Anti-DNP Hemolytic Piaques by Mouse Spieeiv Celis in Diffusion Chambers

Abstract

Spleen cells frcan mice immunized with dinitrophenylated keyhole limpet hemocyanin (DNP-KLH) were restimulated with DNP-KLH in vitro, enclosed in cell-impermeable diffusion chambers and implanted in the peritoneal cavity of irradiated recipient mice. At various intervals, the cells were recwered from the chambers and DKP-specific plaque forming cells (PFC) were enumerated. The numbers of nucleated cells in the chambers declined rapidly during the first day after implantstion then increased between days 3 and 6 and remained at between 30 and 40% of the initial number from day 6 to day 10. Doses of DNF-KLH of 5 to 500 ng induced maximal numbers of PFC, while fewer PEC were obtained Kith lower and higher doses. The numbers of PFC/10⁶ cells declined during the first 2 days of culture in diffusion chambers, then increased explosively between days 3 and 4 and reached a peak on day 5 through 8. At peak response, 5% of the nucleated cells were PFC and in one case the proportion of PFC was 12%. Varying the time between primary impaunization in vivo and secondary inunmization in vitro from 2 to 11 weeks had no effect on the kinetics of the plaque response.