An indirect flow cytometric test for detection of anti-neutrophil antibodies in dogs.

Am J Vet Res

Department of Veterinary and Biomedical Sciences, College of Veterinary Medicine, University of Minnesota, Saint Paul, MN 55108, USA.

Published: May 2007

AI Article Synopsis

  • Developed an assay to detect anti-neutrophil antibodies in dogs using serum samples from healthy and sick dogs.
  • Involved indirect immunofluorescence testing to assess neutrophil fluorescence intensity and percentage of neutrophils, showing no significant differences between healthy and sick dogs, except in one case with steroid-responsive neutropenia.
  • Results indicate the assay is consistent and reliable, suggesting potential for future evaluation in neutropenic dogs and response to immunosuppressive treatment.

Article Abstract

Objective: To develop a clinically applicable assay for detection of serum anti-neutrophil antibodies in dogs.

Sample Population: Serum samples of 20 healthy dogs and 20 sick dogs.

Procedures: An indirect immunofluorescence assay was developed in which canine serum was incubated with paraformaldehyde-fixed neutrophils and subsequently incubated with fluorescein-conjugated rabbit anti-dog IgG. Neutrophil median fluorescence intensity and the percentage of neutrophils with an increase in fluorescence intensity were determined by use of a flow cytometer.

Results: Neutrophils incubated with serum from healthy and sick dogs had a normally distributed curve when displayed as a histogram. Alloantibodies or immune complexes that significantly affected test results were not detected. Hyperglobulinemia did not appear to affect test results. The neutrophil donor did not significantly affect test results. With 1 exception, results for the sick dogs did not differ appreciably from those for healthy dogs. Serum from a dog with steroid-responsive neutropenia had a greater neutrophil fluorescence value and percentage of neutrophils with an increase in fluorescence intensity, compared with either healthy or sick dogs.

Conclusions And Clinical Relevance: The indirect immunofluorescence test gave consistent results for healthy and sick dogs and detected anti-neutrophil antibodies in a dog with steroid-responsive neutropenia. Definitive evaluation of the test will be dependent on evaluation of persistently neutropenic dogs and correlation of test results with a response to immunosuppressive therapy.

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Source
http://dx.doi.org/10.2460/ajvr.68.5.464DOI Listing

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