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P10.22 Engineering human rhinovirus serotype-a1 as a vaccine vector
  1. K Tomusange1,
  2. W Yu1,
  3. A Suhrbier2,
  4. D Wijesundara1,
  5. B Grubor1,
  6. E Gowans1
  1. 1Virology Laboratory, Basil Hetzel Institute, Discipline of Surgery, University of Adelaide, Adelaide, South Australia, Australia
  2. 2QIMR Berghofer Medical Research Institute, Brisbane, Queensland, Australia


Introduction Vaccination is the optimal long-term solution to the human immuno-defficiency virus (HIV) pandemic. A majority of new HIV infections result from mucosal transmission, highlighting the need for novel vaccines that primarily generate mucosal immunity. Like HIV, human rhinovirus serotype-A1 (HRV-A1, the common cold virus) is primarily transmitted via mucosal surfaces. This makes HRV-A1 a potential vector for mucosally targeted vaccines to generate robust protection against HIV at the vaginal and other distant mucosal surfaces, and systemically.

Methods Using recombinant technology, we inserted discrete overlapping fragments of HIV gag and full-length tat into the junction between genes that encode HRV-A1 structural and non-structural proteins to generate recombinant HRV-A1s (rHRV-A1s) encoding HIV Gag and Tat proteins.

Results Transfection of H1-HeLa cells with rHRV-A1s transcripts yielded infectious and replication-competent rHRV-A1s with similar growth characteristics as wildtype HRV-A1. We also confirmed that cells infected with rHRV-A1s stably expressed Gag and Tat (beyond 5 passages), of the correct sizes and mainly localised in the cytoplasm as revealed by western blot assay, reverse-transcription polymerase chain reaction and immunofluorescence. These results have been recently accepted for publication in Virus Research Journal (reference number: VIRUS96578, April 2015).

Conclusion To the best of our knowledge, this is the first time replication-competent and stable HRV-A1 vectors have been generated. The individual rHRV-A1gag/tat generated in this study have been mixed into a cocktail vaccine and administered intranasally to female Balb/C mice to evaluate its immunogenicity (animal experiments currently on-going). The protective efficacy of the resultant HIV-Gag-specific cell-mediated and Tat humoral responses will be documented in mice challenged intravaginally with chimeric rodent ecotropic murine leukaemia virus (EcoHIV). EcoHIV was developed by replacing the coding region of glycoprotein-120 (gp-120) of HIV strain NL4–3 with gp80 of EcoHIV to ensure that the chimeric virus infected murine cells only.

Disclosure of interest statement This work was supported by a grant from The Hospital Research Foundation (THRF), Woodville, South Australia and grant number APP1026293 from the National Health and Medical Research Council (NHMRC) of Australia. TK is a post-graduate Fellow supported by THRF while SA and GE are Senior Research Fellows of the NHMRC. The authors declare no financial or commercial conflict of interest.

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