ABSTRACT
Objective To evaluate the efficacy of some medicinal plants and systemic glucantime in a comparative manner against the causative agent of cutaneous leishmaniasis both in vitro and in BALB/c mice. Methods For in vivo testing, inbred mice were challenged with Leishmania major parasites and the resultant ulcers were treated with extract based-ointments applied topically two times per day for a period of 20 days. A group of 56 mice were randomly divided into 7 subgroups. The control group received the ointment void of extracts, whereas the reference group received glucantime only. The efficacy of treatments was evaluated by measuring ulcer diameter, parasite burden and NO production. Results Our results indicated that plant extract based-ointments were effective in reducing ulcer size and parasite burden in spleens, but their effects did not differ significantly from that of glucantime. The plant extracts tested in this study were able to increase NO production that helped parasite suppression. Conclusions Our findings indicate that the tested plant extracts are effective against Leishmania major both during in vitro and in vivo experiments, but further researches are required to recommend a potential plant extract as an alternative drug.
ABSTRACT
BACKGROUND: TSA (thiol-specific antioxidant antigen) is the immune-dominant antigen of Leishmania major and is considered to be the most promising candidate molecule for a recombinant or DNA vaccine against leishmaniasis. The aim of the present work was to express a plasmid containing the TSA gene in eukaryotic cells. METHODS: Genomic DNA was extracted, and the TSA gene was amplified by polymerase chain reaction (PCR). The PCR product was cloned into the pTZ57R/T vector, followed by subcloning into the eukaryotic expression vector pcDNA3 (EcoRI and HindIII sites). The recombinant plasmid was characterised by restriction digest and PCR. Eukaryotic Chinese hamster ovary cells were transfected with the plasmid containing the TSA gene. Expression of the L. major TSA gene was confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting. RESULTS: The plasmid containing the TSA gene was successfully expressed, as demonstrated by a band of 22.1 kDa on Western blots. CONCLUSION: The plasmid containing the TSA gene can be expressed in a eukaryotic cell line. Thus, the recombinant plasmid may potentially be used as a DNA vaccine in animal models.
ABSTRACT
Background: TSA (thiol-specific antioxidant antigen) is the immune-dominant antigen of Leishmania major and is considered to be the most promising candidate molecule for a recombinant or DNA vaccine against leishmaniasis. The aim of the present work was to express a plasmid containing the TSA gene in eukaryotic cells. Methods: Genomic DNA was extracted, and the TSA gene was amplified by polymerase chain reaction (PCR). The PCR product was cloned into the pTZ57R/T vector, followed by subcloning into the eukaryotic expression vector pcDNA3 (EcoRI and HindIII sites). The recombinant plasmid was characterised by restriction digest and PCR. Eukaryotic Chinese hamster ovary cells were transfected with the plasmid containing the TSA gene. Expression of the L. major TSA gene was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and Western blotting. Results: The plasmid containing the TSA gene was successfully expressed, as demonstrated by a band of 22.1 kDa on Western blots. Conclusion: The plasmid containing the TSA gene can be expressed in a eukaryotic cell line. Thus, the recombinant plasmid may potentially be used as a DNA vaccine in animal models.
