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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| Published in: | PloS one Vol. 11; no. 6; p. e0158417 |
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| ISSN: | 1932-6203, 1932-6203 |
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| Abstract | 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. |
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| AbstractList | 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. 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.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. |
| Audience | Academic |
| Author | Hetzel, Udo Vapalahti, Olli Kipar, Anja Korzyukov, Yegor Hepojoki, Jussi |
| AuthorAffiliation | Division of Clinical Research, UNITED STATES 1 University of Helsinki, Medicum, Department of Virology, Helsinki, Finland 2 Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland 4 University of Helsinki and Helsinki University Hospital, Department of Virology, Helsinki, Finland 3 University of Helsinki, Department of Veterinary Biosciences, Faculty of Veterinary Medicine, Helsinki, Finland |
| AuthorAffiliation_xml | – name: 4 University of Helsinki and Helsinki University Hospital, Department of Virology, Helsinki, Finland – name: 2 Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland – name: Division of Clinical Research, UNITED STATES – name: 1 University of Helsinki, Medicum, Department of Virology, Helsinki, Finland – name: 3 University of Helsinki, Department of Veterinary Biosciences, Faculty of Veterinary Medicine, Helsinki, Finland |
| Author_xml | – sequence: 1 givenname: Yegor surname: Korzyukov fullname: Korzyukov, Yegor – sequence: 2 givenname: Udo surname: Hetzel fullname: Hetzel, Udo – sequence: 3 givenname: Anja surname: Kipar fullname: Kipar, Anja – sequence: 4 givenname: Olli surname: Vapalahti fullname: Vapalahti, Olli – sequence: 5 givenname: Jussi surname: Hepojoki fullname: Hepojoki, Jussi |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/27355360$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_3390_v12040457 crossref_primary_10_1016_j_cvex_2021_01_007 crossref_primary_10_1128_JVI_01968_20 crossref_primary_10_3390_v14010107 crossref_primary_10_1128_JVI_00718_17 crossref_primary_10_1128_spectrum_01585_22 crossref_primary_10_1242_jeb_214171 crossref_primary_10_1016_j_dci_2024_105302 crossref_primary_10_1371_journal_ppat_1007415 crossref_primary_10_1099_jgv_0_002052 crossref_primary_10_3389_fvets_2021_733404 crossref_primary_10_1099_jgv_0_002100 crossref_primary_10_1371_journal_ppat_1006179 crossref_primary_10_3390_v12040395 crossref_primary_10_1111_avj_12792 crossref_primary_10_1371_journal_pone_0229667 crossref_primary_10_1371_journal_pone_0221863 |
| Cites_doi | 10.1002/elps.1150180309 10.1038/npg.els.0000521 10.1177/026119291604400111 10.3791/3084 |
| ContentType | Journal Article |
| Copyright | COPYRIGHT 2016 Public Library of Science 2016 Korzyukov et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. 2016 Korzyukov et al 2016 Korzyukov et al |
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| Notes | 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. |
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| Title | Generation of Anti-Boa Immunoglobulin Antibodies for Serodiagnostic Applications, and Their Use to Detect Anti-Reptarenavirus Antibodies in Boa Constrictor |
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