BIO JOHOR 2014 4TH JOHOR BIOTECHNOLOGY CONFERENCE & EXHIBITION 26th -27th August 2014 | The Puteri Pacific, Johor Bahru, Johor, Malaysia EXTENDED ABSTRACT FORMAT 1. Title: Bold, Arial font size 11, capital letters, centered. 2. Author(s): Full name of authors, underline the name of presenter (for oral) and corresponding author (for poster); font size 11. 3. Author affiliation(s): Full addresses of each author in italics, Arial font size 10, provide email address of corresponding author. If more than one institution, indicate the address of each author numerically in superscript. 4. Language: British English 5. Text should be divided into: Abstract, Introduction, Materials and Methods, Results and Discussion, Acknowledgement and References. 6. Margin: 1” left, top, bottom and right borders with single spacing (1.0). 7. Abstract: One paragraph, Arial font size 11, justified, do not include figures/tables/references and not more than 250 words. 8. Extended abstract should be limited to not more than 4 pages in Arial font size 11, including figures and tables. Figures must be of high quality. BIO JOHOR 2014, Johor Biotechnology Conference and Exhibition, August 25-27, 2014, Persada Johor, Malaysia. THE USE OF RHDV VIRUS-LIKE PARTICLES AS AN IMMUNOTHERAPY FOR HUMAN PAPILLOMAVIRUS TYPE 16 TUMOURS Khairunadwa Jemon1, Merilyn Hibma1, Vernon Ward1, Sarah Young2 1 2 Department of Microbiology and Immunology and Department of Pathology, University of Otago, New Zealand. Corresponding author email: [email protected] Abstract – These instructions give you basic guidelines for preparing papers for conference proceedings. Keywords – Sustainability Social; Responsibility; INTRODUCTION Human papillomavirus (HPV) is the primary etiological agent of cervical cancer. The consistent over-expression of the HPV16 E6 and E7 oncogenes is required to induce and maintain the transformed phenotype of cervical cancer cells. Better therapeutic strategies are required for the treatment of pre-existing, cancer-causing, HPV infections. It has been previously shown that virus-like particles (VLP) made by expressing the capsid protein of rabbit hemorrhagic disease virus (RHDV) are capable of delivering a tumour antigen, leading to significant antitumour effect against melanoma1. This suggests that these VLP may also be effective in delivering other antigens to stimulate an immune response against other types of tumour. 2-3 days using digital callipers, up to a maximum of 90 days, or until tumours had reached the maximum allowable size of 150 mm2. RESULTS Fig. 2: Establishing in vivo tumour model in C57BL/6 mouse. Mice were inoculated with (A) 5 × 104 cells/mouse or (B) 1×105 cells/mouse. It was decided to use the low dose, 5×104 cells/mouse for the subsequent experiments. AIM: To ascertain whether RHDV VLP expressing E6 have the ability to delay or prevent the growth of an HPV16 E6 and E7 expressing tumour. METHODS Fig. 1: Schematic diagram for the in vivo tumour treatment experiment. C57BL/6 mice were inoculated with 5 x 104 TC-1 cells expressing HPV16 E6 and E72. Five days after tumour inoculation, mice were administered with 170 μg/mouse of PC61 (anti-CD25 monoclonal antibody) to deplete regulatory T cells. Mice were vaccinated with RHDV VLP16 E6 with PADRE peptide, a universal pan DR epitope3, twice with a one week interval. Control groups were vaccinated either with VLP with PADRE or PBS only. Mice were monitored and tumours were measured every Fig. 3: In vivo tumour treatment experiments. Mice were vaccinated with (A) VLP-16 E6 PADRE or (B) VLP-PADRE; (C) PBS. Line graphs depicting the mean tumour area BIO JOHOR 2014, Johor Biotechnology Conference and Exhibition, August 25-27, 2014, Persada Johor, Malaysia. (mm2) over time (mean ± S.E.M). Tumour sizes in the mice were monitored and measured every 2 days. DISCUSSION Summary of results: 1. Some reduction in tumour size was observed in the tumour positive mice vaccinated with VLP16 E6 PADRE compared to the control groups; VLP PADRE or PBS (Fig. 3). 2. Tumour positive mice treated with PC61 antibody showed some reduction in tumour size, indicating that CD25+ regulatory T cell depletion improved the efficacy of this vaccine (Fig. 3). Although no statistical difference was measured, these results provide some indication that the VLP16 E6 PADRE used in this experiment has an antitumour effect against an HPV16 E6 and E7 expressing tumour. A limitation of this experiment was that only around half of the control unimmunised mice developed tumours. Since this experiment, we have reselected the TC-1 cells with G418 and hygromycin B. There was significant cell death under selection, suggesting that there had been outgrowth of cells that did not express the transgenes. This experiment will be repeated using selected cells to confirm the efficacy of these VLP as a vaccine against tumour expressing HPV16 E6 and E7. ACKNOWLEDGEMENTS We would like to thank Health Research Council of New Zealand for funding. REFERENCES Alexander, J., Sidney, J., Southwood, S., Ruppert, J., Oseroff, C., Maewal, A., Snoke, K., Serra, H.M., Kubo, R.T., Sette, A., and Grey, H.M. (1994). Development of high potency universal DR-Restricted helper epitopes by modification of high affinity DR-blocking peptides. Immunity 1: 751-761. Lin, K.Y., Guarnieri, F.G., StaveleyO'Carroll, K.F., Levitsky, H.I., August, J.T., Pardoll, D.M., Wu, T.C. (1996). Treatment of established tumors with a novel vaccine that enhances major histocompatibility class II presentation of tumor antigen. Cancer Res. 56: 21-26. Peacey, M., Wilson, S., Perret, R., Ronchese, F., Ward, V.K., Young, V., Young, S.L. and Baird, M. (2008). Viruslike particles from rabbit hemorrhagic virus can induce an anti-tumour response. Vaccine 26: 5334-5337.