Clinical and Experimental Vaccine Research

The Korean Vaccine Society (대한백신학회)
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Individual expression and processing of hepatitis C virus E1/E2 epitopes-based DNA vaccine candidate in healthy humans' peripheral blood mononuclear cells

  • Rola Nadeem (Department of Therapeutic Chemistry, Pharmaceutical and Drug Industries Research Institute, National Research Center) ;
  • Amany Sayed Maghraby (Department of Therapeutic Chemistry, Pharmaceutical and Drug Industries Research Institute, National Research Center) ;
  • Dina Nadeem Abd-Elshafy (Immune- and Bio-markers for Infection Research Group, Center of Excellence for Advanced Sciences,National Research Center) ;
  • Ahmed Barakat Barakat (Microbiology department, Faculty of Science, Ain Shams University) ;
  • Mahmoud Mohamed Bahgat (Department of Therapeutic Chemistry, Pharmaceutical and Drug Industries Research Institute, National Research Center)
  • Received : 2022.03.25
  • Accepted : 2022.12.23
  • Published : 2023.01.31
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Abstract

Purpose: The development and study of hepatitis C virus (HCV) vaccine candidates' individualized responses are of great importance. Here we report on an HCV DNA vaccine candidate based on selected envelope (E1/E2) epitopes. Besides, we assessed its expression and processing in human peripheral blood mononuclear cells (PBMCs) and in vivo cellular response in mice. Materials and Methods: HCV E1/E2 DNA construct (EC) was designed. The antigen expression of EC was assayed in PBMCs of five HCV-uninfected donors via a real-time quantitative polymerase chain reaction. Serum samples from 20 HCV antibody-positive patients were used to detect each individual PBMCs expressed antigens via enzyme-linked immunosorbent assay. Two groups, five Swiss albino mice each, were immunized with the EC or a control construct. The absolute count of lymph nodes' CD4+ and CD8+ T-lymphocytes was assessed. Results: Donors' PBMCs showed different levels of EC expression, ranging between 0.83-2.61-fold in four donors, while donor-3 showed 34.53-fold expression. The antigens expressed in PBMCs were significantly reactive to the 20 HCV antibody repertoire (all p=0.0001). All showed comparable reactivity except for donor-3 showing the lowest reactivity level. The absolute count % of the CD4+ T-cell significantly increased in four of the five EC-immunized mice compared to the control group (p=0.03). No significant difference in CD8+ T-cells % was observed (p=0.89). Conclusion: The inter-individual variation in antigen expression and processing dominance was evident, showing independence in individuals' antigen expression and reactivity levels to antibodies. The described vaccine candidate might result in a promising natural immune response with a possibility of CD4+ T-cell early priming.

