Infection of Polarized Primary Epithelial Cells from Rat Uterus with Chlamydia trachomatis: Cell–Cell Interaction and Cytokine Secretion
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abstract
PROBLEM: The objective of this study was to examine the susceptibility of rat uterine epithelial cells (UEC) to infection with Chlamydia
trachomatis and to study the epithelial–stromal interactions
following infection.
METHOD OF STUDY: UEC were isolated from adult rats and grown in culture.
Polarized, confluent monolayers of UEC were infected with 106
IFU/well C. trachomatis (MoPn). In order to confirm infection,
MoPn was labeled with a fluorescent tracking dye, PKH‐26, and then used
in epithelial cell infections. Transepithelial resistances were measured
prior to and following infection to test the effect of Chlamydia
on the integrity of the epithelial monolayers. In other experiments,
polarized epithelial cultures were infected in the presence and absence
of stromal cells. Media was collected from the apical and basolateral
compartments of the cultures before and after infection and analyzed for
cytokines IL‐1α and TNF‐α.
RESULTS: Epithelial cell cultures infected with PKH‐26 labeled MoPn were
examined 4–5 days later. Bacterial inclusions were detected inside
epithelial cells indicating infection had occurred. Co‐localization of
PKH‐26 labeled bacteria with FITC‐labelled anti‐Chlamydia antibody
on the epithelial cells confirmed infection. No changes were found in
resistance across the monolayers of epithelial cells in the presence or
absence of infection. ELISA results indicate that UEC secrete IL‐1α
constitutively in vitro. Stromal cells secrete very little
IL‐1α. When stromal cells were co‐incubated with epithelial cells
there was a decrease in the amount of IL‐1α secreted by epithelial
cells 48 hr post‐infection. On the other hand, maximum TNF‐α was
found in stromal cells, both with and without infection. Epithelial
cells, in these studies made very little TNF‐α.
CONCLUSIONS: These results show that primary rat epithelial cells can be
infected with Chlamydia in vitro. Epithelial and stromal cells
from uteri of adult rats make IL‐1α and TNF‐αin vitro
both prior to and following infection with Chlamydia. This system can be used to analyze the role played by epithelial–stromal interactions in providing protection on this mucosal surface.
