Studies on antibody-producing cells. 3. Identification of young plaque-forming cells by thymidine- 3 H labeling.

Mice injected with sheep RBC and then, 4 days later with thymidine-(3)H, were sacrificed on the day of thymidine-(3)H injection or 1 or 2 days later. Hemolytic antibody plaque preparations were made of cells from the draining lymph nodes by a thin-plating procedure permitting collection of isolated PFC for electron microscopic examination and radioautography. Of cells obtained on the day of thymidine-(3)H injections, 65% of the labeled PFC were in the lymphocytic category, in comparison with 13% found previously in the entire population of such cells. The remaining 35% were plasmablasts in early stages of differentiation. Cells obtained 1 day after the thymidine-(3)H injections showed a shift to a majority of labeled cells in the plasmacytic category. Also, the plasmablasts were substantially more differentiated than those of the previous day, and some mature plasma cells were now seen. The labeled PFC obtained on day 2 gave no indication of further differentiation. Cells of rabbit lymph nodes labeled in vitro with thymidine-(3)H showed a range of labeled PFC. The majority were in the plasmacytic category, including some mature plasma cells. The data from the experiments with in vivo labeling suggest a direct differentiation from antibody-synthesizing lymphocytes to plasma cells. Further, the in vivo experiments indicated that differentiation from nascent lymphocyte to plasma cell could be essentially completed within 1 day.