A chemiluminescence assay for erythrophagocytosis

Journal of Bioluminescence and Chemiluminescence
I DowningR H Fraser

Abstract

A luminol-dependent chemiluminescence assay for the assessment of the phagocytosis of erythrocytes sensitized with anti-D IgG immunoglobulin by mononuclear leukocytes is described. The mononuclear leukocytes were obtained by apheresis enriched by centrifugation through a density gradient and stored in liquid nitrogen before use. The total reaction mixture, consisting of mononuclear leukocytes-luminol-erythrocytes (either anti-D IgG sensitized or unsensitized controls) was 500 microliters, light detection was by an LKB 1251 luminometer. Peak luminescence was seen between 35-45 minutes, the reaction being exhausted by 120 minutes. Determination of the reproducibility of the assay gave intra- and inter-assay coefficients of variation of 5% and 13% respectively. We found the chemiluminescent response to be affected by the number of erythrocytes used in the assay and by the composition of the medium in which the cells were resuspended, particularly the pH at the initiation of the assay. We also compared the chemiluminescence assay to a microscopic phagocytic assay and found the results virtually identical. However, the former chemiluminescence assay was much easier to perform, marginally more sensitive, less laborious and eliminated...Continue Reading

References

Jan 1, 1976·Journal of Immunological Methods·R S Weiner
Jan 1, 1978·American Journal of Hematology·S J WeissA F LoBuglio
Jan 1, 1986·Methods in Enzymology·D Roos, M de Boer
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Feb 1, 1966·Biochemistry·N E GoodR M Singh
Jan 1, 1984·Clinical and Laboratory Haematology·A Hadley, A M Holburn

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Citations

Jan 1, 1996·Transfusion clinique et biologique : journal de la Société française de transfusion sanguine·A J StrachanG P Rowe
Nov 25, 2000·British Journal of Haematology·S MiescherF Wurm
Mar 1, 1995·Transfusion Medicine·I M BromilowJ K Duguid
Mar 10, 2007·Journal of Immunological Methods·Geraldine HealeyRosemary L Sparrow
Jan 1, 1991·International Journal of Cardiac Imaging·N G PandianD N Salem

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