Generation and characterization of antibodies against Asian elephant (Elephas maximus) IgG, IgM, and IgA.

TitleGeneration and characterization of antibodies against Asian elephant (Elephas maximus) IgG, IgM, and IgA.
Publication TypeJournal Article
Year of Publication2015
AuthorsHumphreys, AF, Tan, J, Peng, R, Benton, SM, Qin, X, Worley, KC, Mikulski, RL, Chow, D-C, Palzkill, TG, Ling, PD
JournalPLoS One
Volume10
Issue2
Paginatione0116318
Date Published2015
ISSN1932-6203
KeywordsAnimals, Antibodies, Anti-Idiotypic, Base Sequence, Betaherpesvirinae, Chromatography, Gel, DNA Primers, Electrophoresis, Polyacrylamide Gel, Elephants, Endangered Species, Enzyme-Linked Immunosorbent Assay, Herpesviridae Infections, Immunoglobulin A, Immunoglobulin G, Immunoglobulin M, Molecular Sequence Data, Polymerase Chain Reaction, Sequence Analysis, DNA, Tandem Mass Spectrometry
Abstract

Asian elephant (Elephas maximus) immunity is poorly characterized and understood. This gap in knowledge is particularly concerning as Asian elephants are an endangered species threatened by a newly discovered herpesvirus known as elephant endotheliotropic herpesvirus (EEHV), which is the leading cause of death for captive Asian elephants born after 1980 in North America. While reliable diagnostic assays have been developed to detect EEHV DNA, serological assays to evaluate elephant anti-EEHV antibody responses are lacking and will be needed for surveillance and epidemiological studies and also for evaluating potential treatments or vaccines against lethal EEHV infection. Previous studies have shown that Asian elephants produce IgG in serum, but they failed to detect IgM and IgA, further hampering development of informative serological assays for this species. To begin to address this issue, we determined the constant region genomic sequence of Asian elephant IgM and obtained some limited protein sequence information for putative serum IgA. The information was used to generate or identify specific commercial antisera reactive against IgM and IgA isotypes. In addition, we generated a monoclonal antibody against Asian elephant IgG. These three reagents were used to demonstrate that all three immunoglobulin isotypes are found in Asian elephant serum and milk and to detect antibody responses following tetanus toxoid booster vaccination or antibodies against a putative EEHV structural protein. The results indicate that these new reagents will be useful for developing sensitive and specific assays to detect and characterize elephant antibody responses for any pathogen or vaccine, including EEHV.

DOI10.1371/journal.pone.0116318
Alternate JournalPLoS One
PubMed ID25658336
PubMed Central IDPMC4320114
Grant ListP30 CA125123 / CA / NCI NIH HHS / United States

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