title: Subunit Protein Vaccine Delivery System for Tuberculosis Based on Hepatitis B Virus Core VLP (HBc-VLP) Particles.
abstract: Despite the development of modern medicine, tuberculosis (TB), caused by the pathogenic bacterium, Mycobacterium tuberculosis (Mtb), remains one of the deadliest diseases. This bacterium can lay dormant in individuals and get activated when immunity goes down and has also shown considerable prowess in mutating into drug resistant forms. The global emergence of such drug resistant Mtb and the lack of efficacy of Bacille Calmette Guérin (BCG), the only vaccine available so far, have resulted in a situation which cries out for a safe and effective tuberculosis vaccine.Number of different strategies has been used for developing new anti-TB vaccines and several protective antigens have been identified so far. One strategy, the use of protein subunits, has the potential to develop into a powerful tuberculosis vaccine, not only because of its efficacy and safety, but also because they are economical. The proper delivery of protein subunit vaccines with adjuvants or novel delivery systems is necessary for inducing protective immune responses. The available adjuvants or delivery systems are inadequate for generating such a response. In the present method, we have constructed a vaccine delivery system for tuberculosis based on Virus-Like Particles (VLPs). Hepatitis B Virus core antigen gene was recombinantly modified using Overlap Extension PCR (OEPCR). The final construct was designed to express HBc-VLP carrying external antigen (fusion VLP). Mycobacterium tuberculosis antigen CFP-10 was used for the construction of fusion VLP. The recombinant gene for the construct was cloned into a pET expression system and transformed into E. coli BL21(DE3) and induced with IPTG to express the protein. The fusion protein was purified using the Histidine tag and allowed to form VLPs. The preformed VLPs were purified by sucrose density gradient centrifugation. The VLPs were characterized using Transmission Electron Microscopy (TEM).
authors.affiliation: Mycobacteria Research, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, 695 014, Kerala, India.
authors.forename: Dhananjayan
authors.initials: D
authors.lastname: Dhanasooraj
authors.affiliation: Mycobacteria Research, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, 695 014, Kerala, India.
authors.forename: R Ajay
authors.initials: RA
authors.lastname: Kumar
authors.affiliation: Mycobacteria Research, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, 695 014, Kerala, India. smundayoor@rgcb.res.in.
authors.forename: Sathish
authors.initials: S
authors.lastname: Mundayoor
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D006515:Hepatitis B virus / Q000737:chemistry*; D009169:Mycobacterium tuberculosis / Q000276:immunology; D021141:Nucleic Acid Amplification Techniques; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic; D022223:Vaccines, Subunit / Q000235:genetics*; D058425:Vaccines, Virus-Like Particle / Q000235:genetics*
chemical_list: D022223:Vaccines, Subunit; D058425:Vaccines, Virus-Like Particle
keywords: Adjuvant; CFP 10; HBc-VLP; Mycobacterium tuberculosis; Subunit vaccine; Virus-like particle