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References

  1. Chan PP, Levy MT, Shackel N, Davison SA, Prakoso E. Hepatocellular carcinoma incidence post direct-acting antivirals in hepatitis C-related advanced fibrosis/cirrhosis patients in Australia. Hepatobiliary Pancreat Dis Int 2020;19:541-6. 
  2. Itakura J, Kurosaki M, Kakizaki S, et al. Features of resistance-associated substitutions after failure of multiple direct-acting antiviral regimens for hepatitis C. JHEP Rep 2020;2:100138. 
  3. Yun H, Zhao G, Sun X, Shi L. Cost-utility of sofosbuvir/velpatasvir versus other direct-acting antivirals for chronic hepatitis C genotype 1b infection in China. BMJ Open 2020;10:e035224. 
  4. Tarr AW, Khera T, Hueging K, et al. Genetic diversity underlying the envelope glycoproteins of hepatitis C virus: structural and functional consequences and the implications for vaccine design. Viruses 2015;7:3995-4046. 
  5. Grebely J, Prins M, Hellard M, et al. Hepatitis C virus clearance, reinfection, and persistence, with insights from studies of injecting drug users: towards a vaccine. Lancet Infect Dis 2012;12:408-14. 
  6. Schulze Zur Wiesch J, Ciuffreda D, Lewis-Ximenez L, et al. Broadly directed virus-specific CD4+ T cell responses are primed during acute hepatitis C infection, but rapidly disappear from human blood with viral persistence. J Exp Med 2012;209:61-75. 
  7. Filskov J, Andersen P, Agger EM, Bukh J. HCV p7 as a novel vaccine-target inducing multifunctional CD4+ and CD8+ T-cells targeting liver cells expressing the viral antigen. Sci Rep 2019;9:14085. 
  8. Bailey JR, Dowd KA, Snider AE, et al. CD4+ T-cell-dependent reduction in hepatitis C virus-specific neutralizing antibody responses after coinfection with human immunodeficiency virus. J Infect Dis 2015;212:914-23. 
  9. Ahrends T, Busselaar J, Severson TM, et al. CD4+ T cell help creates memory CD8+ T cells with innate and help-independent recall capacities. Nat Commun 2019;10:5531. 
  10. Hobernik D, Bros M. DNA vaccines: how far from clinical use? Int J Mol Sci 2018;19:3605. 
  11. Braathen R, Spang HC, Lindeberg MM, et al. The magnitude and IgG subclass of antibodies elicited by targeted DNA vaccines are influenced by specificity for APC surface molecules. Immunohorizons 2018;2:38-53. 
  12. Villadangos JA, Young L. Antigen-presentation properties of plasmacytoid dendritic cells. Immunity 2008;29:352-61. 
  13. Keck Z, Wang W, Wang Y, et al. Cooperativity in virus neutralization by human monoclonal antibodies to two adjacent regions located at the amino terminus of hepatitis C virus E2 glycoprotein. J Virol 2013;87:37-51. 
  14. Ball JK, Tarr AW, McKeating JA. The past, present and future of neutralizing antibodies for hepatitis C virus. Antiviral Res 2014;105:100-11. 
  15. Bolcic F, Sede M, Moretti F, et al. Analysis of the PKR-eI-F2alpha phosphorylation homology domain (PePHD) of hepatitis C virus genotype 1 in HIV-coinfected patients by ultra-deep pyrosequencing and its relationship to responses to pegylated interferon-ribavirin treatment. Arch Virol 2012;157:703-11. 
  16. Forns X, Payette PJ, Ma X, et al. Vaccination of chimpanzees with plasmid DNA encoding the hepatitis C virus (HCV) envelope E2 protein modified the infection after challenge with homologous monoclonal HCV. Hepatology 2000;32:618-25. 
  17. Alvarez-Lajonchere L, Shoukry NH, Gra B, et al. Immunogenicity of CIGB-230, a therapeutic DNA vaccine preparation, in HCV-chronically infected individuals in a Phase I clinical trial. J Viral Hepat 2009;16:156-67. 
  18. Masavuli MG, Wijesundara DK, Underwood A, et al. A hepatitis C virus DNA vaccine encoding a secreted, oligomerized form of envelope proteins is highly immunogenic and elicits neutralizing antibodies in vaccinated mice. Front Immunol 2019;10:1145. 
  19. Alemu EY, Carl JW Jr, Corrada Bravo H, Hannenhalli S. Determinants of expression variability. Nucleic Acids Res 2014;42:3503-14. 
  20. Kennedy RB, Ovsyannikova IG, Palese P, Poland GA. Current challenges in vaccinology. Front Immunol 2020;11:1181. 
  21. Wasenius VM, Hemmer S, Kettunen E, Knuutila S, Franssila K, Joensuu H. Hepatocyte growth factor receptor, matrix metalloproteinase-11, tissue inhibitor of metalloproteinase-1, and fibronectin are up-regulated in papillary thyroid carcinoma: a cDNA and tissue microarray study. Clin Cancer Res 2003;9:68-75. 
  22. De Winter JC. Using the Student's t-test with extremely small sample sizes. Pract Assess Res Eval 2013;18:10. 
  23. Meunier JC, Russell RS, Goossens V, et al. Isolation and characterization of broadly neutralizing human monoclonal antibodies to the e1 glycoprotein of hepatitis C virus. J Virol 2008;82:966-73. 
  24. Morin TJ, Broering TJ, Leav BA, et al. Human monoclonal antibody HCV1 effectively prevents and treats HCV infection in chimpanzees. PLoS Pathog 2012;8:e1002895. 
  25. Keck ZY, Angus AG, Wang W, et al. Non-random escape pathways from a broadly neutralizing human monoclonal antibody map to a highly conserved region on the hepatitis C virus E2 glycoprotein encompassing amino acids 412-423. PLoS Pathog 2014;10:e1004297. 
  26. Keck ZY, Saha A, Xia J, et al. Mapping a region of hepatitis C virus E2 that is responsible for escape from neutralizing antibodies and a core CD81-binding region that does not tolerate neutralization escape mutations. J Virol 2011;85:10451-63. 
  27. Owsianka AM, Timms JM, Tarr AW, et al. Identification of conserved residues in the E2 envelope glycoprotein of the hepatitis C virus that are critical for CD81 binding. J Virol 2006;80:8695-704. 
  28. Buccitelli C, Selbach M. mRNAs, proteins and the emerging principles of gene expression control. Nat Rev Genet 2020;21:630-44. 
  29. Franco LH, Wowk PF, Silva CL, et al. A DNA vaccine against tuberculosis based on the 65 kDa heat-shock protein differentially activates human macrophages and dendritic cells. Genet Vaccines Ther 2008;6:3. 
  30. Cresswell P. Assembly, transport, and function of MHC class II molecules. Annu Rev Immunol 1994;12:259-93. 
  31. Sadegh-Nasseri S, Kim A. Selection of immunodominant epitopes during antigen processing is hierarchical. Mol Immunol 2019;113:115-9. 
  32. Ascough S, Ingram RJ, Chu KK, et al. CD4+ T cells targeting dominant and cryptic epitopes from Bacillus anthracis lethal factor. Front Microbiol 2016;6:1506. 
  33. Dai G, Steede NK, Landry SJ. Allocation of helper T-cell epitope immunodominance according to three-dimensional structure in the human immunodeficiency virus type I envelope glycoprotein gp120. J Biol Chem 2001;276:41913-20. 
  34. Sandberg JK, Glas R. Antigen processing. Hoboken (NJ): Encyclopedia of Life Science/John Wiley & Sons; 2001. 
  35. Chang S, Kodys K, Szabo G. Impaired expression and function of toll-like receptor 7 in hepatitis C virus infection in human hepatoma cells. Hepatology 2010;51:35-42. 
  36. Wang J, Symul L, Yeung J, et al. Circadian clock-dependent and -independent posttranscriptional regulation underlies temporal mRNA accumulation in mouse liver. Proc Natl Acad Sci U S A 2018;115:E1916-25. 
  37. Garcia-Arriaza J, Esteban M. Enhancing poxvirus vectors vaccine immunogenicity. Hum Vaccin Immunother 2014;10:2235-44. 
  38. Frey SE, Houghton M, Coates S, et al. Safety and immunogenicity of HCV E1E2 vaccine adjuvanted with MF59 administered to healthy adults. Vaccine 2010;28:6367-73.