyclin D1 gene could induce human lung adenocarcinoma cells specific G1 arrest and apoptosis, while has no effect on normal cell.
To construct the recombinant adenovirus encoding antisense cyclin D1 fragment, and to investigate its effect on tumor cell proliferation and apoptosis. Methods: The shuttle plasmid encoding antisense cyclin D1 was constructed by cloning cyclin D1 cDNA fragment in the reverse direction into the pAdCMV. Then the plasmid pJM17 and the shuttle plasmid were co-transferred into 293 cells with liposome for homologous recombination to acquire recombinant adenovirus. The cell cycle, apoptosis, and the expression of the related genes of the in vitro transferred human adenocarcinoma cell lines GLC-82 and SPC-A-1, as well as the human embryonic diploid lung cell line 2BS were studied. The effect on colony formation was also studied. Results: The recombinant adenovirus encoding antisense cyclin D1 fragment Ad-AS cyclin D1 was obtained with the titer of 1×10 10 pfu/ml. RT-PCR assay showed that the expression of cyclin D1 of GLC-82 and SPC-A-1 was reduced after Ad-AS cyclin D1 infection. Simultaneously, obvious G1 arrest and apoptosis, bcl-2 and bax gene expression changes were observed in the infected tumor cells. The infected 2BS cells showed no such changes. The colony formation ability in vitro was also reduced.]Conclusion: The reduced expression of cyclin D1 gene could induce human lung adenocarcinoma cells specific G1 arrest and apoptosis, while has no effect on normal cell.
1999, 6(2):97-101.
DOI: 10.3872/j.issn.1007-385X.1999.2.005
Abstract:
In primary hepatoma, the transcription of transthyretin (TTR) is seriously suppressed, and TTR decreased obviously in patients′ serum. It was necessary for us to purify TTR from human serum and observe the inhibitory effect of TTR on the growth of human hepatoma cells. Methods: The serum was precipitated in 40% saturated ammonium sulfate and 5% phenol, and the top layer was added to DEAE-Cellulose, then the protein was obtained. It was examined by Western blot. The inhibitory effect of TTR on the growth of human hepatoma cells was observed. Results: TTR was verified by Western blot and the purity of the TTR was at least 85%. It was confirmed that TTR inhibited evidently the growth of SMMC-7721 hepatoma cell. Conclusion: The evidently inhititory effect of TTR on SMMC-7721 hepatoma cell provided evidence for the feasibility of further genetic engineering production of TTR in future clinical therapy application. Italso supposed that TTR gene could be the candidate of the anti-oncogene related to human hepatoma
In primary hepatoma, the transcription of transthyretin (TTR) is seriously suppressed, and TTR decreased obviously in patients′ serum. It was necessary for us to purify TTR from human serum and observe the inhibitory effect of TTR on the growth of human hepatoma cells. Methods: The serum was precipitated in 40% saturated ammonium sulfate and 5% phenol, and the top layer was added to DEAE-Cellulose, then the protein was obtained. It was examined by Western blot. The inhibitory effect of TTR on the growth of human hepatoma cells was observed. Results: TTR was verified by Western blot and the purity of the TTR was at least 85%. It was confirmed that TTR inhibited evidently the growth of SMMC-7721 hepatoma cell. Conclusion: The evidently inhititory effect of TTR on SMMC-7721 hepatoma cell provided evidence for the feasibility of further genetic engineering production of TTR in future clinical therapy application. Italso supposed that TTR gene could be the candidate of the anti-oncogene related to human hepatoma
1999, 6(2):102-106.
DOI: 10.3872/j.issn.1007-385X.1999.2.006
Abstract:
To study the induced condition and characteristics of T cell anergy in vitro. Methods: Anergic Tcell was induced by combination of B7-1 mAb and cyclosporin A (CsA) in vitro, cytokine gene of anergic T cells was detected by RT-PCR. Results: T cell anergy was antigen-specific. The state of T cell anergy can be reversed by PHA, CD3 mAb and PMA plus A23187. IL-2 can prevent the induction of T cell anergy, but it can not reverse the state of un-responsiveness. IL-2 and IFN mRNA can not express in anergic T cells. In contrast, IL-4 and IL-10 mRNA were detectable. Conclusion: T cell anergy can be induced in vitro , cytokine profile of anergic T cells deviated to Th2-like phe-norype.
