Lyme disease, caused by the Gram-negative bacterium Borrelia burgdorferi (Bb), is recognized as one of the most common vector-borne infectious diseases. It has raised public health concerns in many parts of the US and across the globe. Although there are treatments available for early-stage Lyme disease such as amoxicillin, doxycycline, and other common oral antibiotics, we aim to develop a vaccine that offers a preventive strategy beyond the current reliance on antibiotics. Vaccines provide broader protection against Lyme disease across the population. A mucosal vaccine is favored since it induces immunity against Bb infection and elicits both local mucosal immunity and systemic immunity. In addition, mucosal vaccines can be administered non-invasively, making them more acceptable to the public and improving access to vaccination. Outer Membrane Vesicles (OMVs), nanoparticles (NPs) that bud off from the outer membrane of Gram-negative bacteria, are considered potential vaccine platforms due to their unique immunogenic properties. These NPs are non-replicating and natural pathogen mimics, as they carry a variety of bacterial components and pathogen-associated molecular patterns, including proteins, lipopolysaccharides, and genetic materials. Therefore, OMVs can be recognized by pattern recognition receptors, which are expressed on immune cells. In this study, we aim to develop a mucosal vaccine utilizing OMVs as a platform. From that, we want to ascertain whether the vaccine particle size plays a role in the generation of immunity against the pathogens. As OMVs are around 100 nm in diameter and bacteria are around 1 µm, conjugation of OMVs is performed to subsequently crosslink OMVs together, forming microscale-sized particles. The resultant microparticles will be examined for their ability to interact with the immune system similarly to bacteria. So far, the amount of available reactive amine groups present on the OMVs has been obtained. Using this value, the crosslinking ratios between the OMVs and polyethylene glycol (PEG)-based cross-linkers are currently being evaluated. We expect that cross-linked OMVs of desired size (~1 um) will be successfully produced. Following this, we will observe the behavior of the particles over time at various temperatures (4oC, 20oC, and 37oC) to detect possible degradation. Controlled degradation in human saliva is necessary for induction of an appropriate systemic and mucosal immune response in future studies.
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