Expression of Hepatitis B Surface Antigen in Coleus forskohlii

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Research Paper 01/07/2016
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Expression of Hepatitis B Surface Antigen in Coleus forskohlii

Neha Guleria, Paramanahally Hanumanthe Gowda Ramanjini Gowda, Satish Kumar Kariyaiah, Renu Pasricha
Int. J. Biomol. & Biomed.5( 1), 29-39, July 2016.
Certificate: IJBB 2016 [Generate Certificate]

Abstract

Hepatitis B, a common viral infection, affecting millions of people every year, leads to disability and death. It has become one of the alarming diseases in the world. Vaccination is the best possible way to prevent this deadly viral infection and its consequences. Unaffordability of the mass vaccination program due to low health budgets especially in developing countries have demanded the economical, effective and easily available vaccine production against hepatitis B virus. The expression of subunit vaccines and recombinant proteins in plants is a convenient, safe and potentially economical platform technology. Hence, development of a plant-based vaccine could be promising. Therefore, the present investigation focused on expression and large- scale production of hepatitis B surface antigen (HBsAg) in Coleus forskohlii for the development of vaccine. Eight transformed C. forskohlii plants were generated via Agrobacterium -mediated transformation method. The integration of 681bp of HBsAg gene into the plant genome was confirmed using PCR. SDS-PAGE showed the presence of ~48kDa dimer and ~24kDa monomer form of HBsAg protein and the expression of recombinant protein was further confirmed by western blot. C. forskohlii expressed HBs Ag was recorded 260µg/g leaf fresh weight as measured by ELISA. Transformed plants of HBsAg showed the accumulation of Virus Like Particles of 22 nm size using transmission electron microscopy. This study offers a great potential for the large -scale production of hepatitis B vaccine in C. forskohlii which provides a strategy for contributing a means to achieve global immunization for the hepatitis B prevention and eradication.

VIEWS 24

Ahmad K. 2014. Molecular farming: strategies, expression systems and bio-safety considerations. Czech Journal of Genetics and Plant Breeding 50,1-10.

Balen B, Rasol MK. 2007. N-Glycosylation of Recombinant Therapeutic Glycoproteins in Plant Systems. Food Technology and Biotechnology 45,1–10.

Boby BU, Bagyaraj DJ. 2003. Biological control of root- rot of Coleus forskohlii Briq. using microbial inoculants. World Journal of Microbiology and Biotechnology19,175-180.

CzyzM, Dembczynski R, Marecik R, Wojas-Turek J, Milczarek M, Pajtasz-Piasecka E, Wietrzyk J, Pniewski T. 2014. Freeze-Drying of Plant Tissue Containing HBV Surface Antigen for the Oral Vaccine against Hepatitis B. [published online ahead of print October 12, 2014]. BioMed Research International doi:10.1155/2014/485689.

De la Riva GA, Cabrera GJ, Padron RV, Pardo CA.1998. Agrobacterium tumefaciens: a natural tool for plant transformation. Electronic Journal of Biotechnology doi: 10.2225/Vol.1-issue 3.

Doyle JJ, Doyle JL.1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin 19,11-15.

Dwarakaprasad AA, Rao S. 2013. Virus like particles as vaccines. International Journal for Pharmaceutical Research Scholars 2, 512-544.

Ehsani P, Khabiri A, Domansky NN.1997. Polypeptides of hepatitis B surface antigen produced in transgenic potato. Gene 190, 107–111.

ElkholySF, Roba MI, Ahmed B, Atef SS, Magdy AM. 2009. Expression of Hepatitis B surface antigen (HBsAg) gene in transgenic banana (Musa sp). Arabian Journal of Biotechnology 12, 291-302.

Gao Y, Ma Y, Li M, Cheng T, Li SW, Zhang J, Xia NS. 2003. Oral immunization of animals with transgenic cherrytomatillo expressing HBsAg. World Journal of Gastroenterology 9, 996–1002.

Goldstein ST, Zhou FHadler SCBell BPMast EE, Margolis HS. 2005. A mathematical model to estimate global hepatitis B disease burden and vacci-nation impact. International Journal of Epidemi-ology 34, 1392-1339.

