Influence on the immune function of the human peripheral blood mononuclear cells transfected by retrovirus-mediated HSV-tk gene.
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Cytokines↗
Biomedical subjects
Hang-ping Yao
Publications and source records attributed to Hang-ping Yao.
[Inhibitory effects of mizolastine on substance P-induced production of leukotriene B4 and interleukin 5 in mouse skin].
OBJECTIVE: To observe the inhibitory effect of mizolastine on substance P(SP)-induced production of leukotriene B(4) (LTB(4)) and interleukin 5 (IL-5) in mouse skin. METHODS: Mice were fed with different doses of mizolastine or other control drugs, 30 min after administration animals were injected intradermally with SP on the back. The treated skin samples were taken and competitive enzyme-link immunoassay (ELISA) method was applied to detect LTB (4) and IL-5 in the skin samples. RESULT: The LTB(4) and IL-5 levels in 10 mg/kg mizolastine group were (1.23 +/-0.29)pg/ml and (34.28 +/-11.00)pg/ml, respectively, which were lower than those in saline control group [(5.52+/-1.88)pg/ml and (179.62 +/-46.25)pg/ml respectively] or loratadine group [(3.89+/-1.27)pg/ml and (127.74 +/-43.27)pg/ml respectively]. No significant difference was found between 10 mg/kg mizolastine group and dexamethasone group (P=0.161 and P=0.508). CONCLUSION: Mizolastine might inhibit the production of LTB(4) and IL-5 induced by substance P in mouse skin, suggesting that anti-inflammatory effect and the blockade of histamine H1 receptors might be involved in its anti-pruritic mechanisms.
Animals↗
[Expression of VEGF165 and VEGF121 genes in MC3T3-E1 cell line and biological function of their products].
OBJECTIVE: To construct vectors stably expressing bioactive VEGF165 and VEGF121 in MC3T3-E1 cells. METHODS: VEGF165 and VEGF121 genes were obtained by RT-PCR from HL-60 cells and inserted into eukaryotic expression plasmid pcDNA3.1(+). The constructed plasmids containing right sequence of target genes were transfected into MC3T3-E1 cells by lipofectin. The cells were limitedly diluted twice and selected by G418, the expression of VEGF was tested by ELISA, Western-blot and immunofluorescence essay. The bioactivity of expressed products was proved by MTT method. RESULT: DNA sequencing indicated the sequences of the obtained VEGF165 and VEGF121 were identical to the those reported in Genebank, and the newly-constructed plasmids were VEGF165/pcDNA3.1(+) and VEGF121/pcDNA3.1(+). The results of ELISA, Western-blot and immunofluorescence showed that the transfected cells could express VEGF165 and VEGF121 proteins. And MTT proved the proteins were bioactive. CONCLUSION: The transfected MC3T3-E1 cells could stably express high levels of bioactive VEGF165 and VEGF121.
3T3 Cells↗
[Construction of expression vector of hTERT/hIL-18 fusion gene in eukaryotic cells and its function].
OBJECTIVE: To construct expression vector of hTERT-hIL-18 fusion gene in eukaryotic cells and to study its biological function. METHODS: hIL-18 gene was amplified by RT-PCR, then T-A cloned and inserted into PCDNA3.1(+)/hTERT vector. The sequence of fusion gene was examined by enzyme incision and DNA sequencing. The vector with fusion gene was transformed into 3T3 cells by the method of lipofecting, and proved by Western blot. The secretion gamma-interferon was measured with ELISA and cell apoptosis was detected with flow cytometry. RESULT: Expression vector PCDNA3.1(+) of hTERT/hIL-18 fusion gene was constructed successfully. The correct sequence was proved by enzyme incision and sequencing and there was a correct open reading frame. Fusion protein of hTERT/hIL-18 was effectively expressed in eukaryotic cells and was proved by Western blot and immunofluorescence stain. The fusion protein stimulated KG-1 cells to secrete gamma-interferon and had anti-apoptosis effect. CONCLUSION: Fusion protein hTERT-hIL-18 is highly effectively expressed in eukaryotic cells and is biologically active.
