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In vitro Antibody Synthesis by Diffusion Chamber Cultures of Spleen Cells

II. Effect of Increased Levels of Free Antibody¹

From the Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee

Abstract

The effect of free antibody on antibody production in the secondary immune response has been studied by use of antigen-stimulated primed spleen cells in diffusion chamber cultures. The cultures were exposed to high concentrations of antibody at various times after the cultures were begun, and antibody production was assayed by determining the amounts of in vitro radioactive amino acid incorporation into specific antibody. Chamber cultures implanted from the start in animals hyperimmunized to the test antigen (sheep red blood cells) failed to exhibit any significant antibody production. When the cultures were grown for 7 to 9 days in normal animals and exposed to free antibody either at the time of in vitro incubation or by transfer into hyperimmune animals for 1 or 2 days prior to the in vitro assessment of antibody production, there was no significant depression of antibody production. This was true even if the cultures were exposed to kiloroentgen doses of x-ray at the beginning of the plateau phase of their immune response to eliminate turnover in the antibody-producing cell population. These results suggest that antibody-mediated regulation of antibody production occurs at the level of the antibody-producing cell population, possibly by preventing antigenic determinants from triggering progenitor cells that participate in the immune response, and not through control of the synthetic or secretory activities of antibody-producing cells.

Footnotes

¹ Research sponsored by the U. S. Atomic Energy Commission under contract with the Union Carbide Corporation.

² Present address: Department of Experimental Pathology, Scripps Clinic and Research Foundation. La Jolla, California 92037.