To study the induced condition and characteristics of T cell anergy in vitro. Methods: Anergic Tcell was induced by combination of B7-1 mAb and cyclosporin A (CsA) in vitro, cytokine gene of anergic T cells was detected by RT-PCR. Results: T cell anergy was antigen-specific. The state of T cell anergy can be reversed by PHA, CD3 mAb and PMA plus A23187. IL-2 can prevent the induction of T cell anergy, but it can not reverse the state of un-responsiveness. IL-2 and IFN mRNA can not express in anergic T cells. In contrast, IL-4 and IL-10 mRNA were detectable. Conclusion: T cell anergy can be induced in vitro , cytokine profile of anergic T cells deviated to Th2-like phe-norype.
1999, 6(2):107-110.
DOI: 10.3872/j.issn.1007-385X.1999.2.007
Abstract:
To construct eukaryotic expression vector of tumor-associated gene SNC90 and transfect it into human colorectal cancer cell lines. Methods: A 1.5 kb tumor-associate gene SNC90 full length cDNA was inserted into a mammalian expression vector pREP9 to make recombinant vector pREP9-SNC90, which was then introduced into three kinds of colorectal cancer cell lines, SW1116, COLO205 and SW620, by lipofection or electroporation. The cells resistant to G418 drug were selected. Results: Cells transfected with pREP9-SNC90 showed fewer G418-resistant colonies than those transfected with void vector, the inhibitory rates are 72.2%, 74.2% and 59.7%, respectively. Conclusion: SNC90 may play a negative role in regulating growth of colorectal cancer cell.
To construct eukaryotic expression vector of tumor-associated gene SNC90 and transfect it into human colorectal cancer cell lines. Methods: A 1.5 kb tumor-associate gene SNC90 full length cDNA was inserted into a mammalian expression vector pREP9 to make recombinant vector pREP9-SNC90, which was then introduced into three kinds of colorectal cancer cell lines, SW1116, COLO205 and SW620, by lipofection or electroporation. The cells resistant to G418 drug were selected. Results: Cells transfected with pREP9-SNC90 showed fewer G418-resistant colonies than those transfected with void vector, the inhibitory rates are 72.2%, 74.2% and 59.7%, respectively. Conclusion: SNC90 may play a negative role in regulating growth of colorectal cancer cell.
Chen Jiquan
,
Cao Xuetao
,
Xiu Qingyu
,
He Long
,
Zhang Weiping
,
Yu Yizhi
,
Zhang Minghui
,
Gu Shen
,
Luo Wentong
1999, 6(2):111-116.
DOI: 10.3872/j.issn.1007-385X.1999.2.008
Abstract:
To investigate the phenotypes and immune stimulating activity of IL-18 gene modified dendritic cells(DC). Methods: The expression of IL-18 mRNA was detected by RT-PCR in bone marrow DC from C57BL/6 mice after transfection with advenovirus vector containing IL-18 gene, and the IL-18 in their culture supernatant was determined by ELISA, also the induction of IFN-γ production by T cells with the culture supernatant of IL-18 gene modified DC was determined. The surface molecular expression and allogeneic MLR of IL-18 gene modified DC were investigated by FACS and ［3H］-TdR incorporation method respectively. Results： IL-18 gene modified DC could secret IL-18 effectively, IL-18 mRNA could be detected in IL-18 gene modified DC, and the IL-18 gene modified DC could induce T cells to produce IFN-γ. The allogeneic MLR activity of DC increased significantly after transfection with IL-18 gene, and the activity could only be partly blocked by IL-18 antibody. FACS analysis showed that some surface molecules such as Ia，B7.2，ICAM-1，CD40 of DC were upregulated significantly after transfection with IL-18 gene. Conclusion： IL-18 gene modified DC could secret IL-18 effectively with normal function such as induction of IFN-γ production from T cells. Some surface molecules on DC were upregulated and allogeneic MLR activity increased after transfection with IL-18 gene. The data suggested that IL-18 gene-modified DC might have enhanced antigen presenting capacity.
To investigate the phenotypes and immune stimulating activity of IL-18 gene modified dendritic cells(DC). Methods: The expression of IL-18 mRNA was detected by RT-PCR in bone marrow DC from C57BL/6 mice after transfection with advenovirus vector containing IL-18 gene, and the IL-18 in their culture supernatant was determined by ELISA, also the induction of IFN-γ production by T cells with the culture supernatant of IL-18 gene modified DC was determined. The surface molecular expression and allogeneic MLR of IL-18 gene modified DC were investigated by FACS and ［3H］-TdR incorporation method respectively. Results： IL-18 gene modified DC could secret IL-18 effectively, IL-18 mRNA could be detected in IL-18 gene modified DC, and the IL-18 gene modified DC could induce T cells to produce IFN-γ. The allogeneic MLR activity of DC increased significantly after transfection with IL-18 gene, and the activity could only be partly blocked by IL-18 antibody. FACS analysis showed that some surface molecules such as Ia，B7.2，ICAM-1，CD40 of DC were upregulated significantly after transfection with IL-18 gene. Conclusion： IL-18 gene modified DC could secret IL-18 effectively with normal function such as induction of IFN-γ production from T cells. Some surface molecules on DC were upregulated and allogeneic MLR activity increased after transfection with IL-18 gene. The data suggested that IL-18 gene-modified DC might have enhanced antigen presenting capacity.
