ORCID

https://orcid.org/0009-0007-1529-7934

Date of Award

Summer 2026

Language

English

Embargo Period

7-20-2026

Document Type

Master's Thesis

Degree Name

Master of Science (MS)

College/School/Department

Department of Biomedical Sciences

Program

Biomedical Sciences

First Advisor

Nicholas Mantis

Committee Members

April Davis, Jennifer Yates

Keywords

Borrelia burgdorferi, Lyme Disease, Infection and Immunity, Immunofluorescence, Confocal Microscopy, Mouse Model

Subject Categories

Immunology and Infectious Disease | Public Health

Abstract

The skin is an immunological organ that acts as the hosts’ first line of defense against infectious agents. Infectious disease vectors like ticks breach this barrier by feeding on hosts, transmitting pathogens like Borrelia burgdorferi in the process. Uncleared infection with B. burgdorferi causes Lyme disease, the most prevalent vector-borne disease in the United States. Previous research has explained many elements of Lyme disease pathogenesis and immunity including the dissemination of the spirochete, innate immune cells performing phagocytosis, the production of antibodies by the humoral response, and the impact of prophylactic treatment with monoclonal antibodies. A remaining gap in the field is the ability to visualize pathogen-host immune response interactions within in situ contexts.

In this thesis, I have developed and implemented a protocol utilizing a mouse model, skin tissue cryosectioning, immunofluorescence, and confocal microscopy to interrogate pathogen and host immune response dynamics during early B. burgdorferi infection in the skin. After optimization of the experimental procedure, I applied it to preliminarily study B. burgdorferi presence under infection conditions and explored different mechanisms contributing to infection outcome including of innate-, vaccine-, and adaptive-based immunity.

Using these techniques, I observed an increase in B. burgdorferi numbers 5 days post inoculation, with apparent dissemination away from the site of inoculation 7 days post inoculation. Also, innate immune cells and pro-inflammatory cytokine and chemokine presence matched with B. burgdorferi presence, indicating correlation between these elements of infection. In addition, I found that bacterial burden was increased in mice lacking the TLR adaptor protein, MyD88. Interestingly, innate immune cell recruitment did not appear to be impaired in the knockout mice, but instead MyD88 may impact innate immune cell morphology and phagocytosis of B. burgdorferi. Next, prophylactic treatment with anti-OspA monoclonal antibody, LA-2, demonstrated passive protection in in situ fluorescent images as demonstrated by decreased B. burgdorferi abundance, earlier innate immune cell response, and lower pro-inflammatory cytokine and chemokine presence across timepoints compared to infection groups. Lastly, a panel of recombinant monoclonal antibodies derived from the Erythema migrans skin lesion of Lyme disease patients were characterized based on observed reactivity to B. burgdorferi or host cells to predict protective abilities or autoimmune risk.

The developed protocol can be employed to validate published literature, interpret and corroborate findings produced in ongoing projects, and explore unanswered questions remaining in the field of Lyme disease and skin immunology research. This protocol can be used to investigate multiple mechanisms playing a role in infection outcome, including spirochete abundance and spread, phagocytosis performed by innate immune cells, and protection conferred by monoclonal antibodies produced following exposure to B. burgdorferi antigens. These techniques can be leveraged to ultimately contribute diagnosis, treatment, and prevention of Lyme disease, as well as other vector-borne diseases transmitted through the skin.

License

Creative Commons Attribution 4.0 International License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Download

Share

COinS