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An Effect of Maintenance in Tissue Culture on Spleen Cells Transferred from Antigen-Injected Donors to Syngeneic Mice¹

From The Children's Hospital of Philadelphia and the Department of Pediatrics, University of Pennsylvania, School of Medicine

Abstract

Spleen cells from primed mice incubated with antigen in vitro for transfer to syngeneic mice showed a 4-day interval between cell-transfer and the appearance of antibody at a given threshold level in the recipient's serum, whether the cells were transferred directly or maintained in culture for periods of 1 to 4 days prior to transfer. When spleen cells from antigen-injected donors were transferred directly, the lag period in the recipients varied, being less than 4 days by the number of days between injection of antigen into the donor and collection of its spleen cells (the "donor interval"). However, when such cells were cultured in vitro before transfer, the lag period was constant relative to the time of transfer, 4 days, regardless of the donor interval. It appeared that the progress made in the donors' tissues toward the synthesis of antibody was nullified in tissue culture, so that no matter how many days of the total expected lag period had passed in the donor, the process began anew after in vitro culture and required 4 more days in the recipients' tissues.

The minimum time in culture required for the production of this effect was between 12 and 24 hr. The occurrence of the effect was not constant; the serum or serum fraction used in the medium was a factor.

Where this effect occurred, the cultured cells produced less antibody than control suspensions when the donor interval was 2 days, or longer, whereas no such difference was found with a 1-day donor interval or when the cells had been incubated with antigen in vitro. It is suggested that under the present conditions of tissue culture the processes of maturation or cell division by which antibody-producing cells are produced from their precursors could not occur in culture, as they can in the tissues of the donor or the recipient, and that cells already involved in antibody formation died in tissue culture, so that the processes of maturation or cell division necessarily began again in the recipient animal.

Footnotes

This study was supported by Research Grants HE 04598 and 5 T1 AI 154 of the National Heart Institute and National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Public Health Service.