Biomedical subjects

Samuel Chak-Sum Cheng

Publications and source records attributed to Samuel Chak-Sum Cheng.

4 recordsLinked to original sources

PubMed2006

A DNA vaccine constructed with human papillomavirus type 16 (HPV16) E7 and E6 genes induced specific immune responses.

OBJECTIVE: Cervical cancer is found highly associated with human papillomaviruses type 16 (HPV16). HPV16 E6 and E7 oncogenes are important transforming genes which have become the main focus of anti-cervical cancer therapy. In this study, a recombinant DNA vaccine candidate, termed HPV16-DNA-E6E7, constructed with HPV16 E7 and E6 genes was generated and used to against HPV16-induced tumors. METHODS: We inserted an E7 DNA fragment into E6 gene to produce a recombinant gene (E6E7-DNA). The E6E7-DNA gene was inserted into a mammalian expression vector, pcDNA 3.1+, to construct the DNA vaccine candidate. Animals (C57BL/6 mice) were immunized with the vaccine candidate with various concentrations (50 microg, 100 microg or 200 microg, respectively), and cytotoxicity measurement and tumor protection assay were carried out to examine the immunological effects of the vaccine candidate. RESULTS: Immunization of with HPV16-E6E7-DNA induced HPV16-specific immune response and also conveyed protection against TC-1 induced tumor in vivo. A survival rate (90%) after 45 days of tumor challenge was observed. The animals injected with a higher dosage of the vaccine (200 microg) exhibited prolonged survival duration of more than 55 days. No transforming activity of the vaccine candidate was detected, as determined by focus formation and degradation of endogenous p53. CONCLUSION: Our results demonstrated that the HPV16-E6E7-DNA compound might become a candidate for HPV16 precautionary and immunotherapy.

Animals↗

PubMed2005

P-glycoprotein expression induced by glucose depletion enhanced the chemosensitivity in human hepatocellular carcinoma cell-lines.

Chemoresistance in cancer cells is frequently associated with an over-expression of the P-glycoprotein (P-gp). The expression of P-gp can be regulated as the cells encounter a number of chemical, physical or environmental stimuli. In this study, P-gp was found gradually expressed in a human hepatocellular carcinoma (HCC) QGY-7703 cells after 48 h of culturing in glucose-free medium. This phenomenon disappeared after the removal of glucose deprivation culture conditions. Mdr1-cDNA isolated from the cell line cultured in glucose-free conditions (namely QGY-7703G), was transiently transformed into the parent QGY-7703 cells, and multi-drug resistance was eventually induced. Results from XTT cytotoxicity assays indicated that the mdr1 gene was functional and the P-gp could restore the QGY-7703 cell's ability to withstand high concentrations of a number of chemotherapeutic agents. A P-gp inhibitor, verapamil, could completely reverse the cellular drug resistance when applied to the QGY-7703G cells. Our results indicated that an alteration of a specific state in cells caused by an external stimulus in vitro may lead to an expression of stress proteins (e.g. P-gp), which may enhance the cells' survival in adverse conditions. The expressed P-gp induced by glucose deprivation has a functional role in affecting the chemosensitivity in HCC QGY-7703G cells. Inhibition of P-gp activity may enhance the effect of the cancer cells towards cancer chemotherapy.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2004

Plasmid encoding papillomavirus Type 16 (HPV16) DNA constructed with codon optimization improved the immunogenicity against HPV infection.

Human papillomavirus Type 16 (HPV16) infections can cause neoplasia, which is thought to be closely associated with the development of cervical cancers. In the study, we attempted to construct a DNA plasmid encoding a HPV16 capsid protein (L1) and a HPV16 oncoprotein (E7), which was capable of preventing HPV16 infection and eliminating HPV16-infected cells. A plasmid, L1E7hpSCA1, encoding the L1 and E7 genes with the codon usage optimized for mammalian cell expression, was constructed. Mutations were introduced into the E7 gene sequence for reducing its oncogenicity. C57BL/6 mice were intramuscularly immunized at tibialis anterior (TA) muscles with the newly constructed L1E7hpSCA1 plasmid. The immune responses induced by the L1E7hpSCA1 plasmid (with codon optimization) and a control L1E7pSCA1 plasmid (without codon optimization) were compared. It is shown that the L1E7hpSCA1 was able to induce much stronger immune responses than the L1E7pSCA1. Sera obtained from immunized animals were found to contain anti-HPV16 antibodies as detected by ELISA and hemagglutination inhibition (HAI) assays. Cytotoxicity and interferon-gamma assays showed that spleenocytes from immunized animals were able to recognize and lyze E7 expressing tumor TC-1 cells. Moreover, the growth of E7 expressing tumor mass was inhibited in vaccinated mice. In vivo tumor protection test indicated that tumor formation was prevented in the experimental animals (67%) after vaccination with L1E7hpSCA1, while for the control group injected with L1E7pSCA1 only and the animal group injected with pSCA1 only, tumor formation was observed in all experimental animals. Our results suggest that the L1E7h gene (with codon optimization) is more effective against HPV16 than the L1E7 gene (without codon optimization). The L1E7hpSCA1 plasmid was able to provide protection against E7 expressing tumor, and it might have the potential to be a vaccine candidate for HPV prevention.

Animals↗

PubMed2002

A DNA vaccine against foot-and-mouth disease elicits an immune response in swine which is enhanced by co-administration with interleukin-2.

A plasmid DNA vaccine candidate (pCEIS) encoding two foot-and-mouth disease virus (FMDV) VP1 epitopes (amino acid residues 141-160 and 200-213) has been demonstrated to have the ability to elicit both FMDV-specific T cell proliferation and neutralizing antibody against FMD in swine. In this study, the efficiency of the pCEIS DNA vaccine when administrated by intramuscularly injection in swine was confirmed, and the immunogenicity of the pCEIS vaccine candidate was found to be enhanced through co-administration with a newly constructed plasmid (pIL2S) encoding the swine interleukin-2 (IL-2) cDNA. The expression of the pIL2S plasmid was driven by a CMV promotor provided by a pcDNA3.1 vector. Swine IL-2 cDNA was cloned by RT-PCR from swine spleen cells. The pIL2S plasmid was expressed in COS-7 cells after 24 and 96h of transfection in vitro. In an animal trial, results from T cell proliferation assay indicated that the stimulation index (SI) in response to stimulation of FMDV proteins in the swine groups injected with pCEIS plus pIL2S (SI ranging from 9.9 to 15.5) were significantly higher than that with pCEIS alone (SI ranging from 3.3 to 6.6). However, there was no significant difference in FMDV-neutralizing antibody level detected in these two swine groups. Mouse protection tests (MPTs) showed that the blood sera from immunized swine injected with either pCEIS alone or pCEIS plus pIL2S were able to protect suckling mice from FMDV challenge, with protection levels ranging from 10(1) to 10(2) lethal dose 50 (LD(50)) M. In a direct FMDV challenge, all swines immunized with either pCEIS plus pIL2S or with pCEIS alone were challenged with 50LD(50)S (50 x lethal dosage in swine) of FMDV. The animals were fully protected (100%) from the FMD viral challenge. These results suggest that co-administration of the plasmids, pCEIS and pIL2S, enhances of the immunogenicity of the pCEIS DNA vaccine candidate, and both intramuscular injection of pCEIS alone and co-administration of the vaccine candidate with pIL2S can protect the swine from direct FMD challenge.

Animals↗