1999, 6(2):117-120.
DOI: 10.3872/j.issn.1007-385X.1999.2.009
Abstract:
AbstractObjective: Many gene therapy protocols can induce antitumor immunity, however, the ex vivo approach restricts their uses. This sutydy intended to induce antitumor immunity by direct transfer of MHC class Ⅱ gene in vivo. Methods: MHC class Ⅱ gene cDNA was introduced directly into two tumors: P815, (a murine weak immunogenic mastocytoma) and B16 (a murine nonimmunogenic melanoma) to observe the survival rate of the mice. Results: Tumorigenicity of P815 was reduced when MHC class Ⅱ gene was introduced directly into tumors in vivo. Further more, most vaccinated mice could survive after second challenge of P815. Co-injection of MHC class Ⅱ and B7 genes in the B16 also resulted in the tumor grow slowly, while the injection of MHC class Ⅱ gene was not enough to induce effective antitumor responses. Conclusion: The results showed the potential applications of direct transfer of MHC class Ⅱ gene in the treatment of tumor.
AbstractObjective: Many gene therapy protocols can induce antitumor immunity, however, the ex vivo approach restricts their uses. This sutydy intended to induce antitumor immunity by direct transfer of MHC class Ⅱ gene in vivo. Methods: MHC class Ⅱ gene cDNA was introduced directly into two tumors: P815, (a murine weak immunogenic mastocytoma) and B16 (a murine nonimmunogenic melanoma) to observe the survival rate of the mice. Results: Tumorigenicity of P815 was reduced when MHC class Ⅱ gene was introduced directly into tumors in vivo. Further more, most vaccinated mice could survive after second challenge of P815. Co-injection of MHC class Ⅱ and B7 genes in the B16 also resulted in the tumor grow slowly, while the injection of MHC class Ⅱ gene was not enough to induce effective antitumor responses. Conclusion: The results showed the potential applications of direct transfer of MHC class Ⅱ gene in the treatment of tumor.
1999, 6(2):121-124.
DOI: 10.3872/j.issn.1007-385X.1999.2.010
Abstract:
AbstractObjective: To investigate the long-term protection of the bone marrow cells containing mutant dhfr gene on the reconstruction of murine hemacytogenesic function. Methods: The surviving mice with the tranduced marrow containing mutant hdhfr gene were used as donors for the third generation transplantation. The third transplant was carried out 14 weeks after secondary BMT(bone marrow transplantation). After irradiation(0.85～0.9 Gy) and BMT, the tertiary recipients, C57BL/6J, were treated with methotrexate (4 mg?kg -1 twice a week for the first week; 10 mg?kg -1 twice a week for the other weeks). The survival rate and blood pictures of murine as well as the integration and expression of target gene were studied. Results: The lethal irradiated murine hemacytogenesic function could be reinstituted and protected from lethal toxicity of high doses methotrexate selection after reinfusing the hematopoietic progenitor cells containing the hdhfr gene. The survival rate and survival time of experimental murine was higher than the control group. Survival rate of tertiary generation murine(40%) was less than the primary and secondary generations(87.5%). It was confirmed that hdhfr gene was still integrated into murine genomic DNA and expressed successfully using PCR and Southern blot analysis. Conclusion: he stable integration of human mutant dhfr gene may play an important role in this long term protection of murine hematopoietic function.
AbstractObjective: To investigate the long-term protection of the bone marrow cells containing mutant dhfr gene on the reconstruction of murine hemacytogenesic function. Methods: The surviving mice with the tranduced marrow containing mutant hdhfr gene were used as donors for the third generation transplantation. The third transplant was carried out 14 weeks after secondary BMT(bone marrow transplantation). After irradiation(0.85～0.9 Gy) and BMT, the tertiary recipients, C57BL/6J, were treated with methotrexate (4 mg?kg -1 twice a week for the first week; 10 mg?kg -1 twice a week for the other weeks). The survival rate and blood pictures of murine as well as the integration and expression of target gene were studied. Results: The lethal irradiated murine hemacytogenesic function could be reinstituted and protected from lethal toxicity of high doses methotrexate selection after reinfusing the hematopoietic progenitor cells containing the hdhfr gene. The survival rate and survival time of experimental murine was higher than the control group. Survival rate of tertiary generation murine(40%) was less than the primary and secondary generations(87.5%). It was confirmed that hdhfr gene was still integrated into murine genomic DNA and expressed successfully using PCR and Southern blot analysis. Conclusion: he stable integration of human mutant dhfr gene may play an important role in this long term protection of murine hematopoietic function.
