Comparison of cytokine gene expression in bovine tissues with acute or chronic inflammation

103 Pages·2016·2.88 MB·English

Iowa State University Capstones, Theses and
Retrospective Theses and Dissertations
Dissertations
1998
Comparison of cytokine gene expression in bovine
tissues with acute or chronic inflammation
Haa-Yung Lee
Iowa State University
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Recommended Citation
Lee, Haa-Yung, "Comparison of cytokine gene expression in bovine tissues with acute or chronic inflammation " (1998).Retrospective
Theses and Dissertations. 11942.
https://lib.dr.iastate.edu/rtd/11942
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Comparison of cytokine gene expression in bovine tissues with
acute or chronic inflammation
by
Haa-Yung Lee
A dissertation submitted to the graduate faculty
in partial fulfillment of the requirements for the degree of
DOCTOR OF PHILOSOPHY
Major: Immunobiology
Major Professors: Marcus E. Kehrli, Jr. and Michael J. Wannemuehler
Iowa State University
Ames, Iowa
1998
DMI Number: 9911617
UMI Microform 9911617
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Graduate College
Iowa State University
This is to certify that the Doctoral dissertation of
Haa-Yung Lee
has met the dissertation requirements of Iowa State University
Signature was redacted for privacy.
Co-major Prof^sor
Signature was redacted for privacy.
Co-n>aior Protessor
Signature was redacted for privacy.
Tor ^he^ajor Program
Signature was redacted for privacy.
or the Graduate Colleae
iii
Dedication
To my late father, Chul-Soon Lee, in heaven
To my devoted mother, Rok-Nye Tae
To my beloved wife, Sung-Ok Han
To my lovely daughters, Sara Sang-Hwa and Rachael Hae-Ji
IV
TABLE OF CONTENTS
ABSTRACT
GENERAL INTRODUCTION 1
Dissertation Organization 1
Literature Review 1
Infection and Immune Responses I
Introduction 1
Gram-negative bacterial infection and acute phase cytokines 2
Glucocorticoids in immune response 4
Acute phase cytokines in Disease 5
Tumor necrosis factor (TNF) 5
TNF-receptors (TNFR) 7
Interleukin-1 (IL-1) 8
IL-1 receptors (IL-IR) 9
InterIeukin-6 (IL-6) 11
EL-6 receptor (IL-6R) 12
Interferon-y (IFN-y) 12
Cytokines in Bovine Diseases 13
References 15
EXPRESSION OF ACUTE PHASE CYTOKINE GENES IN BOVINE MAMMARY
TISSUES DURING EXPERLVIENTALLY INDUCED ESCHERICHIA. COLI
MASITIS 30
Abstract 30
Introduction 31
Materials and Methods 31
Animals and Escherichia culi inoculation 31
Analysis of milk samples 32
Probe synthesis for in situ hybridization 32
In situ hybridization 33
Data analysis 34
Results 34
Discussion 35
References 37
V
INFLUENCE OF p,-INTEGRIN ADHESION MOLECULE EXPRESSION AND
PULMONARY INFECTION WITH PASTEURELLA HAEMOLYTICA ON
CYTOKINE GENE EXPRESSION IN CATTLE 49
Abstract 49
Introduction 50
Materials and Methods 51
Animals and Pasteiirelki haemolytica inoculation 51
Probe synthesis for in situ hybridization 51
In situ hybridization 52
Macrophage staining 53
Neutrophil staining 53
Data analysis 53
Results 54
Discussion 56
References 57
CYTOKINE GENE EXPRESSION IN ILEAL TISSUES OF CATTLE INFECTED
WITH MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS 69
Abstract 69
Introduction 69
Materials and Methods 71
Tissue preparation 71
Probe synthesis for in situ hybridization 71
In situ hybridization 72
Macrophage staining 72
Neutrophil staining 73
Data analysis 73
Results 74
Discussion 74
References 77
GENERAL CONCLUSIONS 86
References 88
ACKNOWLEDGEMENTS 92
VI
ABSTRACT
Cytokines are small molecular weight glycoproteins secreted by a v;iriety ot" cells. These
proteins have diverse activities and serve as signal mediators in immune reactions. Among the
stimulators of cytokine induction, bacteria and their products are well-known. However, the
regulatory mechanisms of cytokine expression associated with bacterial diseases are
complicated and not completely elucidated.
In the first study, mammary parenchymal tissues taken from Holstein cows at 6, 12. and
24 hours post-challenge with Escherichia coli were used as an acute bacterial infection model.
In the study, we determined that IL- la and TNF-a were the predominant cytokines expressed
in mammary parenchymal tissues during the early phase of E. cc//-induced mastitis. In
contrast to these cytokines, expression of mRNA for IL-ip. lL-6. and IFN'-yin
£. c6>//-infected mammary gland were low at the time points examined. The second study was
designed to investigate the effects of CD 18, an adhesion molecule, on cytokine gene
expression in bovine pulmonary tissues after respiratory infection with Pasteiirella haemolytica.
This study suggested that CD 18 may contribute to mRNA expression of IFN-yand TNF-a in
the lung of P. haemolytica-in'lccx.Qd. cattle at 4 hours post-challenge. Based on histological
evidence, this observation correlated with the infiltration of neutrophils into the lung tissue.
For the third study, ileal tissues from cattle chronically infected with Mycobacterium avium
subsp. parauiberculosis were examined for the chronic expression of cytokine-specific mRNA.
Ileal tissues collected from cattle with paratuberculosis expressed significantly (P<0.05) more
IL-la. IL-1(3, IL-6 and IFN-ymRNA than paratuberculosis negative cattle. The expression
level of TNF-a, however, was not different between M. avium subsp. parataberculosis-
infected and non-infected animals. Histologically, the ileal tissues from paratuberculosis
positive cows harbored 3 times more macrophages than control cattle.
In combination, these studies suggest that the patterns of cytokine expression and the
predominant subset of leukocytes in cattle may vary depending on a \ ariety of factors that
include pathogens, tissues and time after infection. We believe that developing an
understanding of the role of cytokines in disease can be a step towards controlling infectious
disease.