Date of Graduation

Spring 2024

Document Type

Thesis

Degree Name

Master of Science in Biology

First Advisor

Sangman Kim

Second Advisor

Naupaka Zimmerman

Third Advisor

Christina Tzagarakis-Foster

Abstract

Lyme disease is the most common vector-borne disease in the US and will experience an uptick as the insect host, the ixodid tick, gains habitat with climate change. The causative agent of Lyme disease, Borrelia burgdorferi, is recognized by Toll-like receptor 2 (TLR2), which initiates the innate immune response. Here, I used ELISA and SEAP detection assays to determine that knocking out TLR2 in THP-1 macrophages reduces the amount of NFkB activation, as well as IL-10 and IL-1β secretion. Then, I used fluorescence microscopy and Incucyte assays to quantify the amount of phagocytosis performed by wild-type and knock-out THP-1 cells. I found that despite the changes in signaling, knock-out of TLR2 does not affect the extent of phagocytosis. These results support previous studies in mice that found that TLR2 knock-out impairs cytokine response but not phagocytic responses. Finally, I used a SEAP detection assay to measure the change in NFkB activation in HEK293T cells transfected with TLR2 mutants and stimulated with Borrelia. I found that 2 mutations, T411I and R753Q impair the NFkB activation response. Further study of these mutations may help to develop vaccines against Lyme disease by increasing understanding of the heterogeneity in immune responses to Borrelia.

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