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Cun-liang Deng

Publications and source records attributed to Cun-liang Deng.

6 recordsLinked to original sources
PubMed2006

[Construction, expression and immunogenicity study of a chimeric MS/hIL-12 eukaryotic expression plasmid].

OBJECTIVE: To construct and express a chimeric Mtb8.4 with signal peptide (MS)/hIL12 eukaryotic expression plasmid, and to study the immunogenicity of the MS/hIL-12 chimeric genetic vaccines. METHODS: The MS/hIL-12 chimeric gene was amplified by polymerase chain reaction (PCR) and cloned into the eukaryotic expression vector pCI-neo. The correct pCI-neo-MS/hIL12 (pMSI) recombinant plasmid was identified by PCR, restricted enzyme digestion and DNA sequencing. COS-7 cells were transfected with pMSI constructs by cationic liposome. After 48 hours, mRNA of the target gene was detected by RT-PCR, and hIL-12 protein in culture supernatant and cell lysates was detected by Western blot. C57BL/6N mice were vaccinated with MS/hIL-12 chimeric gene vaccine for three times at 3 week intervals. Four weeks after the final inoculation, three mice were sacrificed for measurement of the cytokine response and cytotoxic T lymphocyte (CTL) induction. RESULTS: The accuracy of plasmid construction was confirmed by a number of molecular biological techniques. Transfection of COS-7 cells with plasmids pMSI lead to transient expression of fusion proteins. The IFN-gamma and IL-2 titers were (1,521 +/- 48) ng/L and (755 +/- 41) ng/L in MS/hIL-12 chimeric gene vaccine group, (820 +/- 50) ng/L and (297 +/- 31) ng/L in MS gene vaccine group, (1,487 +/- 40) ng/L and (767 +/- 50) ng/L in BCG group, (121 +/- 16) ng/L and (62 +/- 10) ng/L in vacant vector group, and (48 +/- 16) ng/L and (32 +/- 17) ng/L in PBS group respectively. The levels of IFN-gamma and IL-2 in MS/hIL-12 chimeric gene vaccine group were higher than those of MS gene vaccine group, vacant vector group and PBS group (P < 0.01) and was similar to the BCG group (P > 0.05). The level of IL-4 in BCG group [(91 +/- 11) ng/L] increased significantly as compared to other groups (P < 0.01). When effector-cell-to-target-cell ratio (E:T ratio) were 100:1, 50:1, and 10:1 respectively, the CTL activity was 77.5%, 51.2%, 30.3% in MS/hIL-12 chimeric gene vaccine group, 56.2%, 37.8%, 11.5% in MS gene vaccine group, 28.9%, 21.4%, 9.8% in BCG group. The cytotoxicity in MS/hIL-12 chimeric gene vaccine group was higher than that of other groups (P < 0.01). CONCLUSION: When used to construct the chimeric gene vaccine, hIL-12 could improve the immunogenicity of MS gene vaccine.

Animals↗
PubMed2006

[Immunogenicity of chimeric gene vaccine Mtb8.4/hIL12].

AIM: To construct chimeric gene vaccine Mtb8.4/hIL-12, express it in COS-7 cells and study its immunogenicity. METHODS: Chimeric gene Mtb8.4/hIL-12 was amplified by PCR and cloned into the eukaryotic vector pCI-neo to construct the recombinant plasmid pCI-neo-Mtb8.4/hIL12. After the recombinant plasmid was identified by restriction enzyme digestion analysis, PCR and DNA sequencing, COS-7 cells were transfected with pCI-neo-Mtb8.4/hIL12 through cationic liposome. 48 hours later, the expression of mRNA was detected by RT-PCR and the level of hIL-12 in culture supernatant and cell lysates were detected by Western blot. C57BL/6N mice were vaccinated with chimeric gene vaccine Mtb8.4/hIL-12 three times at the interval of 3 weeks each time. Four weeks after the final inoculation, three mice were sacrificed to assess the cytotoxicity of CTLs and response to cytokine. RESULTS: The recombinant plasmid pCI-neo-Mtb8.4/hIL12 was constructed successfully. After COS-7 cells were transfected with pCI-neo-Mtb8.4/hIL12, chimeric gene Mtb8.4/hIL12 was expressed in COS-7 cells. The chimeric gene vaccine could induce strong antigen-specific immune response. With the increase of IFN-gamma and IL-2 secretion and the decrease of IL-4 secretion, the cytotoxicity of specific CTLs was heightened. CONCLUSION: Recombinant plasmid pCI-neo-Mtb8.4/hIL12 has been successfully constructed and expressed in COS-7 cells. The constructed chimeric gene vaccine Mtb8.4/hIL12 is of strong immunogenicity and can obviously induce the cytotoxicity of CTLs.

Animals↗
PubMed2005

[The relationship between TNF-alpha and the injury of renal tubular cells caused by anoxia/reoxygenation].

AIM: To investigate the relationship between TNF-alpha and renal tubular cell injury caused by anoxia/reoxygenation. METHODS: Human renal proximal tubular cell line HK-2 was used as model. Anoxia/reoxygenation were produced by covering/de-covering the cell culture with liquid paraffin wax. The level of TNF-alpha and the activity of lactate dehydrogenase(LDH) in the culture medium was determined by radioimmunoassay(RIA) and biochemical methods, respectively. Trypan blue exclusion was used to measure cell viability. RESULTS: Anoxia/reoxygenation could increase TNF-alpha level and LDH activity, but decrease viability of HK-2 cells. TNF-alpha level was positively correlated with LDH activity (r=0.89, P<0.05) and negatively with the cell viability (r=-0.91, P<0.05). CONCLUSION: TNF-alpha induced by anoxia/reoxygenation may participate in the process of renal tubular cell injury.

Cell Hypoxia↗
PubMed2005

[Role of interleukin-1 receptor antagonist in protecting kidney from injury induced by asphyxia in neonatal rats].

OBJECTIVE: To observe the role of recombinant human interleukin-1 receptor antagonist (rhIL-1ra) in protecting kidney from injury induced by asphyxia in neonatal rats. METHODS: Neonatal rats were used as experimental animals. The changes in intrarenal inflammatory response and renal injury were examined in the control group (n=13), and 2, 24 and 48 hours after asphyxia followed by normal saline treatment in those treated with rhIL-1ra. RESULTS: In normal saline group, the white blood cell count, the blood interleukin-1 (IL-1), IL-8, IL-6, nitric oxide (NO), endothelin (ET-1) levels, and the renal coefficient (LRC), the scores of injured tubules of the left kidney were significantly increased at 2 hours (n=10), 24 hours (n=11), and 48 hours (n=10, P<0.05 or P<0.01). Compared with the normal saline group, the levels of the above parameters, except IL-6, were significantly decreased in rhIL-1ra treatment group at the same time points (P<0.05 or P<0.01). Serum IL-6 at 24 hours and 48 hours was also decreased in rhIL-1ra treatment group significantly (P<0.05 or P<0.01). CONCLUSION: The results suggest that rhIL-1ra may protect renal injury after asphyxia via inhibiting intrarenal inflammatory response.

Animals↗