The role of type I interferon in the immunobiology of chikungunya virus

Wilson, Jane Amelia Clare (2015). The role of type I interferon in the immunobiology of chikungunya virus PhD Thesis, School of Medicine, The University of Queensland. doi:10.14264/uql.2016.11

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Author Wilson, Jane Amelia Clare
Thesis Title The role of type I interferon in the immunobiology of chikungunya virus
School, Centre or Institute School of Medicine
Institution The University of Queensland
DOI 10.14264/uql.2016.11
Publication date 2015-12-18
Thesis type PhD Thesis
Supervisor Andreas Suhrbier
Wayne A Schroder
Alex Khromykh
Total pages 255
Language eng
Subjects 110707 Innate Immunity
110804 Medical Virology
Formatted abstract
Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that can cause explosive outbreaks of a febrile, arthritic/arthralgic disease usually lasting weeks to months, and in rare cases, more than a year. In 2004, the largest ever CHIKV outbreak began in Kenya, spreading to islands of the Indian Ocean, India, South East Asia and major outbreaks have recently occurred in the South Pacific Islands and the Caribbean. The host type I interferon (IFN) response is crucial for effective control of CHIKV infection. Herein, the dynamics, source and responses generated by the type I IFNs following CHIKV infection were investigated.

Interferon regulatory factors 3 (IRF3) and IRF7 are key transcription factors for the type I IFN response. While CHIKV infection of wild-type mice is non-lethal, infection of mice deficient in both IRF3 and IRF7 (IRF3/7-/-) resulted in mortality, illustrating that these factors are essential for protection. Using knockout mice for the adaptor molecules upstream of IRF3 and 7, IPS1 was found to be the most important for type I IFN production, with TRIF and MyD88 also contributing to the response. Mortality in IRF3/7-/- mice was also associated with type I IFN suppression of pathological levels of IFNγ and haemorrhagic shock.

Heterozygous reporter mice, in which eGFP was expressed under the control of either the IFNβ or the IFNα6 promoter on one chromosome, were employed to try and identify the cellular source of type I IFN production following CHIVK infection. However, eGFP production was found to be insufficient for detection, limiting the utility of this approach. Investigations of type I IFN production in tissues using RT-PCR revealed a strong correlation between type I IFN induction and viral tissue titres, with feet and lymph nodes producing the most robust type I IFN responses.

RNASeq was undertaken to examine, in detail, the nature of the type I IFN responses generated in the feet and inguinal lymph nodes of mice following CHIKV infection at days 2 and 7 post-infection. High quality sequencing data was generated from III host and viral poly-adenylated RNA, permitting investigation of low level gene transcription from both sources, and the identification of novel genes/transcripts, in addition to analysis of differential gene expression.

Investigation of host transcriptional activity revealed a type I IFN dominated immune response, in spite of a low induction of type I IFN mRNA transcripts at day 2 and an absence of type I IFN mRNA induction on day 7. Many type I IFN-regulated genes, including IRF7, remained up-regulated in the feet at day 7, suggesting type I IFN-independent mechanisms for maintenance of type I IFN-regulated gene expression. Six novel IRF3 isoforms and a potentially novel protein coding gene were also identified.

RNA-Seq analysis of CHIKV transcriptional activity reflected the known CHIKV replication kinetics, in which virus replicates in the joints, and spreads to the draining lymph nodes, with viral titres peaking at day 2 and largely resolving at day 7. Assessment of mutations within the CHIKV genome showed an accumulation with time, with error accumulation higher in the lymph nodes than in the feet. These studies represent the first deep sequencing analysis of CHIKV genomes in tissues.

The findings in this thesis substantially add to our knowledge of the role and function of the immune response after CHIKV, illustrating for the first time that type I IFNs protect against haemorrhagic shock and that expression of type I IFN inducible genes is maintained well after type I IFN induction has waned.
Keyword Chikungunya virus
Type I interferon
Innate Immunity
Infectious disease

Document type: Thesis
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Created: Fri, 11 Dec 2015, 11:17:20 EST by Jane Wilson on behalf of University of Queensland Graduate School