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Benzylpenicilloyl-Specific Serum Antibodies to Penicillin in Man

II. Sensitivity of the Hemagglutination Assay Method, Molecular Classes of the Antibodies Detected, and Antibody Titers of Randomly Selected Patients¹

From the Department of Medicine, New York University School of Medicine, New York, New York

Abstract

The molecular classes of benzylpenicilloylspecific (BPO) antibodies detected in human sera by the presently described passive hemagglutination method (HA) were studied by mercaptoethanol (ME) degradation, ultracentrifugal fractionation in sucrose, and neutralization with monospecific rabbit anti-immunoglobulin sera. Sera whose HA titers were reduced to zero by ME treatment were shown to contain virtually only IgM antibodies. Sera whose HA titers were not reduced by ME were shown to contain mainly IgG antibodies, but small amounts of IgM and antibodies of other molecular classes were also present.

The HA method was capable of detecting as little as 0.0003 µg/ml anti-benzylpenicilloyl (BPO) antibody protein in rabbit antisera and it may be more sensitive for human sera. By contrast, the passive cutaneous anaphylaxis (PCA) test in guinea pigs was 200-fold less sensitive for rabbit sera and 1500-fold for human sera.

By means of this assay method, BPO-specific antibodies were detected in the sera of 97% of patients selected at random; most sera contained only IgM antibodies and 14% contained IgG antibodies in addition to IgM. BPO specificity of these antibodies was proved by hapten inhibition.

Because of its specificity, sensitivity and simplicity, this method may be applicable to diverse immunologic studies.

Footnotes

¹ Supported by the Health Research Council of the City of New York under Contract U-1297, and the United States Public Health Service under Grant No. A-2594.

² Career Investigator of the Health Research Council of the City of New York.

³ Dermatology Training Fellow under Grant No. T1 AM 5326 from the National Institute of Arthritis and Metabolic Diseases, United States Public Health Service.

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