Base Sequence↗
[Construction and identification of recombinant adenovirus expressing rat proteoglycan II].
OBJECTIVE: To construct recombinant adenovirus expressing rat proteoglycan II (Ad-DCN) and to study its biological features. METHODS: Rat DCN gene was inserted into an E1 and E3-substituted adenovirus shuttle plasmid. And this shuttle plasmid including DCN was recombined with AdEasy-1 in BJ5183-AD-1 electroporation competent cells to form recombinant adenovirus-DCN plasmid, which was further transfected into Ad293 cells to passage adenovirus. The control recombinant adenovirus, Ad-LacZ, was also constructed in the same way. After plaque forming trail and adjusted with PBS, Ad-DCN and Ad-LacZ were obtained with titer 1x10(9) pfu x ml(-1). PCR, Western blot and MTT analysis were used to detect the expression of DCN or the bioactivity of expressed DCN. RESULT: DCN was detected in Ad-DCN infected CHO cells by PCR and Western blot, but not in Ad-LacZ infected CHO cells. MTT analysis results showed that the supernatant from the culture of Ad-DCN infected CHO cells could abrogate the inhibitive effect of TGFbeta1 on proliferation of CCL-64 cells. The proliferation rate of TGFbeta1 + Ad-DCN treated cells was significantly higher than that of TGFbeta1 + Ad-lacz or TGFbeta1 treated cells [(0.5252 +/-0.04 compared with 0.2826 +/-0.02 or 0.2918 +/-0.02) OD, P <0.05] and lower than that of control cells [(0.9332 +/-0.08) OD, P <0.05]. CONCLUSION: The constructed recombinant adenovirus can express biologically active decorin.
Adenoviridae↗
[Expression of recombinant plasmid of HIV-1 gp120 and IFN-gamma in E. coli and its immunoregulatory effect].
OBJECTIVE: To investigate the regulating effect of interferon-gamma (IFN-gamma) gene on the immune response induced by human immunodeficiency virus (HIV) gp120 in mice. METHODS: The recombinant prokaryotic plasmid pet44b with HIV-1gp120 N and pet44b with HIV-1gp120 N linking IFN-gamma were constructed and expressed in E. coli BL21 (DE3) through IPTG. The purified proteins gp120 N and gp120 N/IFN-gamma were used in the following experiments: mice were injected with PBS i. s as control group 1, injected i. s with gp120 N as control group 2, and injected i. s with gp120 N/IFN-gamma as experiment group. The serum samples and spleen cells were collected 2 weeks after completion of immune program.T cells enlargement stimulated by gp120, CTL test and the detection of Th1 cytokines (SKs) IL-2, IFN-gamma and Th2 CKs IL-4, IL-10 were performed. RESULT: SDS-PAGE and Western blot confirmed that gp120 N and gp120 N/IFN-gamma fusion proteins were successfully expressed. The immune experiments showed that the enlargement of specific T lymphocytes, the response of CTL and the expression of Th1 type CKs IL-2 and IL-gamma against gp120 were significantly different among the three groups (P < 0.05). And the response of gp120 N/IFN-gamma group was stronger than that of gp120 N group, the latter was stronger than that of PBS group (P < 0.05). The differences in expression of Th2 type CKs IL-4 and IL-10 were not significant among the three groups (P >0.05). CONCLUSION: The cell specific immune responses of BALB/c mice to HIV-1CNgp120 antigen can be enhanced when HIV-1CNgp120 gene and IFN-gamma gene are used in coordination.
Animals↗
[Comparison of immunological profiles between pediatric and adult patients with AIDS in China].
