Mathematical Model of Viral Kinetics In Vitro Estimates the Number of E2-CD81 Complexes Necessary for Hepatitis C Virus Entry

Interaction between the hepatitis C virus (HCV) envelope protein E2 and the host receptor CD81 is essential for HCV entry into target cells. The number of E2-CD81 complexes necessary for HCV entry has remained difficult to estimate experimentally. Using the recently developed cell culture systems that allow persistent HCV infection in vitro, the dependence of HCV entry and kinetics on CD81 expression has been measured. We reasoned that analysis of the latter experiments using a mathematical model of viral kinetics may yield estimates of the number of E2-CD81 complexes necessary for HCV entry. Here, we constructed a mathematical model of HCV viral kinetics in vitro, in which we accounted explicitly for the dependence of HCV entry on CD81 expression. Model predictions of viral kinetics are in quantitative agreement with experimental observations. Specifically, our model predicts triphasic viral kinetics in vitro, where the first phase is characterized by cell proliferation, the second by the infection of susceptible cells and the third by the growth of cells refractory to infection. By fitting model predictions to the above data, we were able to estimate the threshold number of E2-CD81 complexes necessary for HCV entry into human hepatoma-derived cells. We found that depending on the E2-CD81 binding affinity, between 1 and 13 E2-CD81 complexes are necessary for HCV entry. With this estimate, our model captured data from independent experiments that employed different HCV clones and cells with distinct CD81 expression levels, indicating that the estimate is robust. Our study thus quantifies the molecular requirements of HCV entry and suggests guidelines for intervention strategies that target the E2-CD81 interaction. Further, our model presents a framework for quantitative analyses of cell culture studies now extensively employed to investigate HCV infection. Author Summary: The interaction between the hepatitis C virus (HCV) envelope protein E2 and the host cell surface receptor CD81 is critical for HCV entry into hepatocytes and presents a promising drug and vaccine target. Yet, the number of E2-CD81 complexes that must be formed between a virus and a target cell to enable viral entry remains unknown. Direct observation of the E2-CD81 complexes preceding viral entry has not been possible. We constructed a mathematical model of HCV viral kinetics in vitro and using it to analyze data from recent cell culture studies obtained estimates of the threshold number of E2-CD81 complexes necessary for HCV entry. We found that depending on the E2-CD81 binding affinity, between 1 and 13 complexes are necessary for HCV entry into human hepatoma-derived cells. Our study thus presents new, quantitative insights into the molecular requirements of HCV entry, which may serve as a guideline for intervention strategies targeting the E2-CD81 interaction. Further, our study shows that HCV viral kinetics in vitro can be described using a mathematical model, thus facilitating quantitative analyses of the wealth of data now emanating from cell culture studies of HCV infection.