Generation and Characterization of Antibodies against Asian Elephant (Elephas maximus) IgG, IgM, and IgA

Humphreys, Alan F.; Tan, Jie; Peng, RongSheng; Benton, Susan M.; Qin, Xiang; Worley, Kim C.; Mikulski, Rose; Chow, Dar‐Chone; Palzkill, Timothy; Ling, Paul D.

doi:10.1371/journal.pone.0116318

Cited by 16 publications

(13 citation statements)

References 19 publications

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“…Although concentrations of SAA increased by seven days post-vaccination, the cytokine response was not detected until at least 12 days following vaccination, with anti-inflammatory IL-10 being the last to increase. This is a similar timeline to that reported by Humphreys et al [ 78 ], where serum anti-IgG titers to tetanus toxoid increased two to three weeks following vaccination.…”

Section: Discussionsupporting

confidence: 90%

Serum Health Biomarkers in African and Asian Elephants: Value Ranges and Clinical Values Indicative of the Immune Response

Edwards

Miller

Siegal-Willott

et al. 2020

Animals

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Serum biomarkers indicative of inflammation and disease can provide useful information regarding host immune processes, responses to treatment and prognosis. The aims of this study were to assess the use of commercially available anti-equine reagents for the quantification of cytokines (tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukins (IL) 2, 6, and 10) in African (Loxodonta africana, n = 125) and Asian (Elephas maximus, n = 104) elephants, and alongside previously validated anti-human reagents for acute-phase proteins (serum amyloid A and haptoglobin), calculate species-specific biomarker value ranges. In addition, we used opportunistically collected samples to investigate the concentrations of each biomarker during identified clinical cases of illness or injury, as a first step to understanding what biomarkers may be useful to managing elephant health. Immune biomarkers were each elevated above the calculated species-specific value ranges in at least one clinical case, but due to variability in both clinical and non-clinical samples, only serum amyloid A was significantly higher in clinical compared to non-clinical paired samples, with tendencies for higher TNF-α and IL-10. We also detected increased secretion of serum amyloid A and all five cytokines following routine vaccination of a single Asian elephant, indicating that these biomarkers can be beneficial for studying normal immune processes as well as pathology. This study indicates that assays developed with commercial reagents can be used to quantify health biomarkers in wildlife species and identifies several that warrant further investigation to elucidate immune responses to various pathologies.

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Section: Discussionsupporting

confidence: 90%

Serum Health Biomarkers in African and Asian Elephants: Value Ranges and Clinical Values Indicative of the Immune Response

Edwards

Miller

Siegal-Willott

et al. 2020

Animals

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“…Under the non-reducing condition, a single protein band with MW of 150 kDa was visualized. This MW corresponded to the IgG molecule of elephants as previously reported by other groups of investigators [ 26 – 28 ]. The results obtained from 2 elephant serum samples were similar ( Fig 2A ).…”

Section: Resultssupporting

confidence: 84%

Serosurveillance for pandemic influenza A (H1N1) 2009 virus infection in domestic elephants, Thailand

et al. 2017

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The present study conducted serosurveillance for the presence of antibody to pandemic influenza A (H1N1) 2009 virus (H1N1pdm virus) in archival serum samples collected between 2009 and 2013 from 317 domestic elephants living in 19 provinces situated in various parts of Thailand.To obtain the most accurate data, hemagglutination-inhibition (HI) assay was employed as the screening test; and sera with HI antibody titers ≥20 were further confirmed by other methods, including cytopathic effect/hemagglutination based-microneutralization (microNT) and Western blot (WB) assays using H1N1pdm matrix 1 (M1) or hemagglutinin (HA) recombinant protein as the test antigen. Conclusively, the appropriate assays using HI in conjunction with WB assays for HA antibody revealed an overall seropositive rate of 8.5% (27 of 317). The prevalence of antibody to H1N1pdm virus was 2% (4/172) in 2009, 32% (17/53) in 2010, 9% (2/22) in 2011, 12% (1/8) in 2012, and 5% (3/62) in 2013. Notably, these positive serum samples were collected from elephants living in 7 tourist provinces of Thailand. The highest seropositive rate was obtained from elephants in Phuket, a popular tourist beach city. Young elephants had higher seropositive rate than older elephants.The source of H1N1pdm viral infection in these elephants was not explored, but most likely came from close contact with the infected mahouts or from the infected tourists who engaged in activities such as elephant riding and feeding. Nevertheless, it could not be excluded that elephant-to-elephant transmission did occur.