OBJECTIVE: To compare the immunological profiles of pediatric and adult patients with AIDS in China. METHODS: Totally 103 pediatric AIDS patients, 38 adult patients, 88 healthy children, and 72 healthy adults were enrolled. CD4 + T lymphocyte counts were determined by four-color flow cytometer and HIV-RNA levels were measured in EDTA plasma by quantitative reverse-transcription polymerase chain reaction (RT-PCR). Plasma levels of interleukin (IL)-10, IL-16, IL-18, regulated upon activation, normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein 1 (MCP-1), stromal cell-derived factor-(SDF-1) alpha, SDF-1 beta, and macrophage stimulate protein (MSP) were quantified by enzyme-linked immunosorbent assay (ELISA). The levels of beta 2-microglobulin (beta 2-MG) and soluble Fas (sFas) were measured to indicate the activation of immune system. RESULTS: The mean CD4 + T cell count in pediatric patients with AIDS was significantly lower than in healthy children (P < 0.01), as between the adult AIDS patients and healthy adults (P < 0.01). The mean levels of these cytokines in pediatric patients were significantly higher than in healthy children (P < 0.01). The level of MSP in adult patients was significantly lower than in healthy adults and other cytokines were significantly higher (P < 0.01). The mean levels of these cytokines, except SDF1 alpha and beta 2-MG, were significantly higher in pediatric patients than in adult patients (P < 0.01). CONCLUSION: Abnormal immune activation is induced in both pediatric and adult patients with HIV-1 infection. The level of immune activation is higher in pediatric patients than in adult patients.
Acquired Immunodeficiency Syndrome↗
Combination of small interfering RNAs mediates greater inhibition of human hepatitis B virus replication and antigen expression.
OBJECTIVES: To evaluate the inhibitory effect mediated by combination of small interfering RNAs (siRNAs) targeting different sites of hepatitis B virus (HBV) transcripts on the viral replication and antigen expression in vitro. METHODS: (1) Seven siRNAs targeting surface (S), polymerase (P) or precore (PreC) region of HBV genome were designed and chemically synthesized. (2) HBV-producing HepG2.2.15 cells were treated with or without siRNAs for 72 h. (3) HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay. (4) Intracellular viral DNA was quantified by real-time PCR (Polymerase Chain Reaction). (5) HBV viral mRNA was reverse transcribed and quantified by real-time PCR. (6) The change of cell cycle and apoptosis was determined by flow cytometry. RESULTS: Our data demonstrated that synthetic small interfering RNAs (siRNAs) targeting S and PreC gene could efficiently and specifically inhibit HBV replication and antigen expression. The expression of HBsAg and HBeAg and the replication of HBV could be specifically inhibited in a dose-dependent manner by siRNAs. Furthermore, our results showed that the combination of siRNAs targeting various regions could inhibit HBV replication and antigen expression in a more efficient way than the use of single siRNA at the same final concentration. No apoptotic change was observed in the cell after siRNA treatment. CONCLUSION: Our results demonstrated that siRNAs exerted robust and specific inhibition on HBV replication and antigen expression in a cell culture system and combination of siRNAs targeting different regions exhibited more potency.
Apoptosis↗
Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B.
OBJECTIVE: To construct a cDNA library from human liver tissue with chronic hepatitis B and check its quality for investigating the expression level of liver tissue infected by hepatitis B virus. This will then be used to find the relevant genes and interesting proteins associated with the development of hepatitis B. METHODS: The total RNA from liver tissue with chronic hepatitis B was extracted and the mRNA was purified using TRIZOL method. Switching mechanism at 5' end of the RNA transcript (SMART) technique and CDS III/3' primer were used for first-strand cDNA synthesis. Long distance polymerase chain reaction (LD PCR) was then used to synthesize the double-strand cDNA that was then digested by Sfi I and fractionated by CHROMA SPIN-400 column. The longer than 0.4 kb cDNAs were collected and ligated to lambdaTriplEx2 vector. Then lambda phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. Fourteen plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. RESULTS: The titers of unamplifed and amplified libraries were 1.94 x 10(6) pfu/ml and 1.49 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 98.15% in unamplified library and 98.76% in amplified library. The lengths of the inserts were 1.23 kb in average, 1-2 kb in 64.29%, and 0.5-1.0 kb in 35.71%. CONCLUSION: A high quality cDNA library from human liver tissue with chronic hepatitis B was successfully constructed.
