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Is serum the optimal source for HLA antibodies?
Vox Sanguinis
|January 1, 1987
Summary
Serum antibodies exhibit superior cytotoxic reactivity compared to anticoagulated preparations. Calcium chelation and pH changes significantly inhibit antibody function, highlighting serum
Area of Science:
- Immunology
- Transfusion Medicine
Background:
- Cytotoxic antibody testing is crucial for HLA antibody detection.
- Anticoagulants are often used in blood sample processing, potentially affecting antibody reactivity.
- Understanding the impact of anticoagulants on antibody-dependent cell-mediated cytotoxicity (ADCC) is vital for accurate diagnostics.
Purpose of the Study:
- To compare the cytotoxic reactivity of human leukocyte antigen (HLA) antibodies in serum versus various anticoagulated plasma preparations.
- To investigate the effects of specific anticoagulants (ACD-A, EDTA, citrate, heparin) on antibody-mediated cytotoxicity.
- To elucidate the mechanisms underlying anticoagulant-induced inhibition, including calcium chelation, pH, and anticomplementary activity.
Main Methods:
- Compared cytotoxic reactivity of 18 class I HLA serum antibodies with antibody preparations in ACD-A, EDTA, citrate, and heparin plasmas.
- Assessed the impact of recalcification and pH adjustment on antibody reactivity in anticoagulated samples.
- Utilized inhibition tests, serial titrations, and varying calcium concentrations to analyze anticoagulant effects.
Main Results:
- All tested anticoagulated plasmas (ACD-A, EDTA, citrate, heparin) demonstrated lower cytotoxic reactivity than serum.
- Calcium chelation by EDTA and citrate significantly inhibited cytotoxicity, an effect more pronounced with pre-incubation.
- pH changes also contributed to reduced reactivity, with adjustments toward serum pH improving function; heparin showed minimal impact but exhibited dose-dependent anticomplementary activity.
Conclusions:
- Serum is the superior source for antibody preparations in cytotoxicity testing due to higher reactivity.
- Calcium chelation and pH alterations are key factors responsible for the reduced antibody function in anticoagulated samples.
- These findings underscore the importance of using serum for consistent and reliable HLA antibody detection in immunological assays.