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. 1969 Oct;17(4):603–615.

The secondary anti-erythrocte response of rabbit spleen cells stimulated in vitro*

Marvis Richardson, J W Moorhead, D L Reedy
PMCID: PMC1455845  PMID: 4187482

Abstract

Suspensions of spleen cells from rabbits immunized to sheep erythrocytes and stimulated in vitro by the specific antigen produced high numbers of antibody-synthesizing cells. Several factors were found to affect secondary antigenic stimulation in vitro. The extent of the immune response elicited was a function of antigen concentration. Maximum was obtained by stimulation with 104 to 105 erythrocytes/107 spleen cells; 107 erythrocytes inhibited the response. The maximum usually occurred on days 4–6; glutamine increased the rate at which antibody-synthesizing cells appeared. It was found that the source of the normal serum supplement used in the growth medium markedly affected the results. Storage of rabbit serum reduced its efficacy; after 6 months storage at –10° little or no response was obtained. The addition of specific antiserum on days 1, 2 or 3 reduced the cellular response.

With cells from twenty-six rabbits tested 6–39 months after immunization, maxima of 1700–29,700 antibody-synthesizing cells/107 spleen cells were attained. No relationship was evident between the time from immunization and the extent of the response. The activity of the extracellular antibody produced by the cells correlated roughly with the number of antibody-synthesizing cells. The antibody elicited in vitro was specific and stable on storage at –10° for 2 years. On the basis of sensitivity to 2-mercaptoethanol, it was a macroglobulin. Disc electrophoresis and identification of the haemolytic component showed that the antibody synthesized in vitro migrated with the γ-globulins, at the same rate as the active component in anti-erythrocyte rabbit serum.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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