DNA, Complementary↗
[Construction of the eukaryotic expression vector containing rat transforming growth factor beta type II receptor and interferon gamma fusion genes].
OBJECTIVE: To express a bioactive fusion protein comprising rat soluble transforming growth factor beta type II receptor and interferon gamma(rsTGFbetaR II-IFN gamma) in mammalian cells. METHODS: mRNA was extracted from rat liver and the sTGFbetaR II-IFN gamma genes amplified by RT-PCR, then the two gene segments were cloned into the same pSecTag2A expression vector, and pSecTag2A/rsTGFbetaR II-IFN gamma recombinant plasmid was obtained, which was later transfected into CHO cells using liposomes. The expression of pSecTag2A/rsTGFbetaR II-IFN gamma in the supernatant was detected by ELISA and Western blotting. The bioactivities of the fusion protein were tested by sTGF betaR II-IFN gamma and IFN gamma bioassays. RESULTS: pSecTag2A/rsTGFbetaR II-IFN gamma transfectants expressed rsTGF betaR II-IFN gamma fusion protein. The purified fusion protein exhibited anti-viral activity and antagonized the proliferation-inhibitive effect of TGFbeta1 on CCL-64 cells. It inhibits the HSC activation in vitro. CONCLUSION: The pSecTag2A/rsTGFbetaR II-IFN gamma recombinant plasmid constructed in this study can express bioactive rsTGFbetaR II-IFN gamma fusion protein.
Amino Acid Sequence↗
[Construction and characterization of a cDNA library from human liver tissue of cirrhosis].
OBJECTIVE: To construct a cDNA library from human liver tissue of cirrhosis. METHODS: The total RNA from human liver tissue of cirrhosis was extracted using Trizol method, and the mRNA was purified using mRNA purification kit. SMART technique and CDSIII/3' primer were used for first-strand cDNA synthesis. Long distance PCR was then used to synthesize the double-strand cDNA that was then digested by proteinase K and Sfi I, and was fractionated by CHOMA SPIN-400 column. The cDNA fragments longer than 0.4 kb were collected and ligated to lambdaTripl Ex2 vector. Then lambda-phage packaging reaction and library amplification were performed. The qualities of both unamplified and amplified cDNA libraries was strictly checked by conventional titer determination. Eleven plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. RESULTS: The titers of unamplifed and amplified libraries were 1.03 x 10(6) pfu/ml and 1.36 x 10(9) pfu/ml respectively. The percentages of recombinants from both libraries were 97.24 % in unamplified library and 99.02 % in amplified library. The lengths of the inserts were 1.02 kb in average (36.36 % 1 approximately equals 2 kb and 63.64 % 0.5 approximately equals 1.0 kb). CONCLUSION: A high quality cDNA library from human liver tissue of cirrhosis was constructed successfully, which can be used for screening and cloning new special genes associated with the occurrence of cirrhosis.
DNA, Complementary↗
[Inhibition of HBV core antigen gene expression in human embryonic kidney cell line AD293 by plasmid-based RNAi].
OBJECTIVE: To inhibit HBV core antigen gene expression with plasmid-based RNAi. METHODS: The shRNA expression vector targeting HBV core antigen gene was designed and constructed. Human embryonic kidney cell line AD293 was co-transfected with HBcAg-EGFP fusion protein expression vector and shRNA expression vector transiently, and the cells without shRNA-transfection and with non-specific shRNA transfection were used as controls. Inhibitory effect of RNAi was detected by fluorescence-activated cell sorting (FACS) and real-time fluorescence quantificational RT-PCR. RESULTS: HBV core antigen gene expression in AD293 was inhibited by shRNA, with the maximal inhibition rate of 76 % measured by FACS and of 63.1 % by real-time PCR. CONCLUSION: Effective inhibition of HBV core antigen gene expression by plasmid-based RNAi provides an alternative for anti-HBV study in vitro, which has potential clinical application.
