Studies on the immune response to Newcastle disease virus in poultry

162 Pages·2017·3.56 MB·English

Iowa State University Capstones, Theses and
Retrospective Theses and Dissertations
Dissertations
1996
Studies on the immune response to Newcastle
disease virus in poultry
Anwar Diab Maraqa
Iowa State University
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Recommended Citation
Maraqa, Anwar Diab, "Studies on the immune response to Newcastle disease virus in poultry " (1996).Retrospective Theses and
Dissertations. 11165.
https://lib.dr.iastate.edu/rtd/11165
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Studies on the immune response to Newcastle
disease virus in poultry
by
Anwar Diab Maraqa
A Dissertation Submitted to the
Grraduate Faculty in Partial Fulfillment of the
Requirements for the Degree of
DOCTOR OF PHILOSOPHY
Department: Microbiology, Immunology and Preventive Medicine
Major: Veterinary Microbiology
Approved:
Signature was redacted for privacy.
In Charge of Miajor Work
Signature was redacted for privacy.
Signature was redacted for privacy.
For the Major Department
Signature was redacted for privacy.
F
Iowa State University
Ames, Iowa
1996
UMI Number: 9626053
IMI Microform 9626053
Copyright 1996, by UMI Company. All rights reserved.
This microform edition is protected against unauthorized
copying under Title 17, United States Code.
UMI
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a
This dissertation is dedicated to my wife,
Kholoud Maraqa
Thank you for your love, prayers and encouragement.
Ill
TABLE OF CONTENTS
GENERAL INTRODUCTION 1
Dissertation Research Objectives 2
Dissertation Organization 2
LITERATURE REVIEW 4
Newcastle Disease 4
History 4
Etiologic agent 5
Classification 5
Properties of the virion 5
Properties of the genome 6
Properties of the virus proteins 6
Replication 9
Cultivation 9
The disease 10
Hosts 10
Transmission 11
Pathogenesis 11
Pathotype and pathogenicity 12
Diagnosis 13
Serology 13
Prevention and control 14
Vaccination 14
Live NDV vaccines 14
Inactivated vaccines 15
Recombinant NDV vaccines 16
Immunity to Newcastle Disease 16
Humoral immunity 16
Maternally derived antibodies 17
Local immunity 18
The role of NDV polypeptides in protection 19
Ceil mediated immunity 21
Cell mediated immunity in other viruses 23
Cyclophosphamide 24
A TECHNIQUE FOR INDUCING B-CELL ABLATION IN CHICKENS BY
IN OVO INJECTION OF CYCLOPHOSPHAMIDE 27
iv
SUMMARY 27
INTRODUCTION 29
MATERIALS AND METHODS 30
Eggs 30
Media and reagents 31
Cyclophosphamide treatment 32
Virus 32
Virus propagation and purification 32
Whole blood 33
Colorimetric blastogenesis assay 33
Flow cytometry analysis 34
Hemagglutination inhibition (HI) test 3 5
Experimental design 36
Statistical evaluation 37
RESULTS 37
Effect of in ovo cyclophosphamide treatment on hatchability, mortality,
and body weight 37
Whole blood blastogenesis microassay 38
Antibody response to Newcastle disease vaccine 39
Effect of CY on T and B-cell populations in PBL 41
DISCUSSION 43
REFERENCES 46
ACKNOWLEDGMENTS 48
APPENDIX: DATA AND ANALYSIS 49
STUDIES ON NEWCASTLE DISEASE IN CHICKENS: THE ROLE OF CELL
MEDIATED IMMUNITY IN PROTECTION 52
SUMMARY 52
INTRODUCTION 55
MATERIALS AND METHODS 56
Eggs 56
Media and reagents 57
V
Virus propagation and purification 58
Preparation of purified lymphocytes 59
Cyclophosphamide treatment 59
Colorimetric blastogenesis assay 60
Western blot 61
Sample collection for virus isolation 62
Hemagglutination inhibition (HI) test 62
Virus neutralization (VN) test 63
Experimental design 63
Statistical evaluation 65
RESULTS 65
Humoral immune response 65
Blastogenesis response 67
Challenge results 69
DISCUSSION 74
REFERENCES 78
ACKNOWLEDGMENTS 80
APPENDIX; DATA AND ANALYSIS 81
THE ROLE OF ANTIBODIES TO NEWCASTLE DISEASE VIRUS PROTEINS
IN PROTECTION 90
SUMMARY 90
INTRODUCTION 92
MATERIALS AND METHODS 94
Embryonated eggs and chickens 94
Media and reagents 94
Virus strains 94
Virus propagation and purification 94
Preparation of NDV proteins 95
Preparation of antiserum 96
ELISA 96
Western blot 97
Hemagglutination inhibition (HI) test 98
Virus neutralization (VN) test 98
vi
Sample collection for virus isolation 99
Experimental design 99
RESULTS 100
DISCUSSION 105
REFERENCES 108
ACKNOWLEDGMENTS 110
A RAPID VIRUS ^^^EUTRALIZATION ASSAY FOR NEWCASTLE DISEASE
VIRUS USING THE SWINE TESTICULAR (ST) CONTINUOUS CELL LINE 111
SUMMARY 111
INTRODUCTION 113
MATERIAL AND METHODS 114
Media and regents 114
Viruses 115
Cell culture 115
Virus inoculation 116
Serum samples 116
Virus titration in CEF and ST cells 117
Virus neutralization (VN) assay 117
Serum effect on the MTT VN microassay 118
Statistical evaluation 119
RESULTS 120
Growth of Texas GB and B1 strains ofNDV in CEF and continuous
cell lines 120
Titration of Texas GB strain of NDV in CEF and ST 123
Virus neutralization assay 123
Serum effect 123
DISCUSSION 126
REFERENCES 129
ACKNOWLEDGMENTS 130