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Chemiluminescence and Chemotaxis Assay of Canine Granulocytes: A Methodological Study

Chemiluminescence och kemotaxis: en metodologisk studie avseende hundens granulocyter.

Acta Veterinaria Scandinavica volume 31, pages 79–86 (1990)Cite this article

Abstract

Chemiluminescence (CL) of isolated granulocytes and of whole blood from dogs was evaluated. Chemiluminescence of whole blood samples created an undesired quenching effect by the red blood cells which makes the assay difficult to apply in pathological cases with low formation of oxygen metabolites. This problem was avoided when chemiluminescence was determined, using isolated granulocytes. A cell concentration of 5×109/1 was needed to create optimal conditions. The Boyden chamber technique was used for study of random migration and Chemotaxis. Casein (0.1 %), zymosan activated serum with and without e-amino-n-caproic-acid and homologous serum were effective chemoattractants for canine granulocytes, while FMLP (formyl-methionyl-leucyl-phenylalanin) did not attract canine granulocytes.

Sammanfattning

Detta arbete omfattar en evaluering av en metod at mäta granulocytes förmåga till avdödning, s k chemiluminescence (CL) samt en utvärdering av olika kemotaktiska substanser i en Boyden kammarmetod för mätning av granulocytes kemotaxis hos hund.

Med avsikt att utveckla en snabbare netod för mätning av CL, användes helblod som jämfördes med isolerade celler från samma individ. Responsen blev dock så dämpad att det ej skulle vara möjligt bedöma en eventuell funktionsstörning med för låg chemiluminescence i den kliniska situationen. Olika cellantal testades och 5 × 109 celler/l bedömdes som en lämplig koncentration då den gav tillräckligt stort utslag för att en underfunktion skulle kunna diagnostiseras. Temperaturens inverkan på responsen undersöktes genom att mäta CL vid 37° och 39°C men det förelag ej någon signifikant skillnad mellan mätningarna. Lagring av celler längre än 3 timmar ger signifikant lägre chemiluminescence. Det är således viktigt att man standardiserar tiden mellan cellisolering och mätning av CL.

Olika kemotaktiska substanser (casein, zymosan-aktiverat serum med och utan ε-kapronsyra, homologt serum och formyl-methionyl-leucocyl-phenylalanin, (FMLP)) användes för att stimulera granulocyternas kemotaxis och jämfördes med ej stimulerad migration där cellerna fick vandra mot buffert. Samtliga kemotaktiska agens utom FMLP attraherade hundgranulocyter i detta system och kan följaktligen användas i en klinisk situation för att diagnosticera patologiska tillstånd.

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Authors and Affiliations

  1. Department of Medicine and Surgery, Veterinary Faculty, Swedish University of Agricultural Sciences, Uppsala, Sweden

    Gunilla Trowald-Wigh & Kerstin Thorén- Tolling

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  1. Gunilla Trowald-Wigh
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  2. Kerstin Thorén- Tolling
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Trowald-Wigh, G., Thorén- Tolling, K. Chemiluminescence and Chemotaxis Assay of Canine Granulocytes: A Methodological Study. Acta Vet Scand 31, 79–86 (1990). https://doi.org/10.1186/BF03547580

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Acta Veterinaria Scandinavica

ISSN: 1751-0147