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[Study on CH50 levels in factor D-depleted serum].

Emi Uchibori1, Etsuko Kitano, Takao Tsuji

  • 1Department of Clinical Laboratory Science, Osaka Prefecture College of Health Sciences, Habikino 583-8555.

Rinsho Byori. the Japanese Journal of Clinical Pathology
|October 11, 2002
PubMed
Summary
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Serum whole complement activity (CH50) assays may not be influenced by alternative pathway (AP) activation. This study demonstrates that CH50 levels remain unaffected even when AP activation is inhibited during the assay.

Area of Science:

  • Immunology
  • Biochemistry

Background:

  • Serum whole complement activity (CH50) assays typically measure classical pathway (CP) function using sensitized sheep erythrocytes.
  • Simultaneous activation of the alternative pathway (AP) during CH50 assays has been proposed, potentially confounding results.

Purpose of the Study:

  • To investigate the influence of AP activation on CH50 levels during the assay.
  • To determine if CH50 measurements are solely reflective of CP activity or if AP contributes.

Main Methods:

  • Developed factor D-depleted serum using polystyrene beads with polyanion (PAMPS-beads).
  • Assayed CH50 levels in normal human serum (NHS) before and after PAMPS-beads treatment.
  • Utilized immunoblot analysis to confirm factor D depletion.

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Main Results:

  • PAMPS-beads treatment successfully depleted factor D, rendering the serum incapable of AP activation (ACH50 undetectable).
  • CH50 levels remained unchanged in serum treated with PAMPS-beads, indicating no significant AP contribution.
  • Re-addition of factor D restored ACH50 levels, validating the depletion method.

Conclusions:

  • The CH50 assay is not significantly affected by alternative pathway activation.
  • CH50 levels accurately reflect classical pathway activity, independent of simultaneous AP activation.