Generation of Anti-Boa Immunoglobulin Antibodies for Serodiagnostic Applications, and Their Use to Detect Anti-Reptarenavirus Antibodies in Boa Constrictor

Immunoglobulins (Igs), the key effectors of the adaptive immune system, mediate the specific recognition of foreign structures, i.e. antigens. In mammals, IgM production commonly precedes the production of IgG in the response to an infection. The reptilian counterpart of IgG is IgY, but the exact ki...

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Bibliographic Details
Published in:	PloS one Vol. 11; no. 6; p. e0158417
Main Authors:	Korzyukov, Yegor, Hetzel, Udo, Kipar, Anja, Vapalahti, Olli, Hepojoki, Jussi
Format:	Journal Article
Language:	English
Published:	United States Public Library of Science 29.06.2016 Public Library of Science (PLoS)
Subjects:	Adaptive systems Animals Anti-antibodies Antibodies Antibodies, Viral - blood Antigens Arenavirus Biology and Life Sciences Birds Boa constrictor Boas (Snakes) Boidae - immunology Centrifugal filtration Cercopithecus aethiops Chelonia mydas Chromatography Constrictions Constrictors Diseases Ethylene glycol Female Horseradish peroxidase Immune response Immune serums Immune system Immunization Immunoblotting Immunoglobulin G Immunoglobulin M Immunoglobulin M - immunology Immunoglobulins Immunoglobulins - immunology Immunosuppression Infections Kinetics Liquid chromatography Male Mammals Medicine and Health Sciences Peroxidase Physiological aspects Polyethylene glycol Properties Proteins Purification Python Rabbits Reagents Research and Analysis Methods RNA polymerase Serologic Tests Snakes Vero Cells Veterinary medicine Virology Viruses United States Switzerland Finland
ISSN:	1932-6203, 1932-6203
Online Access:	Get full text
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Summary:	Immunoglobulins (Igs), the key effectors of the adaptive immune system, mediate the specific recognition of foreign structures, i.e. antigens. In mammals, IgM production commonly precedes the production of IgG in the response to an infection. The reptilian counterpart of IgG is IgY, but the exact kinetics of the reptilian immune response are less well known. Boid inclusion body disease (BIBD), an often fatal disease of captive boas and pythons has been linked to reptarenavirus infection, and BIBD is believed to be immunosuppressive. However, so far, the study of the serological response towards reptarenaviruses in BIBD has been hampered by the lack of reagents. Thus we set up a purification protocol for boa constrictor IgY and IgM, which should also be applicable for other snake species. We used centrifugal filter units, poly ethylene glycol precipitation and gel permeation chromatography to purify and separate the IgM and IgY fractions from boa constrictor serum, which we further used to immunise rabbits. We affinity purified IgM and IgY specific reagents from the produced antiserum, and labelled the reagents with horseradish peroxidase. Finally, using the sera of snakes with known exposure to reptarenaviruses we demonstrated that the newly generated reagents can be utilised for serodiagnostic purposes, such as immunoblotting and immunofluorescent staining. To our knowledge, this is the first report to show reptarenavirus-specific antibodies in boa constrictors.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Competing Interests: The authors have declared that no competing interests exist. Conceived and designed the experiments: JH YK OV UH AK. Performed the experiments: JH YK. Analyzed the data: JH YK OV UH AK. Contributed reagents/materials/analysis tools: JH YK OV UH AK. Wrote the paper: JH YK OV UH AK.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0158417