Access to nectin favors herpes simplex virus infection at the apical surface of polarized human epithelial cells

Benjamin T. Galen, Natalia V. Cheshenko, Ana Tuyama, Bharat Ramratnam, Betsy Herold

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Viral entry may preferentially occur at the apical or the basolateral surfaces of polarized cells, and differences may impact pathogenesis, preventative strategies, and successful implementation of viral vectors for gene therapy. The objective of these studies was to examine the polarity of herpes simplex virus (HSV) entry using several different human epithelial cell lines. Human uterine (ECC-1), colonic (CaCo-2), and retinal pigment (ARPE-19) epithelial cells were grown on collagen-coated inserts, and the polarity was monitored by measuring the transepithelial cell resistance. Controls were CaSki cells, a human cervical cell line that does not polarize in vitro. The polarized cells, but not CaSki cells, were 16- to 50-fold more susceptible to HSV infection at the apical surface than at the basolateral surface. Disruption of the tight junctions by treatment with EGTA overcame the restriction on basolateral infection but had no impact on apical infection. No differences in binding at the two surfaces were observed. Confocal microscopy demonstrated that nectin-1, the major coreceptor for HSV entry, sorted preferentially to the apical surface, overlapping with adherens and tight junction proteins. Transfection with small interfering RNA specific for nectin-1 resulted in a significant reduction in susceptibility to HSV at the apical surface but had little impact on basolateral infection. Infection from the apical but not the basolateral surface triggered focal adhesion kinase phosphorylation and led to nuclear transport of viral capsids and viral gene expression. These studies indicate that access to nectin-1 contributes to preferential apical infection of these human epithelial cells by HSV.

Original languageEnglish (US)
Pages (from-to)12209-12218
Number of pages10
JournalJournal of Virology
Volume80
Issue number24
DOIs
StatePublished - Dec 2006
Externally publishedYes

Fingerprint

herpes simplex
Virus Diseases
Simplexvirus
epithelial cells
Epithelial Cells
viruses
Infection
Virus Internalization
infection
Viral Genes
tight junctions
cells
Tight Junction Proteins
non-specific protein-tyrosine kinase
Adherens Junctions
Cell Line
Focal Adhesion Protein-Tyrosine Kinases
cell lines
Retinal Pigments
Cell Nucleus Active Transport

ASJC Scopus subject areas

  • Immunology

Cite this

Access to nectin favors herpes simplex virus infection at the apical surface of polarized human epithelial cells. / Galen, Benjamin T.; Cheshenko, Natalia V.; Tuyama, Ana; Ramratnam, Bharat; Herold, Betsy.

In: Journal of Virology, Vol. 80, No. 24, 12.2006, p. 12209-12218.

Research output: Contribution to journalArticle

@article{db4058b298574798b3077f24fdc4e53a,
title = "Access to nectin favors herpes simplex virus infection at the apical surface of polarized human epithelial cells",
abstract = "Viral entry may preferentially occur at the apical or the basolateral surfaces of polarized cells, and differences may impact pathogenesis, preventative strategies, and successful implementation of viral vectors for gene therapy. The objective of these studies was to examine the polarity of herpes simplex virus (HSV) entry using several different human epithelial cell lines. Human uterine (ECC-1), colonic (CaCo-2), and retinal pigment (ARPE-19) epithelial cells were grown on collagen-coated inserts, and the polarity was monitored by measuring the transepithelial cell resistance. Controls were CaSki cells, a human cervical cell line that does not polarize in vitro. The polarized cells, but not CaSki cells, were 16- to 50-fold more susceptible to HSV infection at the apical surface than at the basolateral surface. Disruption of the tight junctions by treatment with EGTA overcame the restriction on basolateral infection but had no impact on apical infection. No differences in binding at the two surfaces were observed. Confocal microscopy demonstrated that nectin-1, the major coreceptor for HSV entry, sorted preferentially to the apical surface, overlapping with adherens and tight junction proteins. Transfection with small interfering RNA specific for nectin-1 resulted in a significant reduction in susceptibility to HSV at the apical surface but had little impact on basolateral infection. Infection from the apical but not the basolateral surface triggered focal adhesion kinase phosphorylation and led to nuclear transport of viral capsids and viral gene expression. These studies indicate that access to nectin-1 contributes to preferential apical infection of these human epithelial cells by HSV.",
author = "Galen, {Benjamin T.} and Cheshenko, {Natalia V.} and Ana Tuyama and Bharat Ramratnam and Betsy Herold",
year = "2006",
month = "12",
doi = "10.1128/JVI.01503-06",
language = "English (US)",
volume = "80",
pages = "12209--12218",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "24",