Cell Line↗
Immunotherapy of intracranial G422 glioblastoma with dendritic cells pulsed with tumor extract or RNA.
OBJECTIVE: To investigate the anti-tumor efficacy of dendritic cell (DC)-based vaccines pulsed with tumor extracts or RNA in a mouse model of intracranial G422 glioblastoma. METHODS: Bone marrow-derived DCs were pulsed ex vivo with tumor extracts or RNA. Ninety female mice harboring 4-day-old intracranial G422 glioblastomas and 126 normal mice were treated with three spaced one week apart subcutaneous injections either with PBS, unpulsed DCs, G422 tumor extracts, RNA, DCs pulsed with G422 tumor extracts (DC/extract) or with RNA (DC/RNA). Seven days after the third immunization of normal mice, the spleens of 36 of them were harvested for cytotoxic T lyphocyte (CTL) assays and the others were challenged in the brain with G422 tumor cells. All the treated mice were followed for survival. Some mice brains were removed and examined pathologically when they died. RESULTS: Immunization using DC/extract or DC/RNA significantly induced G422-specific CTL responses compared with control groups (P<0.01). Vaccination with DC/extract or DC/RNA, either prior to G422 tumor challenge or in tumor-harboring mice, significantly prolonged survival compared with other control groups (P<0.01). CONCLUSION: DCs pulsed with tumor extracts or RNA derived from autologous tumors has potential antitumor effects via activation of cell-mediated immunity. Our results suggest a useful therapeutic strategy against gliomas.
Animals↗
[Effect of Triptolide on TNFalpha-induced activation of NF-kappaB and expression of COX-2 and iNOS in human rheumatoid arthritis synovial fibroblasts].
OBJECTIVE: To explore the effects of Triptolide (TP) on TNFalpha-induced cell proliferation and expressions of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and their inducing products PGE2, NO in human rheumatoid arthritis synovial fibroblasts (RASF). METHODS: Fibroblasts (RASF) were obtained from synovial tissue of patients with RA and were cultured in vitro. RASF were stimulated with TNFalpha(20 microg/L) in the presence or absence of TP(0 - 100 microg/L) for 20 h. The RASF proliferation was determined by (3)H-TdR incorporation, and the productions of PGE2 and NO in culture supernatants of RASF were detected with competitive ELISA and enzyme reduction of nitrate. Expressions of COX-2 and iNOS mRNA in RASF were analyzed by semi-quantitative RT-PCR. Expressions of COX-2 and iNOS protein were estimated by Western-blot method and cellular enzyme immunoassay in synovial fibroblasts. NF-kappaB activity in whole-cell extract of RASF was also measured by an ELISA-based method. RESULTS: TP (>20 microg/L) down-regulated markedly TNFalpha-induced COX-2 and iNOS mRNA and protein expression, and their inducing products PGE2 and NO of synovial fibroblasts. This effect was positively correlated with TP concentrations. NF-kappaB activity in TNFalpha-stimulated synovial cells was suppressed profoundly by TP treatment (IC(50) approximately 35microg/L). The activity of NF-kappaB was correlated with the levels of COX-2 and iNOS expression in TNFalpha-stimulated RASF. No change was observed in proliferation of synovial cells after treatment of TP. CONCLUSION: TP could significantly down-regulate TNFalpha-induced COX-2, iNOS expression and production of PGE2, NO in human RASF, which is associated with the suppression of NF-kappaB activity.
Arthritis, Rheumatoid↗
[Development of a system for quick screening of efficient HBx-siRNA].
