Design and construction of DNA vaccine expressing lectin-like oxidize-LDL receptor-1 (LOX-1) as atherosclerosis vaccine candidate

Atherosclerosis, a chronic inflammatory disease, is counted as one of the deadliest cardiovascular diseases. Lectin-like oxidized LDL receptor-1 (LOX-1) is the major receptor of the oxidized LDL and the inhibition of LOX-1 can prevent atherosclerosis. The study aimed to predict the epitope of the human LOX-1 protein, to construct the DNA LOX-1 vaccine, and to analyze its expression in the mammalian cell line (HeLa). LOX-1 epitopes were predicted by NetMHC 4.0 Server, NetCTLPan 1.1 Server, PickPocket 1.0 Server, and NetMHC Stab 1.0 Server tools. Plasmids contain LOX-1 epitope were transformed into Escherichia coli DH5α. The correct clone carrying LOX-1 epitope was verified by sequencing analysis. Subsequently, plasmids were transfected into HeLa cells using lipotransfection method. The mRNA expression was analyzed using qRT-PCR and the protein expression was analyzed using Western blot method. From epitopes prediction tools, human LOX-1 contains two epitopes, FLYSPWWCLAAATLGV and WLWEDGSPLMPHLFRV. Both genes and protein level expression analyses result showed that the LOX-1 mRNA expression in the transfected HeLa cells were higher compared to that in the non-transfected HeLa cells. Our results suggest that pcDNA3.1-LOX-1 is an atherosclerosis DNA vaccine candidate which is recognized in the mammalian cell expression system.