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Studies on the Transfer of Lymph Node Cells

VI. Transfer of Cells Incubated in Vitro with Shigella-Treated Rabbit Serum¹

From The Children's Hospital of Philadelphia (Department of Pediatrics, University of Pennsylvania) and the Division of Immunology, Department of Public Health and Preventive Medicine, Graduate School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania

Abstract

Lymph node cells obtained from donor rabbits not injected with homologous antigen were incubated in vitro with rabbit serum in which Shigella paradysenteriae had been incubated. The cells were washed and then injected into X-irradiated recipient rabbits. On the fourth day, and occasionally on the third day, after transfer, agglutinins to dysentery bacilli appeared in the sera of the recipients. This did not occur when the lymph node cells were heated prior to being transferred. Agglutinins also appeared when untreated lymph node cells and treated serum were injected into the recipients separately.

Within this system the following observations were made:

1. The treated serum was an effective source of antigen in the range of undiluted to 100-fold dilution.

2. The effect described occurred within the range of 14 million to 225 million lymph node cells, with a general correlation between the numbers of cells transferred and the mean agglutinin titer found subsequently.

3. Incubation of cells and treated serum for 30 minutes at 37°C was found to be optimal. Agglutinins did not appear in the sera of recipients when cells and treated serum had been incubated at icebath temperature for 10 minutes, and appeared in low titer after 30 minutes' incubation in an icebath.

4. Agglutinins appeared after the transfer of cells of the mesenteric lymph nodes which had been incubated with treated serum, but not after the transfer of cells of the thymus.

Agglutinins to dysentery bacilli appeared in the sera of non-irradiated recipients after the transfer of untreated lymph node cells incubated in vitro with treated serum. Occasionally agglutinins also appeared following the transfer of heated cells to such recipients, always in lower titer. It was found that there were no differences in titer when the lymph node cells were heated either prior to or after incubation with the treated serum.

Footnotes

¹ This study was supported by a research grant from the National Microbiological Institute of the National Institutes of Health, Public Health Service (G-4104).