Using 3-D Primary Mucosal Cultures to Study Antibody Function at Mucosal Barrier

There is currently a great need to develop a vaccine for HIV. The focus on a vaccine that elicits broadly neutralizing antibodies is noble and obvious, but has yet to show promise as a vaccine approach. To achieve this goal we must have a better understanding of how antibodies function at intact mucosal barriers where the virus is transmitted. In this pilot project, we seek to adapt a primary mucosal tissue culture model developed in Innsbruck that accurately recapitulates native ciliated mucosal of barriers. Importantly, this system produces mucus and has an intact glycocalyx, which is like the native system. We will adapt this system to study HIV utilizing fluorescently tagged antibody and virus developed in Chicago. The expertise of the labs in Austria and the US are highly complementary and should allow this culture system to be used to study HIV interaction with native Mucosal barriers. To this end, we will define this system for the expression of mucins, evaluate the interaction of antibodies with this barrier, and then determine how HIV diffusion and interaction with the Mucosal barrier is modified by the presence of the antibodies. This pilot project has a great potential to be highly significant and impactful. It will provide novel insights into how native mucosal barriers function to inhibit virus penetration, and provide important insights into how antibodies can be used to reinforce this barrier. It also sets the stage for the development of other systems that more accurately recapitulates environments of the upper female reproductive tract, and colorectal system. This pilot project will also set the stage for larger more comprehensive projects that allow this system to be dissected in detail, and adapt this system to be utilized for a variety of questions relating to HIV prevention.