Wu Chaowei
,
Zhang Yi
,
Huang Hualiang
,
Zhang Mingsheng
,
Huang Haomin
,
Lin Qing
,
Yu Xiaocong
,
Chen Ai
,
Li Xinyuan
,
Wang Dengshun
1999, 6(2):125-130.
DOI: 10.3872/j.issn.1007-385X.1999.2.011
Abstract:
Abstract Objectives: To construct a single-chain antibody (scFv) against lung adenocarcinoma and research its expressing conditions for establishing a clinical base. Methods: The variable genes of McAb anti-human lung adenocarcinoma, which were cloned from hybridoma cell WLA-2C4, were inserted into phagemid pFUW80 and expressed scFv in XL1-Blue. Also the scFv genes were inserted into another two vectors pTH and pTF. The corresponding host strains were Top10 and GI724/GI698, the inducing agents were IPTG and tryptophan, respectively. SDS-PAGE and ELISA methods were used to assay the expression products-phage antibodies in XL1-Blue and inclusion bodies in Top10 and GI724/GI698. Results: The scFv had the same specificity of the original monoclonal antibody against the antigen of A549 cell. The inclusion bodies were obtained at high yield 18.6% of the total E.coli proteins. Conclusion: A scFv against lung adenocarcinoma was successfully constructed and expressed in E. coli.
Abstract Objectives: To construct a single-chain antibody (scFv) against lung adenocarcinoma and research its expressing conditions for establishing a clinical base. Methods: The variable genes of McAb anti-human lung adenocarcinoma, which were cloned from hybridoma cell WLA-2C4, were inserted into phagemid pFUW80 and expressed scFv in XL1-Blue. Also the scFv genes were inserted into another two vectors pTH and pTF. The corresponding host strains were Top10 and GI724/GI698, the inducing agents were IPTG and tryptophan, respectively. SDS-PAGE and ELISA methods were used to assay the expression products-phage antibodies in XL1-Blue and inclusion bodies in Top10 and GI724/GI698. Results: The scFv had the same specificity of the original monoclonal antibody against the antigen of A549 cell. The inclusion bodies were obtained at high yield 18.6% of the total E.coli proteins. Conclusion: A scFv against lung adenocarcinoma was successfully constructed and expressed in E. coli.
1999, 6(2):131-135.
DOI: 10.3872/j.issn.1007-385X.1999.2.012
Abstract:
Abstract Objective: To examine if human acute promyelocytic leukemia cells (HL-60) apoptosis of could be induced by icarrin(ICA, a mononer purified from Epimedium Koreanum Nakai). Methods: The cells undergoing apoptosis were determined by morphology, DNA gel electrophoresis and flow cytometry (FCM). The expression of apoptosis associated genes bcl-2 and c-myc were evaluated by FCM and semi-quantitative RT-PCR methods. Results: ICA induced apoptosis showed classical morphological and biochemistry features in human cancer cells in a time and dose dependent manner in vitro. While inducing apoptosis, ICA downregulated expression of apoptosis associated genes bcl-2 and c-myc. Conclusion: This study suggests that ICA induced apoptosis of HL-60 cells is associated with downregulation of bcl-2and c-myc genes mRNA and protein expression.
Abstract Objective: To examine if human acute promyelocytic leukemia cells (HL-60) apoptosis of could be induced by icarrin(ICA, a mononer purified from Epimedium Koreanum Nakai). Methods: The cells undergoing apoptosis were determined by morphology, DNA gel electrophoresis and flow cytometry (FCM). The expression of apoptosis associated genes bcl-2 and c-myc were evaluated by FCM and semi-quantitative RT-PCR methods. Results: ICA induced apoptosis showed classical morphological and biochemistry features in human cancer cells in a time and dose dependent manner in vitro. While inducing apoptosis, ICA downregulated expression of apoptosis associated genes bcl-2 and c-myc. Conclusion: This study suggests that ICA induced apoptosis of HL-60 cells is associated with downregulation of bcl-2and c-myc genes mRNA and protein expression.
1999, 6(2):136-140.
