The effect of cystic fibrosis environment on the bactericidal and phagocytic abilities of macrophages Public

Abstract

Abstract
Alveolar cystic fibrosis (CF) macrophages have decreased innate bactericidal abilities. Compared to non-CF macrophages in the airway, CF macrophages are surrounded with an overabundance of cytokines and a thick layer of dehydrated mucous. To test the effect of the CF environment on the bactericidal and phagocytic ability of the macrophage, we conducted in vitro bactericidal assays. We infected mouse leukaemic monocyte macrophage cell line (RAW 264.7) macrophages with Pseudomonas aeruginosa (PA01) after conditioning the cells with CF plasma, non-CF plasma, tumor necrosis factor α (TNFα), interleukin-8 (IL-8), or CF sputum. Additionally, we performed an in vivo bactericidal test in which we infected CF and wild-type mice with PA01 and measured bacterial proliferation in the bronchoalveolar fluid (BAF) and lungs. Furthermore, we tested the respiratory burst of primary CF and non-CF monocytes, measured the level of pro-inflammatory cytokines TNFα and IL-6 released by primary CF monocytes after infection with PA01, and observed the internalization of PA01 by CF macrophages. Although the CF monocytes appeared to be able to internalize bacteria, produce reactive oxygen species, and signal other cells via cytokines or chemokines, they exhibited decreased bactericidal abilities when tested in vivo or with TNFα or IL-8. Thus, we conclude that the inflammatory nature of the CF environment inhibits the bactericidal abilities of macrophages.

Table of Contents

Table of Contents

Introduction……………………………………………………...1
Materials and Methods…………………………………........5

Figure 1 …………………………………………………………………………….…8
Figure 2 …………………………………………………………………………….…9
Figure 3 ………………………………………………………………………………10
Figure 4…………………………………………………………………………….…12

Results……………………………………………………………..13
Figure 5……………………………………………………………………………….13
Figure 6……………………………………………………………………………….14
Figure 7……………………………………………………………………………….15
Figure 8……………………………………………………………………………….16
Figure 9……………………………………………………………………………….17
Figure 10………………………………………………………………………...….17
Figure 11..…………………………………………………………………….…….19
Figure 12..………………………………………………………………….……….20

Discussion……………………………………………...…..……..21
Future Directions………………………………….…………….23
References.………………………………………..…………..….25

About this Honors Thesis

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School	Emory College
Department	Biology
Degree	BS
Submission	Honors Thesis
Language	English
Research Field	Biology, General Biology, Microbiology
Mot-clé	cystic fibrosis, CF, macrophage, immunology, microbiology, cytokines, sputum, bactericidal, phagocytic, reactive oxygen species, ELISA
Committee Chair / Thesis Advisor	McCarty, Nael, Emory University
Committee Members	Antia, Rustom, Emory University Passalacqua, Karla D, Emory University Zughaier, Susu M, Emory University

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