| |
| author |
David Hoyt
| | title |
Investigating the Innate Immune Response to Single-Stranded DNA : A Thesis in Biochemistry and Molecular Biology
| | abstract |
The innate immune system is a crucial part of the immune response to viruses, using
pattern recognition receptors (PRRs) to recognize microbe-associated molecular patterns
(MAMPs) to respond quickly and limit the spread of the virus. Interferon-inducible protein 16
(IFI16) binds to viral double-stranded DNA, which triggers the production of inflammatory
cytokines and Type 1 interferons like Interferon-Beta (IFN-B). Previous studies have shown that
there does not appear to be the production of IFN-B 6 hours after stimulation using ssVac70,
while there is a response to dsVac70. However, further investigation revealed to possibility of a
delayed response at 24 hours through the transcription of ISG56. Based on the observed delayed
responses, I hypothesized that there is an innate response to ssVac70, but that response is smaller
and different in downstream products when compared to the response dsVac70. I also
hypothesized that the response to ssVac70 is reliant on the generation of a dsDNA intermediate,
and the use of aphidicolin, a DNA polymerase inhibitor, will eliminate the response. The innate
immune response to ssVac70 was investigated by transfecting mature THP-1 cells with ssVac70
and measuring four innate immune genes: IFN-B, ISG56, OASL, and IL-6. The results of all four
genes shows that ssVac70 induces an innate immune response particularly at 6 hours, but that
response is smaller than the response to dsVac70. Aphidicolin treatment caused a loss of the
response at 6 hours, suggesting that the dsDNA intermediate is required for this response.
Differences between the 6 and 24 hour timepoints suggest that there may be a different
mechanism operating at each of the timepoints. Ultimately, these findings support further
research in the response to ssDNA across a variety of different applications.
| | school |
The College of Liberal Arts, Drew University
| | degree |
B.S. (2024)
|
| advisor |
Brianne Barker
|
| full text | DHoyt.pdf - requires Drew uLogin |
| |
