Identification of specific antinuclear antibodies in dogs using a line immunoassay and enzyme-linked immunosorbent assay

Bremer, Hanna; Lattwein, Erik; Renneker, Stefanie; Lilliehöök, Inger; Rönnelid, Johan; Hansson-Hamlin, Helene

doi:10.1016/j.vetimm.2015.10.002

Cited by 14 publications

(22 citation statements)

References 45 publications

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“…This is significantly lower than the concentration seen in the dogs with lymphoma reported in this study with median of 590 ng/mL. Comparisons between the concentrations of nucleosomes reported here and those in other reports of inflammation and control groups are difficult due to the fact that nucleosome concentrations have previously been reported only in arbitrary units(7)(8)(9)(20)(21)(22).…”

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confidence: 81%

Evaluation of nucleosome concentrations in healthy dogs and dogs with cancer

Wilson-Robles

Miller

Jarvis

et al. 2020

Preprint

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AbstractIntroductionNucleosomes consist of small fragments of DNA wrapped around a histone octamer core. Diseases such as cancer or inflammation lead to cell death, which causes fragmentation and release of nucleosomes into the blood. The Nu.Q™ technology measures circulating nucleosome levels and exploits the different compositions of cancer derived nucleosomes in blood to detect and identify cancer even at early stages. The objectives of this study are to identify the optimal sample type for the Nu.Q™ H3.1 assay and to determine if it can accurately detect nucleosomes in the blood of healthy canines as well as those with cancer.Materials and MethodsBlood samples from healthy canine volunteers as well as dogs newly diagnosed with lymphoma were used. The blood was processed at a variety of times under a variety of conditions to determine the most reliable sample type and conditions, and to develop an appropriate processing strategy to ensure reliably accurate results.ResultsNucleosomes could be detected using a variety of sample collection and processing protocols. Nucleosome signals were highest in EDTA plasma and serum samples and most consistent in plasma. Samples should be processed within an hour of collection. Experiments showed that samples were able to withstand several freeze thaw cycles. Processing time and tcollection tube type did affect nucleosome detection levels. Finally, significantly elevated concentrations of nucleosomes were seen in a small cohort of dogs that had been newly diagnosed with lymphoma.ConclusionsWhen samples are collected and processed appropriately, the Nu.Q™ platform can reliably detect nucleosomes in the plasma of dogs. Further testing is underway to validate and optimize the Nu.Q™ platform for veterinary use.

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contrasting

confidence: 81%

Evaluation of nucleosome concentrations in healthy dogs and dogs with cancer

Wilson-Robles

Miller

Jarvis

et al. 2020

Preprint

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“…Several autoantigens have been previously identified in dogs by different approaches 9 , 21 , 22 , 38 , 46 – 50 . Here, we report the use of human protein arrays for the screening of canine autoantigens.…”

Section: Discussionmentioning

confidence: 99%

“…The standard test for detecting canine ANA – indirect immunofluorescence microscopy (IIF) – uses cells of human origin (HEp-2) as substrate 20 . In a previous study, it could be shown that several autoantibodies with important roles in human disease were also present in dogs 21 . Thus, the autoantibody targets in human and canine disease frequently overlap, although there are some autoantibodies that are species-specific 22 .…”

Section: Introductionmentioning

confidence: 95%

ILF2 and ILF3 are autoantigens in canine systemic autoimmune disease

Bremer

Landegren

Sjöberg

et al. 2018

Sci Rep

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Dogs can spontaneously develop complex systemic autoimmune disorders, with similarities to human autoimmune disease. Autoantibodies directed at self-antigens are a key feature of these autoimmune diseases. Here we report the identification of interleukin enhancer-binding factors 2 and 3 (ILF2 and ILF3) as autoantigens in canine immune-mediated rheumatic disease. The ILF2 autoantibodies were discovered in a small, selected canine cohort through the use of human protein arrays; a method not previously described in dogs. Subsequently, ILF3 autoantibodies were also identified in the same cohort. The results were validated with an independent method in a larger cohort of dogs. ILF2 and ILF3 autoantibodies were found exclusively, and at a high frequency, in dogs that showed a speckled pattern of antinuclear antibodies on immunofluorescence. ILF2 and ILF3 autoantibodies were also found at low frequency in human patients with SLE and Sjögren’s syndrome. These autoantibodies have the potential to be used as diagnostic biomarkers for canine, and possibly also human, autoimmune disease.

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“…Previous work in dogs has reported AHA to be common in dogs suffering from SLE . Besides having a nuclear function, recent studies have indicated a significant toxic or pro‐inflammatory effect of histones when released into extracellular space by damaged and activated cells .…”

Section: Discussionmentioning

confidence: 99%

“…37 Previous work in dogs has reported AHA to be common in dogs suffering from SLE. 38 Besides having a nuclear function, recent studies have indicated a significant toxic or pro-inflammatory effect of histones when released into extracellular space by damaged and activated cells. 39 Histone release has been suggested to play a part in autoimmune disease, 40 not just acting as a direct autoantigen, but also magnifying the destructive autoimmune processes.…”

Section: Discussionmentioning

confidence: 99%

Antihistone Autoantibodies in Dobermans With Hepatitis

Dyggve

Meri

Spillmann

et al. 2017

Veterinary Internal Medicne

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BackgroundImmune system involvement is suggested as an underlying cause for Doberman hepatitis (DH) based on female predisposition, lymphocyte infiltration, abnormal hepatocyte expression of major histocompatibility complex class II antigens, and homozygosity for dog leukocyte antigen DRB1*00601.ObjectiveTo measure serum antinuclear antibodies (ANA) and serum antihistone antibodies (AHA) in Dobermans with hepatitis. To determine whether increased serum ANA or serum AHA could be used to support the diagnosis of Doberman hepatitis (DH).AnimalsPrivately owned 25 subclinically and 13 clinically affected DH Dobermans and 17 healthy control Dobermans.MethodsCase–control study. Indirect immunofluorescence (IIF) microscopy and line blot tests were employed for the ANA pilot studies and an enzyme‐linked immunosorbent assay (ELISA) assay for detection of IgG AHA.ResultsIndirect immunofluorescence revealed ANA‐positive cases, and line blot showed AHA reactivity. In ELISA, importantly increased concentrations of AHA were found in 92% (23/25) of dogs in the subclinical stage and 84.6% (11 of 13) of dogs in the clinical stage of DH compared with no control dogs (0/17) (P < 0.0005). The mean AHA absorbance values of the blood samples obtained from the 25 subclinical DH dogs (1.36 ± 0.60, mean ± SD) and the 13 clinically affected dogs (1.46 ± 0.49) were significantly higher than in 17 control dogs (0.51 ± 0.18; P < 0.0001).Conclusions and Clinical ImportanceAs the presence of AHA indicates autoimmune activity, our results favor an autoimmune background as one cause for DH. Antihistone antibody could represent a novel means for screening Dobermans with increased serum alanine transaminase concentrations and suspicion of DH.

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Identification of specific antinuclear antibodies in dogs using a line immunoassay and enzyme-linked immunosorbent assay

Cited by 14 publications

References 45 publications

Evaluation of nucleosome concentrations in healthy dogs and dogs with cancer

Evaluation of nucleosome concentrations in healthy dogs and dogs with cancer

ILF2 and ILF3 are autoantigens in canine systemic autoimmune disease

Antihistone Autoantibodies in Dobermans With Hepatitis

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