}

TY - JOUR

T1 - Access to nectin favors herpes simplex virus infection at the apical surface of polarized human epithelial cells

AU - Galen, Benjamin T.

AU - Cheshenko, Natalia V.

AU - Tuyama, Ana

AU - Ramratnam, Bharat

AU - Herold, Betsy

PY - 2006/12

Y1 - 2006/12

N2 - Viral entry may preferentially occur at the apical or the basolateral surfaces of polarized cells, and differences may impact pathogenesis, preventative strategies, and successful implementation of viral vectors for gene therapy. The objective of these studies was to examine the polarity of herpes simplex virus (HSV) entry using several different human epithelial cell lines. Human uterine (ECC-1), colonic (CaCo-2), and retinal pigment (ARPE-19) epithelial cells were grown on collagen-coated inserts, and the polarity was monitored by measuring the transepithelial cell resistance. Controls were CaSki cells, a human cervical cell line that does not polarize in vitro. The polarized cells, but not CaSki cells, were 16- to 50-fold more susceptible to HSV infection at the apical surface than at the basolateral surface. Disruption of the tight junctions by treatment with EGTA overcame the restriction on basolateral infection but had no impact on apical infection. No differences in binding at the two surfaces were observed. Confocal microscopy demonstrated that nectin-1, the major coreceptor for HSV entry, sorted preferentially to the apical surface, overlapping with adherens and tight junction proteins. Transfection with small interfering RNA specific for nectin-1 resulted in a significant reduction in susceptibility to HSV at the apical surface but had little impact on basolateral infection. Infection from the apical but not the basolateral surface triggered focal adhesion kinase phosphorylation and led to nuclear transport of viral capsids and viral gene expression. These studies indicate that access to nectin-1 contributes to preferential apical infection of these human epithelial cells by HSV.

AB - Viral entry may preferentially occur at the apical or the basolateral surfaces of polarized cells, and differences may impact pathogenesis, preventative strategies, and successful implementation of viral vectors for gene therapy. The objective of these studies was to examine the polarity of herpes simplex virus (HSV) entry using several different human epithelial cell lines. Human uterine (ECC-1), colonic (CaCo-2), and retinal pigment (ARPE-19) epithelial cells were grown on collagen-coated inserts, and the polarity was monitored by measuring the transepithelial cell resistance. Controls were CaSki cells, a human cervical cell line that does not polarize in vitro. The polarized cells, but not CaSki cells, were 16- to 50-fold more susceptible to HSV infection at the apical surface than at the basolateral surface. Disruption of the tight junctions by treatment with EGTA overcame the restriction on basolateral infection but had no impact on apical infection. No differences in binding at the two surfaces were observed. Confocal microscopy demonstrated that nectin-1, the major coreceptor for HSV entry, sorted preferentially to the apical surface, overlapping with adherens and tight junction proteins. Transfection with small interfering RNA specific for nectin-1 resulted in a significant reduction in susceptibility to HSV at the apical surface but had little impact on basolateral infection. Infection from the apical but not the basolateral surface triggered focal adhesion kinase phosphorylation and led to nuclear transport of viral capsids and viral gene expression. These studies indicate that access to nectin-1 contributes to preferential apical infection of these human epithelial cells by HSV.

UR - http://www.scopus.com/inward/record.url?scp=33845439431&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33845439431&partnerID=8YFLogxK

U2 - 10.1128/JVI.01503-06

DO - 10.1128/JVI.01503-06

M3 - Article

VL - 80

SP - 12209

EP - 12218

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 24

ER -