Leishmania tarentolae Cell-Free System for Filamentous Protein Expression

Background: In earlier studies, Leishmania tarentolae is known as an expression system due to its post-translational modification. SAG1 and Cyc18 proteins of T. gondii was evaluated for protection against toxoplasmosis. A surface glycoprotein of T. gondii, is a promoter of the immune response and previously studied for vaccine development during the infection. Cyc18 induces IL-12, which can drive Th1 and CD8+ T cell development when bound to CCR5, it rolled to adjuvant the immune response of T. gondii surface protein. Objectives: This study was performed to evaluate the efficiency of SAG1 (Surface Antigen 1) and TgCyc18 (Cyclophilin 18) considered a a CCR5 chemochine for Toxoplasma gondii vaccine when expressed in L. tarentolae. Methods: SAG1 and TgCyc18 genes were cloned and transfected into L. tarentolae. Protein expression and secretion were effectively validated by PCR, immunoblotting, enzyme activity, ELISA, and immunofluor. And features of SAG1 were evaluated mass spectrometry. Results: Successful transfection of SAG1 and TgCyc18 plasmids and transfected into Leishmania tarentolae, and was confirmed by two steps of PCR. Current study verified that the culture supernatant included positive expression and secretion of the T.gondii protein (SAG1) into the media, indicated by acid phosphatase assay. The characteristics of the SAG1 protein were established using immunofluorescence, immunoblot, ELISA, and mass spectrometry. Conclusion: This study confirms that L. tarentolae effectively can produces and secretes recombinant T. gondii proteins (SAG1 and TgCyc18), offering a potential platform for vaccine development against toxoplasmosis.