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“…Immunoblot assays. Serum samples were mixed with 2ϫ sample loading buffer, and proteins were resolved by SDS-PAGE as described previously (30). Some of the gels were analyzed by staining with PageBlue protein-staining solution according to the manufacturer's recommendations (ThermoFisher).…”

Section: Methodsmentioning

confidence: 99%

“…Some of the gels were analyzed by staining with PageBlue protein-staining solution according to the manufacturer's recommendations (ThermoFisher). Proteins from a duplicate gel were transferred to nitrocellulose, and immunoblotting was performed with an anti-elephant IgG antibody as described previously (30). For analyses of EEHV Gaussia fusion proteins, supernatants or extracts from cell-associated Gluc fusions were mixed with 2ϫ sample loading buffer and resolved by SDS-PAGE, and immunoblotting was carried out as described previously (30).…”

Section: Methodsmentioning

confidence: 99%

“…Proteins from a duplicate gel were transferred to nitrocellulose, and immunoblotting was performed with an anti-elephant IgG antibody as described previously (30). For analyses of EEHV Gaussia fusion proteins, supernatants or extracts from cell-associated Gluc fusions were mixed with 2ϫ sample loading buffer and resolved by SDS-PAGE, and immunoblotting was carried out as described previously (30). A commercially available rabbit polyclonal anti-Gaussia antibody (New England Biolabs) was used to detect the EEHV-Gluc fusion proteins.…”

Section: Methodsmentioning

confidence: 99%

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Lethal Hemorrhagic Disease and Clinical Illness Associated with Elephant Endotheliotropic Herpesvirus 1 Are Caused by Primary Infection: Implications for the Detection of Diagnostic Proteins

Fuery

Pursell

Tan

et al. 2020

J Virol

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Elephant endotheliotropic herpesvirus (EEHV) can cause lethal hemorrhagic disease in juvenile Asian elephants, both in captivity and in the wild. Most deaths associated with the virus are caused by two chimeric variants of EEHV1 (EEHV1A and EEHV1B), while two other EEHVs endemic within Asian elephants (EEHV4 and EEHV5) have been recognized but cause death less often. Whether lethal EEHV infections are due to primary infection or reactivation of latent virus remains unknown, and knowledge of the anti-EEHV antibody levels in young elephants is limited. To close these gaps, we sought to develop a serologic assay capable of distinguishing among infections with different EEHVs using a luciferase immunoprecipitation system (LIPS) for antibody profiling and a panel of conserved EEHV recombinant proteins and proteins unique to EEHV1. The results showed that elephants dying from EEHV1 hemorrhagic disease or ill from EEHV infection were seronegative for the EEHV species that caused the disease or illness, indicating that the events were associated with primary infection rather than reactivation of latent virus. We also demonstrated that waning of EEHV1-specific antibodies can occur in the first 2 years of life, when a threshold protective level of antibody may be needed to prevent severe EEHV1-related disease. Use of the LIPS assay to identify putative “diagnostic” proteins would be a valuable asset in determining the EEHV immune status of young elephants and responses to candidate EEHV vaccines in the future. IMPORTANCE Whether clinical illness and deaths associated with elephant endotheliotropic herpesvirus (EEHV) infection result from primary infection or reactivation of latent virus is a longstanding question in the field. By applying a relatively new assay, the luciferase immunoprecipitation system (LIPS), combined with the genomic sequences of the viruses, we gained the insights and tools needed to resolve this issue. Our EEHV1-specific LIPS assay should be useful for assessing the vulnerability of elephant calves to infection with different EEHVs and evaluating antibody responses to anti-EEHV vaccines. A significant proportion of the Asian elephant population is under some form of human care. Hence, the ability to screen for EEHV immune status in elephant calves should have a major impact on the management of these animals worldwide.

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Generation and Characterization of Antibodies against Asian Elephant (Elephas maximus) IgG, IgM, and IgA

Cited by 16 publications

References 19 publications

Serum Health Biomarkers in African and Asian Elephants: Value Ranges and Clinical Values Indicative of the Immune Response

Serum Health Biomarkers in African and Asian Elephants: Value Ranges and Clinical Values Indicative of the Immune Response

Serosurveillance for pandemic influenza A (H1N1) 2009 virus infection in domestic elephants, Thailand

Lethal Hemorrhagic Disease and Clinical Illness Associated with Elephant Endotheliotropic Herpesvirus 1 Are Caused by Primary Infection: Implications for the Detection of Diagnostic Proteins

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