OBJECTIVE: To develop a system for quick screening of efficient siRNA targeted HBx mRNA. METHODS: Using recombination DNA technique, the fusion expression plasmid of HBx and EGFP was constructed, and siRNA expression cassettes (SECs) containing U6+1, H1 or tRNA(Val )promoter were prepared via one-step overlapping extension PCR. By co-transfection with recombinant plasmid and SECs into AD293 cell, the inhibition effects on the transient expression of HBx-EGFP fusion protein were analyzed by FACS and semi-quantitated RT-PCR analysis. RESULT: (1)HBx-EGFP fusion protein expression plasmid pHBx-EGFP was constructed successfully, which expressed green fluorescence in cell mainly located at plasma or the periphery of nucleus in granules. (2) Co-transfection with recombinant plasmid and SECs into AD293 cells resulted in inhibition of HBx-EGFP expression. SEC-siHBx388 showed significant inhibition effect on HBx-EGFP expression compared with SEC-siHBx271, indicating that siHBx388 is effective siRNA site and could be screened out with our screening system. In addition,the results of that U6+1-, tRNA(Val) and H1-siHBx388 reduced HBx-EGFP expression by 21.7%, 12.9% and 12.4% of control respectively indicated that both tRNAVal and H1 promoter was high efficient in driving effect of siHBx388. CONCLUSION: Combination of the HBx expression carrying reporter gene and PCR-based multi promoter SECs may develop a useful system to be applied in identification of optimal HBx- siRNA and its matching promoter.
Base Sequence↗
[Construction and expression of the eukaryotic expression vector containing human soluble transforming growth factor beta receptor II].
OBJECTIVE: To construct a eukaryotic expression vector encoding the gene of extracellular region of type II transforming growth factor beta receptor (sTGFbetaR II), to express the protein in CHO cell line and to determine its biological activity. METHODS: The extracellular region (amino acids 1-159) of the human TGFbetaR II cDNA was amplified by PCR from a TGFbetaR II chimeric plasmid,and the eukaryotic expression plasmid pCDNA3.1/myc-his(-)B-sTGFbetaR II(pCDNA-sTGFbetaR II) was constructed by inserting the sTGFbetaR II cDNA into the EcoR I/Hind III-digested pCDNA. The DNA sequence was confirmed by double digestion and the pCDNA-sTGFbetaR II plasmid was transfected into the CHO cell line. The sTGFbetaR II protein was confirmed by Western blotting analysis and its biological function was determined. RESULTS: The specific protein was observed in western blotting, and the protein abrogated the growth-inhibitory effects of TGF-beta1 on mink lung epithelial cells (Mv1Lu). CONCLUSION: The eukaryotic expression plasmid pCDNA-sTGFbetaR II has been successfully constructed and the sTGFsTGFbetaR II protein with biological activity obtained.
Amino Acid Sequence↗
[Inhibitory effect of sinomenine on expression of cyclooxygenase-2 in lipopolysaccharide-induced PC-12 cells].
OBJECTIVE: To study the effects of sinomenine (Sin) on cell proliferation, intracellular expression of cyclooxygenase-2 (COX-2), and production of PGE2 in lipopolysaccharide-induced PC-12 cells, To explore the Sin's mechanism on nerve cell. METHOD: PC-12 cells were cultured with nerve growing factors (NGF), and pretreated with Sin at various concentrations (0, 3 x 10(-6), 30 x 10(-6), 150 x 10(-6) mol x L(-1)) for 2 hours, then with or without stimulation of lipopolysaccharide (LPS). The proliferation activity of PC-12 cells was determined by 3H-TdR incorporation, and the production of PGE2 in culture supernatants of PC-12 cells was detected with competitive ELISA. Expression of COX-2 mRNA in PC-12 cells was analyzed by semi-quantitative RT-PCR, and expression of COX-2 protein was estimated by Western blot method and cellular enzyme immunoassay. Nuclear factor-kappa B (NF-kappaB) activity in whole-cell extract of PC-12 cells was also measured by an ELISA-based method. RESULT: The data showed that Sin down-regulated the expression of COX-2 mRNA and protein, and reduced the production of PGE2 in the LPS-stimulated PC-12 cells which correlated with Sin's concentrations positively. In addition, NF-kappaB activity in LPS-stimulated cells was suppressed significantly by Sin. No inhibition of proliferation of PC-12 cells due to Sin treatment was observed. CONCLUSION: Sin mediates the down-regulation of expression of COX-2 and production of induced PGE2 in PC-12 cells by suppressing the activity of NF-kappaB.
