Codon optimization and subcellular targeting were studied with the aim to increase the expression levels of the SAG178-322 antigen of Toxoplasma gondii in tobacco leaves. The expression of the tobacco-optimized and native versions of the SAG1 gene was explored by transient expression from the Agrobacterium tumefaciens binary expression vector, which allows targeting the recombinant protein to the endoplasmic reticulum (ER) and the apoplast.
Finally, mice were subcutaneously and orally immunized with leaf extracts-SAG1 and the strategy of prime boost with rSAG1 expressed in Escherichia coli was used to optimize the oral immunization with leaf extracts-SAG1.
Results: Leaves agroinfiltrated with an unmodified SAG1 gene accumulated 5- to 10-fold more than leaves agroinfiltrated with a codon-optimized SAG1 gene. ER localization allowed the accumulation of higher levels of native SAG1.
However, no significant differences were observed between the mRNA accumulations of the different versions of SAG1. Subcutaneous immunization with leaf extracts-SAG1 (SAG1) protected mice against an oral challenge with a non-lethal cyst dose, and this effect could be associated with the secretion of significant levels of IFN-.
The protection was increased when mice were ID boosted with rSAG1 (SAG1+boost). This group elicited a significant Th1 humoral and cellular immune response characterized by high levels of IFN-.
In an oral immunization assay, the SAG1+boost group showed a significantly lower brain cyst burden compared to the rest of the groups.
Conclusion: Transient agroinfiltration was useful for the expression of all of the recombinant proteins tested. Our results support the usefulness of endoplasmic reticulum signal peptides in enhancing the production of recombinant proteins meant for use as vaccines.
The results showed that this plant-produced protein has potential for use as vaccine and provides a potential means for protecting humans and animals against toxoplasmosis.
Author: Melina Laguia-BecherValentina MartinMauricio KraemerMariana CoriglianoMaria YaconoAlejandra GoldmanMarina Clemente
Credits/Source: BMC Biotechnology 2010, 10:52
A vaccine for T. Gondii could be a valuable tool in addressing some of the negative impacts that Toxo can have on humans. Tobacco leaves as a source for this immunization strategy is highly scalable and is something that we could produce domestically for export. The reduction of cysts in the brains of the mice directly affects the efficacy of Toxo to modify the behavior of the host animals.
Note: I would greatly appreciate anyone with sufficient scientific knowledge to parse the article into more clear laymans terms. I'm sure there are fascinating tidbits in this article that I'm completely missing.
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