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Kinetic Studies on the Incidence of Cells Appearing to form Two Antibodies¹

From the Department of Genetics, Stanford University School of Medicine, Palo Alto, California

Abstract

The question of whether any cell can simultaneously form two or more antibodies has been reinvestigated with better and more complete techniques of antibody assay than previously described. Rats were given injections of flagellin from one or two strains of Salmonella containing significant amounts of both H and O antigens. The dose used at each injection varied from 0.002 µg to 6 mg. Injections were given once a week with 4-week rests after 4 injections, or once a month. The duration of immunization was 1 month to 18 months. Most rats started their immunization course in adult life, but some on the day of birth. Antibody released into the microdrop surrounding each single cell was detected by H immobilization, O agglutination, and the chaining phenomenon. Antibody attached to the surface of the cell was detected by H or O adherence. Of 3628 single cells tested, 41 appeared to be doubly active; but only two of these were shown to actually release two antibodies simultaneously, the majority of the others releasing one antibody and having a second reactivity present on the surface. Thus the incidence of "true double producers" was less than 0.01%. No experimental variation of antigen dose or immunization schedule seemed to affect this proportion.

The validity of the experimental methods used and the possible theoretical basis for the observed results are discussed.

Footnotes

This work was supported by Grant E-2700 from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Public Health Service, Bethesda, Maryland.

² Present address: Walter and Eliza Hall Institute, Melbourne, Australia.

³ Present address: Department of Serology and Bacteriology, University of Helsinki, Helsinki, Finland.

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