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  • Title: Serum factors affecting the specificity of anticardiolipin antibodies.
    Author: Layton GT, Johnston SC, Bertwistle NG.
    Journal: Immunol Cell Biol; 1991 Feb; 69 ( Pt 1)():17-25. PubMed ID: 1869286.
    Abstract:
    The effects were investigated of two pretreatments of human serum and plasma test samples on their subsequent reactivity in the anticardiolipin antibody enzyme-linked immunosorbent assay (ACA-ELISA). The first treatment involved heat inactivation of test samples at 56 degrees C for 30 min, a process sometimes used to inactivate samples from suspected human immunodeficiency virus positive individuals. Such treatment significantly increased the IgG ACA unit/mL values of normal sera, but when this effect was examined further, it was found that the increase in binding occurred on both cardiolipin-coated and uncoated wells and was therefore non-specific. Heat inactivation of sera prior to ACA testing should therefore be avoided. The second treatment involved diluting immunoglobulin (Ig)G and IgM ACA-positive sera in normal human serum (NHS) or newborn calf serum (NCS); sera diluted in NHS showed a significant increase in titre, particularly IgM ACA-positive sera. This phenomenon was found to be due to a serum cardiolipin-binding cofactor which enhances antibody recognition. The cofactor is heat stable and is present in normal sera (male and female) and also in IgG ACA-positive sera. The binding of a human IgM monoclonal antibody to cardiolipin was not affected by the cofactor. The cardiolipin/cofactor complex may represent the optimal autoantigen/autoimmunogen and a re-appraisal, therefore, of the clinical relevance of antibodies to cardiolipin and other negatively charged molecules is warranted.
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