TRUSTEES OF BOSTON UNIVERSITY
These supplemental funds will be used to create a new position for a graduate student who is being added to this project. This student worked part-time on the project over the past year while being supported by a scholarship, and discovered that junctional adhesion molecules (JAMs) may play a role in macrophage attachment and infiltration of cervicovaginal epithelia. Her research fits within the scope of this project, yet adds a new dimension that will significantly enhance and accelerate the tempo of our research. Our original research plan did not include studies on intracellular junctional adhesion molecule (JAM) proteins or epithelial permeability, but we realize now, as a result of her preliminary research, that they are highly relevant to our research on cell-associated HIV transmission. It is likely that HIV-infected macrophages use JAM proteins to attach to and infiltrate vaginal and cervical epithelia. She plans to characterize the expression and function of these proteins in normal human vaginal and endocervical tissues and in the MatTekEpiVaginal(TM) model, and to study the effects of reproductive hormones, inflammation and seminal plasma. She will use immunohistology to localize, and RT_PCR to quantify the expression of JAM-A and JAM-C in tissue explants and the MatTekEpiVaginal model, and will use blocking antibodies and peptides in a newly developed quantitative confocal microscopy adhesion/infiltration assay to determine whether JAM proteins play a role in macrophage attachment and infiltration of vaginal tissue explants and EpiVaginal organotypic cultures, and whether they can be used to block of cell-associated HIV transmission. This research could provide important information on the molecular mechanisms of macrophage-epithelial interactions and suggest new strategies for blocking HIV transmission.