Production of a plant-derived immunogenic protein targeting ApoB100 and CETP: toward a plant-based atherosclerosis vaccine.

In an effort to initiate the development of a plant-based vaccination model against atherosclerosis, a cholera toxin B subunit (CTB)-based chimeric protein was designed to target both ApoB100 and CETP epitopes associated with immunotherapeutic effects in atherosclerosis. Epitopes were fused at the C-terminus of CTB to yield a protein called CTB:p210:CETPe. A synthetic gene coding for CTB:p210:CETPe was successfully transferred to tobacco plants with no phenotypic alterations. Plant-derived CTB:p210:CETPe was expressed and assembled in the pentameric form. This protein retained the target antigenic determinants, as revealed by GM1-ELISA and Western blot analyses. Higher expresser lines reached recombinant protein accumulation levels up to 10 µg/g fresh weight in leaf tissues and these lines carry a single insertion of the transgene as determined by qPCR. Moreover, when subcutaneously administered, the biomass from these CTB:p210:CETPe-producing plants was able to elicit humoral responses in mice against both ApoB100 and CETP epitopes and human serum proteins. These findings evidenced for the first time that atherosclerosis-related epitopes can be expressed in plants retaining immunogenicity, which opens a new path in the molecular farming field for the development of vaccines against atherosclerosis.

Autoantibodies to oxidized low density lipoprotein in relation with coronary artery disease.

The relationship between autoantibodies (autoAbs) to oxidized LDL (oxLDL) and coronary artery disease (CAD) remains controversial. IgM and IgG autoAbs to oxLDL and 1-palmitoyl-2 (5'-oxo-valeroyl)-sn-glycero-3-phosphorylcholine (POVPC), as well as the levels of non modified or modified ApoB-100 immune complexes (ICs), were measured in twenty patients undergoing clinically indicated coronary angiography, and in ten young healthy volunteer sera. The levels of IgM autoAbs to oxLDL did not differ between no CAD patients and healthy subjects, but the levels of these autoAbs were significantly higher in no CAD patients and healthy subjects in comparison with CAD patients. There was not difference in the levels of IgM anti-ApoB-100 ICs between both groups of patients. In contrast, the levels of ICs formed by IgM autoAbs and oxidative modified ApoB-100 were lower in patients with CAD than in patients without CAD. No differences were observed in the levels of autoAbs to POVPC among the groups. In conclusion, our results showed that the level of circulating oxLDL IgM autoAbs was lower in CAD patients than in no CAD patients, supporting the hypothesis that this kind of autoAbs might be inversely associated with the presence of atherosclerosis.