Abstract:
Hepatocellular carcinoma (HCC) is a prominent contributor to mortality associated with liver
diseases on a global scale. Hepatitis, liver cirrhosis, hepatocellular carcinoma, and the necessity
for liver transplantation are all significantly influenced by HCV globally. The Flaviviridae family
resulted in around 71 million individuals worldwide being afflicted with chronic HCV infections
in 2020. Direct-Acting Antivirals (DAAs) are one type of treatment for the Hepatitis C Virus
(HCV), But are costly, yet there is a possibility of recurring infection. HCV is characterized by the
presence of 7 distinct genotypes. This study focuses on seven genotypes for the construction of
vaccines and suggested a multi-epitope vaccination. The vaccine utilizes mRNA technology to
replicate the genetic makeup of the HCV virus, imitating its natural genome. The vaccine was
constructed of epitopesthat are connected by linkers and signaling peptides, which are surrounded
by 5' and 3' UTRs. The 6xHis-tag will be affixed to the 5' end for immunocytochemistry. Vaccine
was developed, and its characteristics were analyzed using an in-silico technique. The vaccine was
stabilized using a 5' cap and a 3' poly A tail. The IVT mRNA vaccine was prepared using the
mMACHINE T7 Transcription Kit. The HEK293 cell line was transfected and subsequently
utilized to produce the desired protein. Immunocytochemistry (ICC) results confirmed the
presence of the protein antigen in the HEK293 cells. Our vaccine candidate also produces the
antigen to stimulate a cellular immune response in peripheral blood mononuclear cells (PBMCs).
The in vitro transcribed mRNA demonstrated an elevation in the mRNA levels of both T helper 1
(TH1) response inducer, interleukin-12, and TH1 cytokine interferon gamma, thereby indicating
the production of memory T cells. This study has demonstrated that it holds great potential as a
vaccine for further evaluation against HCV.
