Phosphorylcholine (PC) has been reported to help PC-bearing Aggregatibacter actinomycetemcomitans to gain access to the general circulation via interaction with the platelet-activating factor receptor (PAFR) on endothelial cells. However, the underlying mechanism remains unclear. Here, we found that inhibition of clathrin and β-arrestins blocked bacterial PC binding to PAFR which significantly attenuated A. actinomycetemcomitans invasion of human vascular endothelial cells (HUVEC). These results demonstrated the involvement of clathrin and β-arrestins in PC-bearing A. actinomycetemcomitans invasion of HUVEC.