A flow cytometric assay for measuring complement receptor 1 (CR1) and the complement fragments C3d and C4d on erythrocytes

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Abstract

A flow cytometric assay (FCA) was developed to measure complement receptor 1 (CR1) and the complement fragments C3d and C4d on erythrocytes. It was possible to measure these parameters accurately with intra- and interassay coefficients of variation of 2.0% and 6.5% respectively. The method was able to discriminate between low and high levels of erythrocyte CR1, C3d and C4d. Comparison with a previously described RIA method gave excellent correlation coefficients with r2 values of 0.94, 0.93 and 0.91 for CR1, C3d and C4d respectively. The flow cytometric assay was used to measure CR1, C3d and C4d on the erythrocytes of 98 healthy individuals and the 95% upper limits for C3d and C4d were established. There was a wide distribution of CR1 levels amongst these individuals but their C3d and C4d levels were low and often not above background. The possible application of this method in clinical medicine is discussed.

References (27)

  • H.H. Jepsen et al.

    Zinc ions inhibit factor I-mediated release of CR1-bound immune complexes and degradation of cell-bound complement factors C3b and C4b

    Scand. J. Immunol.

    (1990)
  • R.P. Kimberly et al.

    In vivo handling of soluble complement fixing ab/dsDNA immune complexes in chimpanzees

    J. Clin. Invest.

    (1989)
  • N. Madi et al.

    Immune adherence of nascent hepatitis B surface antigen-antibody complexes in vivo in humans

    Clin. Exp. Immunol.

    (1989)
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