title: VLP production in Leishmania tarentolae: A novel expression system for purification and assembly of HPV16 L1.
abstract: Viral like particles (VLPs) have been used as immunogen for improvement of preventive vaccines against several viral infections in preclinical and clinical trials. These constructs can stimulate both cellular and humoral immunity. Two prophylactic HPV L1 VLP vaccines known as Gardasil and Cervarix were commercialized worldwide. However, there are main problems for expression and purification of VLPs in eukaryotic expression systems such as baculovirus and yeast leading to high cost of these vaccines. A novel Leishmania protozoan system has been applied to produce different recombinant proteins due to unique properties including generation of similar proteins with mammalian, easy handling, and large-scale culture. In the current study, we developed a novel strategy to produce HPV L1 VLP using stably transfected Leishmania cells. The positive transfectants were analyzed by SDS-PAGE and Western blot analysis. The assembly of purified L1 protein was detected by TEM microscopy. Finally, C57BL/6 mice were immunized by crude VLPs and antibody responses were assessed. The results of electronic microscopy revealed average 55-60 nm for L1 VLP. Furthermore, high IgG1 and IgG2a antibody responses were generated by L1 VLPs in mice similar to L1 VLPs produced in baculovirus-infected insect cells. Regarding the results, the amount of recombinant protein generated by Leishmania was 2-3mg/500 ml media, suggesting further optimization of this system for using in large animals and human.
authors: Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran. Electronic address: azam.bolhassani@yahoo.com.
authors: Azam
authors: A
authors: Bolhassani
authors: Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
authors: Zeinab
authors: Z
authors: Shirbaghaee
authors: Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran.
authors: Elnaz
authors: E
authors: Agi
authors: Biotechnology Research Center, Department of Medical Biotechnology, Pasteur Institute of Iran, Tehran, Iran.
authors: Noushin
authors: N
authors: Davoudi
mesh_terms: D000818:Animals; D000917:Antibody Formation; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology; D005260:Female; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D052162:Human papillomavirus 16 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007114:Immunization; D007891:Leishmania / Q000235:genetics*; D008810:Mice, Inbred C57BL; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000276:immunology; D030361:Papillomavirus Infections / Q000276:immunology / Q000517:prevention & control; D053918:Papillomavirus Vaccines / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D058425:Vaccines, Virus-Like Particle / Q000235:genetics* / Q000276:immunology
chemical_list: D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D053918:Papillomavirus Vaccines; D011994:Recombinant Proteins; D058425:Vaccines, Virus-Like Particle; C063317:L1 protein, Human papillomavirus type 16
keywords: Human papillomavirus; L1; Leishmania expression system; Viral like particle