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Uterine epithelial cell release of MIP3, TNF and TGF in response to Escherichia coli, Lactobicillus rhamnosus and pathogen associated molecular patterns (PAMP) and the influence of estradiol on cytokine release.

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Call No.	QP88.4 .C73 2004
Author	Crane-Godreau, Margaret A.
Title	Uterine epithelial cell release of MIP3, TNF and TGF in response to Escherichia coli, Lactobicillus rhamnosus and pathogen associated molecular patterns (PAMP) and the influence of estradiol on cytokine release.
Date	2004
Abstract	The research presented in this thesis tested the hypothesis that uterine epithelial cells (UEC) are selectively responsive to bacteria and pathogen associated molecular patterns (PAMP) and that responses are influenced by estradiol (E 2 ). We developed a model system to define microbially elicited epithelial cell basolateral cytokine release, in which UEC from adult rats were grown to confluence in cell culture inserts and monitored for polarization as determined by transepithelial resistance (TER). Polarized UEC were treated apically with live or heat-killed (HK) Escherichia coli ( E. coli ) or Lactobacillus rhamnosus ( L. rhamnosus ) prior to collection of basolateral media after 24 hr incubation. Co-culture of polarized UEC with live E. coli had no effect on UEC TER. However, in response to live E. coli exposure, UEC release of MIP3α and TNFα increased at a time when release of TGFβ decreased. Incubation of UEC with HK- E. coli stimulated release of MIP3α and TNFα, without affecting TER or TGFβ. While UEC were responsive to E. coli , live or HK L. rhamnosus had no effect on TER or cytokine release. In response to LPS, LTA and Pam 3 Cys, MIP3α and TNFα, release increased. Time course studies of MIP3α and TNFα, production in response to PAMP indicated that MIP3α release peaked 2 to 4 hours after treatment whereas TNFα, release was gradual. To determine the effects of E 2 on MIP3α and TNFα release, UEC were cultured in media with stripped fetal bovine serum (F12K-SS) which contains estrogen concentrations below the level of detection. When UEC cultures were pre-cultured in F12K-SS for 24 hr followed by 24 hr treatment in F12K-SS with estradiol, constitutive release of both MIP3α and TNFα, was inhibited. However when UEC were treated with PAMP, E 2 stimulated MIP3α release at a time when TNFα release was inhibited relative of UEC treated with PAMP alone. The estrogen receptor antagonist ICI 182,780 failed to reverse constitutive-E 2 -inhibitory effects on MIP3α and TNFα. In contrast, treatment with ICI 182,780 reversed the stimulatory effect of E 2 on PAMP-stimulated MIP3α release. Taken together, these findings suggest that UEC responses to bacteria are selective and that estradiol regulates the release of MIP3α and TNFα by UEC.
Description	xi, 145 leaves : ill. 28 cm.
Format	application/pdf
Advisor	Charles R. Wira.
Subject	Epithelial cells. Estradiol -- Physiological effect. Tumor necrosis factor. Transforming growth factors-beta. Cytokines -- Pathophysiology. Escherichia coli. Lactobacillus rhamnosus. Uterus -- Pathophysiology.
College	Dartmouth Medical School -- Department of Physiology.
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D.)--Dartmouth College.
Copyright	All rights reserved. No part of this book may be reproduced in any form by any electronic or mechanical means without permission in writing from the author.
DOI	http://dx.doi.org/10.1349/ddlp.44

Title	Page 1
Format	application/pdf
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D.)--Dartmouth College.
Full Text	Uterine Epithelial Cell Release of MIP3α, TNFα and TGFβ in Response to Escherichia coli, Lactobacillus rhamnosus and Pathogen Associated Molecular Patterns (PAMP) and the Influence of Estradiol on Cytokine Release. A Thesis Submitted to the Faculty in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Physiology by Margaret A. Crane-Godreau DARTMOUTH COLLEGE Hanover, New Hampshire May 21, 2004 Examining Committee: ____________________________ Charles R. Wira, Ph.D. (Chair) ____________________________ Allan U. Munck, Ph.D. ____________________________ Paul M. Guyre, Ph.D. ____________________________ Eugene E. Nattie, M.D. _____________________________ Ronald K. Taylor, Ph.D. ____________________________ Charu Kaushic, Ph.D. _____________________________________ Carol L. Folt, Ph.D., Dean of Graduate Studies

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Uterine epithelial cell release of MIP3, TNF and TGF in response to Escherichia coli, Lactobicillus rhamnosus and pathogen associated molecular patterns (PAMP) and the influence of estradiol on cytokine release.

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