Abstract
Studies of the mechanism of cell-mediated immune responses have been hampered by the lack of in vitro systems. While the inhibition of macrophage migration model had advanced our understanding of delayed hypersensitivity reacions, the technique is complex and cumbersome. We have recently developed a simple test ofr measuring cell-mediated immune responses which depends on the aggregation of peritoneal exudate cells (PEC) in suspension cultures.
PEC from guinea pigs with delayed hypersensitivity aggregate when the cells are cultured with the appropritate antigen. Diphtheria toxoid, PPD, egg albumin and keyhole limpet hemocyanin have been used in this system. Aggregation appears at 6 h and reaches a maximum at 24–25 h which is comparable to the time course of cutaneous delayed hypersensitivity reacions. This in vitro model also correlates with another cell-mediated response, allograft rejection. PEC from strain 13 guinea pigs previously grafted with skin from strain 2 animals aggregate in the presense of strain 2 cells. Aggregation does not occur with mixtures of PEC from guinea pigs of the same strain or different strains without prior grafting.
Peripheral blood leukocytes, spleen cells or lymph mode cells from sensitized animals cultured with antigen synthesize a factor which causes aggregation of PEC from nonsensitive guinea pigs. The aggregating cells are macrophages, and the titer of macrophage aggregation factor (MAF) is determined by serial dilution. Macrophage aggregation in vitro provides a simple semiquantitative test fro cell-mediated immune reacions and permits further analysis of the mechanism involved.
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Gotoff, S., Lolekha, S. Cell-mediated Immune Response in vitro. Pediatr Res 4, 437 (1970). https://doi.org/10.1203/00006450-197009000-00015
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DOI: https://doi.org/10.1203/00006450-197009000-00015