Greiner VJ, Ronzon F, Larquet E, Desbat B, Esteves C, Bonvin J, Greco F, Manin C, Klymchenko AS, Mely Y. 2012. The structure of HBsAg particles is not modified upon their adsorption on alu­minium hydroxide gel. Vaccine 30, 5240–5245.

Guleria N, Gowda Ramanjini PH. 2015. An efficient regeneration and genetic transformation protocol of Coleus forskohlii using Biolistic gun. International Journal of Agriculture, Environment and Biotechnology 8, 227-235.

Hu I, Metz S, Chay C, Zhou HP, Biest N, Chen G, Cheng M, Feng X, Radionenko M, Lu F, Fry J. 2003. Agrobacterium– mediated large scale transformation of wheat using glyphosate selection. Plant Cell Reports 21,1010-1019.

Hyun SK, Jae HJ, Kyung JL, Kisung K. 2014. N-Glycosylation Modification of Plant-Derived Virus-Like Particles: An Application in Vaccines. [published online ahead of print May 25, 2014]. BioMed Research International doi.org/10.1155/2014/249519es.

Jadwiga C, Inga S, Ewa S. 2014. Virus-like particles as vaccine .Acta Biochimica Polonica 61,531–539.

Kapusta J, Modelska A, Figlerowicz M, Pniewski T, Letellier M, Lisowa O, Yusibov V, Koprowski H, Plucienniczak A Legocki AB. 1999. A plant-derived edible vaccine against hepatitis B virus. Federation of American Societies for Experi-mental Biology 13,1796-1999.

Khan MY, Saleh A, Vimal K, Rajkumar S. 2009. Recent advances in medicinal plant biotechnology. Indian Journal of Biotechnology 8, 9-22.

Khin ML, Gowda Ramanjini PH, Chandrashekar S, Gowda TKS, Vasundhara M, Nagesha N, Virupaksha GUP, Sreenevasa S, Manjunatha T. 2006. In vitro regeneration and transformation of Coleus forskohlii with glucanase-chitinase gene. Journal of Non-Timber Forest Products 13,193-197.

Kong Q, Richter L, Yang YF, Arntzen CJ, Mason HS, Thanavala Y. 2001. Oral immunization with hepatitis B surface antigen expressed in transgenic plants. Proceedings of National Academy of Sciences USA 98, 11539–11544.

Li T, Jing Kuan S, Zhao Hua L, Qing L. 2011. Transformation of HBsAg (Hepatitis B Surface Antigen) Gene into Tomato Mediated by Agrobacterium tumefa-ciens. Czech Journal of Genetics and Plant Breeding 47, 69–77.

Lou XM, Yao QH,  Zhang Z, Peng R H, Xiong SA, Wang HK. 2007. Expression of the human hepatitis B virus large surface antigen gene in transgenic tomato plants. Clinical Vaccine Immunology 4, 464–469.

Ma JKC, Drake P, Christou P. 2003. The production of recombinant pharmaceutical proteins in plants. Nature Reviews Genetics 4, 794-805.

Mason HS, Arntzen CJ. 1995. Transgenic plants as vaccine production systems. Trends in Biotechnology 13, 388–392.

Mason HS, Lam DM, Arntzen CJ. 1992. Expression of hepatitis B surface antigen in transgenic plants. Proceedings of National Academy of Sciences USA 89, 11745–11749.

Merle C, Perret S, Lacour T, Jonval V, Hudaverdian S, Garrone R, Ruggiero F, Theisen M. 2002. Hydroxylated human homotrimeric collagen I in Agrobacterium tumefaciens-mediated transient exp-ression and in transgenic tobacco plant. Federation of European Biochemical Societies Letters 515, 114–118.

Mishiro S, Imai M, Takahashi K, Machida A, Gotanda T, Miyakawa Y, Mayumi M. 1980. A 49000-dalton polypeptide bearing all antigenic determinants and full immunogenicity of 22-nm hepatitis B surface antigen particles. Journal of Immunology 124, 1589–1593.

Murashige J, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco cultures. Plant Physiology 15, 473-497.

