The mucosal surface is the site of entry of >90% of common infectious pathogens in humans. However, studies of virus entry and infection are largely limited to non-polarized cells and little is known regarding the mechanisms used by viral pathogens to infect polarized cell monolayers. Enteroviruses are ideal pathogens for the study of polarized cell-virus interactions as they have evolved highly efficient strategies to bypass both epithelial and endothelial barriers for infection. Our previous studies have established that coxsackievirus B (CVB) and poliovirus (PV) enter polarized cells by endocytic mechanisms that require activation of specific intracellular signaling molecules that drive actin cytoskeleton reorganization, junctional complex modulation, and eventual virus endocytosis. Our preliminary studies suggest that CVB endocytosis into polarized intestinal epithelia is distinct from that in polarized BBB endothelia and that the signaling molecules required to facilitate virus entry are disparate between these cell types. Based on these observations, we hypothesize that enterovirus entry into polarized epithelial and endothelial cells is mediated by cell-type-specific intracellular signals and occur through different endocytic mechanisms. In this proposal, we have designed experiments to more clearly define the cellular pathways hijacked by CVB to gain entry into polarized human brain microvascular endothelial cells (HBMEC), a model of the blood-brain barrier. Specifically, we will (1) define the endocytic pathway used by CVB to gain entry into HBMEC (Specific Aim 1), (2) characterize the role of intracellular kinases in facilitating CVB entry into HBMEC (Specific Aim 2), and (3) compare the intracellular signaling molecules required for enterovirus infection in a variety of cell types (Specific Aim 3). Given the diverse pathological complications associated with enterovirus infections, these studies will provide insights into how enteroviruses have evolved to infect polarized cells, and will serve as a model for how other viral pathogens circumvent the barriers presented by polarized cell monolayers. Furthermore, as there are currently no effective therapeutics to treat enterovirus infections, these studies may lead to the design of cell-type-specific anti-viral targets.

Public Health Relevance

Enteroviruses (which include coxsackievirus, poliovirus, and echovirus) account for as many as 15 million symptomatic infections in the United States each year. There are currently no effective therapeutics to combat enteroviral infections. The studies in this proposal will identify cellular genes required for mediating enterovirus infection and may lead to the design of cell-type-specific antiviral targets.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
Virology - A Study Section (VIRA)
Program Officer
Park, Eun-Chung
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Pittsburgh
Schools of Medicine
United States
Zip Code
Morosky, Stefanie; Lennemann, Nicholas J; Coyne, Carolyn B (2016) BPIFB6 Regulates Secretory Pathway Trafficking and Enterovirus Replication. J Virol 90:5098-107
Bayer, Avraham; Lennemann, Nicholas J; Ouyang, Yingshi et al. (2016) Type III Interferons Produced by Human Placental Trophoblasts Confer Protection against Zika Virus Infection. Cell Host Microbe 19:705-12
McConkey, Cameron A; Delorme-Axford, Elizabeth; Nickerson, Cheryl A et al. (2016) A three-dimensional culture system recapitulates placental syncytiotrophoblast development and microbial resistance. Sci Adv 2:e1501462
Ouyang, Yingshi; Bayer, Avraham; Chu, Tianjiao et al. (2016) Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity. Placenta 47:86-95
Drummond, Coyne G; Nickerson, Cheryl A; Coyne, Carolyn B (2016) A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells. mSphere 1:
Harris, Katharine G; Coyne, Carolyn B (2015) Unc93b Induces Apoptotic Cell Death and Is Cleaved by Host and Enteroviral Proteases. PLoS One 10:e0141383
Bayer, Avraham; Delorme-Axford, Elizabeth; Sleigher, Christie et al. (2015) Human trophoblasts confer resistance to viruses implicated in perinatal infection. Am J Obstet Gynecol 212:71.e1-8
Harris, Katharine G; Morosky, Stefanie A; Drummond, Coyne G et al. (2015) RIP3 Regulates Autophagy and Promotes Coxsackievirus B3 Infection of Intestinal Epithelial Cells. Cell Host Microbe 18:221-32
Sadovsky, Yoel; Mouillet, Jean-Francois; Ouyang, Yingshi et al. (2015) The Function of TrophomiRs and Other MicroRNAs in the Human Placenta. Cold Spring Harb Perspect Med 5:a023036
Mouillet, Jean-François; Ouyang, Yingshi; Coyne, Carolyn B et al. (2015) MicroRNAs in placental health and disease. Am J Obstet Gynecol 213:S163-72

Showing the most recent 10 out of 29 publications