Animals↗
[Subcutaneous transplantation of microencapsulated Chinese hamster ovary (CHO) cells/pcDNA 3.1/mIL-12 and subcutaneous transplantation of microencapsulated CHO/pcDNA3.1/mIL-12 combined with 5-fluoro-uracil in treatment of tumor-burdened mice].
OBJECTIVE: To investigate the effect of subcutaneous transplantation of microencapsulated Chinese hamster ovary cells (CHO)/pcDNA3.1/mIL-12 and subcutaneous transplantation of microencapsulated CHO/pcDNA3.1/mIL-12 combined with 5-fluoro-uracilum (5-FU) in treatment of tumor-burdened mice. METHODS: CHO/pcDNA3.1/mIL-12 and CHO/pcDNA3.1 were suspended in solution of sodium alginate and made into microcapsules. Sixty mice were divided into 6 groups of 10 mice: (1) IL-12 group, microencapsulated CHO/pcDNA3.1/mIL-12 was injected subcutaneously, (2) combined treatment group: CHO/pcDN with 5-FU, (3) 5-FU group: 5-FU was injected intraperitoneally, (4) blank vector group: CHO/pcDNA3.1 was injected subcutaneously, (5) tumor-burdened group: without any treatment, and (6) blank control group: normal mice without any treatment. Except the mice of the blank control group, all mice were injected subcutaneously into the inner side of right thigh with mice colonic adenoma cells. The volume of tumor was measured every third day. 20 days after the treatment, 5 mice in each group were killed to examine the serum Th1 type cytokines: interferon (IFN)-gamma, IL-12, and Th2 type cytokins: IL-4, IL-10 by double antibody sandwich ELISA. The spleens were made into suspension of lymphocytes to examine the activity of natural killer cell (NK) and cytolytic T lymphocyte (CTL). The survival period of the remaining 5 mice in each group was observed till the 60th day. RESULTS: The activity of NK and CTL were significantly much more in the IL-12 group than in other groups. The activity of NK was significantly much more in combined treatment group than in the tumor-burdened group. The levels of Th1 type cytokines were the lowest in the tumor-burdened group. There was no difference in the levels of Th1 type cytokine between the tumor-burdened group and 5-FU group. However, the levels of Th1 type cytokine were significantly higher in the IL-12 group than in other groups. The levels of Th2 type cytokines were rather high in the tumor-burdened group. There was no difference in the levels of Th2 type cytokine between the tumor-burdened group and 5-FU group. However, the levels of Th2 type cytokine were the lowest in the IL-12 group than in other groups. There was no significant difference in volume of tumor among the tumor-burdened group, blank vector group, and 5-FU group. The mean diameters of tumor in the IL-12 group and combined treatment group were significantly smaller than in the 5-FU, tumor-burdened, and blank vector groups (P < 0.05), however, without a difference between the IL-12 group and combined treatment group. The survival periods of the IL-12 group and combined treatment group were significantly longer than those in the blank vector, tumor-burdened and 5-FU groups (P < 0.05). CONCLUSION: Microencapsulated CHO/pcDNA3.1/mIL-12 transplanted subcutaneously significantly improves the immune function of tumor-burdened mice and partially overcomes immune suppression caused by chemotherapy, and is effective in slowing the growth of tumor and lengthening the survival period of tumor-burdened ice.
Animals↗