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Publisher Summary This chapter describes a method that helps researchers determine lymphocyte–macrophage interactions linked to delayed hypersensitivity. The method involves sensitizing guinea pigs by inoculation of 0.5 ml complete Freund's adjuvant with or without additional heterologous antigen, equally among the 4 footpads. A 2.5 ml glass Luer–Lock syringe with a 22 gauge needle is usually used. The degree of hypersensitivity is determined by skin testing the animals on the shaved flank with an appropriate amount of antigen. The reactions are observed and recorded 4 and 24 hours later. Five to seven days after the injection of oil and after preparation of the cell culture medium, peritoneal exudates cells are harvested. The animals are anesthetized and sacrificed. They are then pinned to an animal board and the hair on the ventral surface is shaved with an electric clipper. The hardened agar-containing medium, with or without specific antigen, is then inoculated by careful placement of small drops of concentrated cells on the agar surface with a capillary pipette. |