DOI: 10.3872/j.issn.1007-385X.1999.2.013
Abstract:
Abstract Objective: To investigate effects of rhIL-17 on growth and development of mouse bone marrow progenitors and human cord blood CD34+ stem cells. Methods: Mouse bone marrow progenitors were isolated by routine protocol, and CD34+ stem cells were isolated from normal human cord blood by Mini-MACS， then cultured with rhIL-17 and/or GM-CSF/IL-4. The phenotypes of the cells were analyzed by FACS，IL-12 level was analyzed by ELISA and T cell stimulating activity in allo-MLR was determined by ［3H］-TdR incorporation. Results: Expression of MHC class Ⅱ molecules and B7-2 on the surface of immature DC derived from mouse bone marrow progenitors was up-regulated by IL-17. The capacity of the cells to secrete IL-12 and their T cell stimulating activity were also enhanced. The cells showed the characteristics of mature DC. After cultured with IL-17 for 9 days, the number of CD34+ stem cells increased by 2 times. The phenotypes of some cells were CD1ahigh, B7-2high,and HLA-DRlow. The cells could stimulate allo geneic T cells to proliferate but their capacity was lower than that of the cells cultured with IL-17 combined with GM-CSF. The cells cultured with IL-17 and GM-CSF proliferated markedly and the rate of CD1a+ and B7-2+ cells increased significantly. The T cell stimulating activity of cells was also augmented. Conclusion: IL-17 could promote DC derived from mouse bone marrow progenitors to mature. When combined with GM-CSF,IL-17 could induce human CD34+ stem cells not only to proliferate markedly but also to show characteristics of DC, indicating that CD34+ stem cells might differentiate to DC by IL-17.
Abstract Objective: To investigate effects of rhIL-17 on growth and development of mouse bone marrow progenitors and human cord blood CD34+ stem cells. Methods: Mouse bone marrow progenitors were isolated by routine protocol, and CD34+ stem cells were isolated from normal human cord blood by Mini-MACS， then cultured with rhIL-17 and/or GM-CSF/IL-4. The phenotypes of the cells were analyzed by FACS，IL-12 level was analyzed by ELISA and T cell stimulating activity in allo-MLR was determined by ［3H］-TdR incorporation. Results: Expression of MHC class Ⅱ molecules and B7-2 on the surface of immature DC derived from mouse bone marrow progenitors was up-regulated by IL-17. The capacity of the cells to secrete IL-12 and their T cell stimulating activity were also enhanced. The cells showed the characteristics of mature DC. After cultured with IL-17 for 9 days, the number of CD34+ stem cells increased by 2 times. The phenotypes of some cells were CD1ahigh, B7-2high,and HLA-DRlow. The cells could stimulate allo geneic T cells to proliferate but their capacity was lower than that of the cells cultured with IL-17 combined with GM-CSF. The cells cultured with IL-17 and GM-CSF proliferated markedly and the rate of CD1a+ and B7-2+ cells increased significantly. The T cell stimulating activity of cells was also augmented. Conclusion: IL-17 could promote DC derived from mouse bone marrow progenitors to mature. When combined with GM-CSF,IL-17 could induce human CD34+ stem cells not only to proliferate markedly but also to show characteristics of DC, indicating that CD34+ stem cells might differentiate to DC by IL-17.
1999, 6(2):141-144.
DOI: 10.3872/j.issn.1007-385X.1999.2.014
Abstract:
Abstract Objective: Through the improvement on determination of rhTNF-α bioassay, we established the formal procedure. Methods: We choose L929 cell for rhTNF-α bioassay determination. To identify the influence of rhTNF-α on the growth of HL-60 cells, various concentration and effecting time were studied. Results: The results showed that the inhibition of rhTNF-α on HL-60 cells based on dose and time. Northern Dot Blot and DNA electrophoresis showed c-myc gene was surpressed, and DNA was damaged by rhTNF-α. Conclusion: Autometic calculation is responsible for bioassay of rhTNF-α. rhTNF-α could inhibit the proliferation of HL-60 cells and its mechanism is different from doxirubicin.
Abstract Objective: Through the improvement on determination of rhTNF-α bioassay, we established the formal procedure. Methods: We choose L929 cell for rhTNF-α bioassay determination. To identify the influence of rhTNF-α on the growth of HL-60 cells, various concentration and effecting time were studied. Results: The results showed that the inhibition of rhTNF-α on HL-60 cells based on dose and time. Northern Dot Blot and DNA electrophoresis showed c-myc gene was surpressed, and DNA was damaged by rhTNF-α. Conclusion: Autometic calculation is responsible for bioassay of rhTNF-α. rhTNF-α could inhibit the proliferation of HL-60 cells and its mechanism is different from doxirubicin.
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.