Defining the intrinsic and innate immune response to UV irradiated HSV-1 infection

Iliev, Victor (2022) Defining the intrinsic and innate immune response to UV irradiated HSV-1 infection. PhD thesis, University of Glasgow.

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Since the late 1960s UV irradiation of Herpesviruses has been used to study various aspects of cell biology, including the cellular immune response to virus infection. Previous studies have demonstrated that UV irradiation of herpesvirus virions strongly stimulates the type I and II IFN response to viral infection. However, the spatial and temporal kinetics of recruitment of immune signalling actuated in response to UV irradiated virus infection have remained poorly understood, due to limitations of viral DNA (vDNA) detection. Utilizing 5-Ethynyl-2’ deoxycytidine (EdC) labelling of HSV-1 DNA in combination with click chemistry, we were able to directly visualize vDNA and characterize the recruitment of host immune factors to infecting wild type (WT) or UV irradiated viral genomes. UV irradiation ofHSV-1 virions induced the premature cytoplasmic release of vDNA in a microtubule-dependent manner. An increase in vDNA thymine dimer formation was observed that restricted genome decompacting once released from the capsid. HSV-1 genomes in the cytoplasm exhibited an enhanced recruitment of cGAS and STING following UV irradiation compared to non-UV irradiated cytoplasmic genomes. Depletion of cGAS led to the loss of STING recruitment, demonstrating a sequential recruitment of cellular PRRs. ChIP analysis further supported this finding as cGAS interaction with HSV-1 genomes was enhanced following UV irradiation, whilst STING demonstrated no interaction at all. Conversely, UV irradiation reduced frequency of recruitment of key PML-NB proteins to vDNA in the nucleus. ChIP analysis demonstrated the interaction between UV irradiated vDNA and PML to be significantly reduced compared to WT HSV-1 genomes. The click labelling was further extended to human Cytomegalovirus (hCMV) to investigate if these immune defences are conserved among Herpesviruses or HSV-1 specific. Collectively, our analysis shows multiple host factors to alter their respective recruitment and/or vDNA binding properties in response to infection following UV irradiation.

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Subjects: Q Science > QR Microbiology > QR355 Virology
Colleges/Schools: College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
Funder's Name: Medical Research Council (MRC)
Supervisor's Name: Boutell, Dr. Chris
Date of Award: 2022
Depositing User: Theses Team
Unique ID: glathesis:2022-82802
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 13 Apr 2022 15:04
Last Modified: 01 Aug 2022 08:43
Thesis DOI: 10.5525/gla.thesis.82802

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