P. gingivalis is a Gram-negative anaerobe that is considered one of the etiologic agents of certain types of periodontal disease. In addition, there is significant evidence that P. gingivalis may play a role in the progression of cardiovascular diseases (CVD), especially atherosclerosis. We have previously reported that P. gingivalis can invade primary human cardiovascular endothelial cells (HCAEC), the cells where the initial events of atherosclerosis take place. We have determined that once inside the cells, P. gingivalis usurps the autophagic pathway of the HCAEC and uses autophagic vesicles as its privileged niche where it survives and replicates. The purpose of the current application is to more completely define the communication and interactions at the molecular level between the bacteria and its host. Based on our previous studies, our hypothesis is that P. gingivalis secretes or expresses proteins that are required for adhesion/internalization, trafficking and persistence in HCAEC. In Aim 1, we will identify those proteins that are essential for P. gingivalis adhesion/internalization, trafficking and persistence in HCAEC by mutant analysis. In Aim 2, we will more fully characterize the proteins verified to be adhesins or essential for internalization and determine if internalization proceeds via lipid rafts. In Aim 3, we will study bacterial trafficking within HCAEC by identifying and characterizing those proteins essential for activating autophagy and trafficking the bacterium away from the lysosomes and to the replicating vacuoles. In Aim 4, we will examine bacterial persistence by identifying and characterizing those proteins that promote survival of HCAEC. Finally, in Specific Aim 5, we will characterize the expression profiles in HCAEC infected with wild type and mutant strains of P. gingivalis. This comprehensive study will provide us essential information on the molecular events required for invasion of endothelial cells by this intracellular pathogen. [unreadable] [unreadable]