Natasha K, Stephen J. Streatfield, VY. 2012. Virus-like particles as a highly efficient vaccine platform: Diversity of targets and production systems and advances in clinical development. Vaccine 31, 58-83.

Peeters K, De Wilde V, De Jaeger C, Angenon GG, Depicker A.2001. Production of antibodies and antibody fragments in plants. Vaccine 19, 2756–2761.

Pniewski T. The twenty year story of a plant-based vaccine against hepatitis B: Stagnation or Promising Prospects? International Journal of Molecular Sciences 14, 1978-1998.

Richter LJ, Thanavala Y, Arntzen CJ, Mason HS. 2000. Production of hepatitis B surface antigen in transgenic plants for oral immunization. Nature Biotechnology 18, 1167–1171.

Roldao A, Mellado MCM, Castilho LR, Carrondo MJT, Alves PM. 2010. Virus like particles in vaccine development. Expert Review of Vaccines 9, 1149-1176.

Rybicki EP. 2014. Plant based vaccines against viruses. Virology Journal 11, 205.

RukavtsovaElena B, Natalya V Rudenko, Elena N Puchko, Natalya SZakharchenko, YaroslavI Buryanov. 2015. Study of the immunogenicity of hepatitis B surface antigen synthesized in transgenic potato plants with increased biosafety. Journal of Biotechnology 203, 84–88.

Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning: A Laboratory Manual (2nd edition). Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

Santi L, Zhong H, Mason H. 2006. Virus like particles production in green plants. Methods 40, 66-76.

Smith ML, Mason HS, Shuler ML. 2002b. Hepatitis B surface antigen (HBsAg) expression in plant cell culture: kinetics of antigen accumulation in batch culture and its intracellular form. Biotechnology Bioengineering 80, 812–822.

Smith ML, Richter L, Arntzen CJ, Shuler ML, Mason HS. 2003. Structural characterization of plant-derived hepatitis B surface antigen employed in oral immunization studies. Vaccine 21, 4011–4021.

Sojikul P, Buehner N, Mason HS. 2003. A plant signal peptide–hepatitis B surface antigen fusion protein with enhanced stability and immunogenicity expressed in plant cells. Proceedings of National Academy of Sciences USA 100, 2209–2214.

Thomas DR, Claire AP, Amrita M, Amanda MW. 2011. Evolution of plant –made pharmaceuticals. International Journal of Molecular Sciences 12, 3220-3236.

Wampler DE, Lehman ED, Boger J, McAleer WJ, Scolnick EM. 1985. Multiple chemical forms of hepatitis B surface antigen produced in yeast. Proceedings of National Academy of Sciences USA 82, 6830–6834.

WHO. 2015. Hepatitis B. Available at http://www. who.int/mediacentre/factsheets/. March, 2015. Acce-ssed on April 2, 2015.

Xu J, Dolan MC, Medrano G, Cramer CL, Weathers PJ. 2012. Green factory: plants as bioproduction platforms for recombinant proteins. Biotechnology Advances 30, 1171-1184.

Yogendra KN, Ramanjini Gowda PH, Sandesh HS, Raghavendra G, Asha VN, Ningaraju TM, Deepak N. 2009. Production and Characterization of Hepatitis B Recombinant Vaccine in Tobacco (Nicotiana tobaccum cv. ‘Kanchun’).Transgenic Plant Journal 3, 97-101.

YogendraKN, Raghavendra G, Ramanjini Gowda PH. 2013. Stability and inheritance analysis of transgenic tobacco expressing Hepatitis B surface antigen. Australian Journal of Crop Sciences 7, 1010-1015.

Zhong H, Galina E, Bryan J, Maloneyc NB, Charles JA, Yasmin T, Mason HS. 2005. Virus-like particle expression and assembly in plants: hepatitis B and Norwalk viruses. Vaccine 23, 1851–1858.

Zhong H, Kate Le P, Galina E, Yasmin T, Mason HS. 2008. High-yield rapid production of hepatitis B surface antigen in plant leaf by a viral expression system. Plant Biotechnology Journal 6, 202–209.

Zhong H, Mason HS. 2004. Conformational analysis of hepatitis B surface antigen fusions in an Agrobacterium-mediated transient expression system. Plant Biotechnology Journal 2, 241–249.