The production of anti-guinea pig lymphokine antibody

Abstract

Rabbit antiserum to lymphokine-containing supernatant fluids has been prepared by means of a two-stage immunization procedure. Migration inhibiting factor (MIF)-containing and control supernatants were obtained from guinea pig lymphocyte cultures. Both were fractionated on Sephadex G-100. Fractions of control supernatants corresponding to active fractions from the MIF-rich material were used to immunize rabbits. The anti-"control" antibody so obtained was conjugated to agarose beads to prepare an immunoadsorbent column. The MIF-rich Sephadex fractions were passed through this column to remove materials corresponding to those found in control supernatants and which could react with the anti-control antibody. The eluted MIF-rich fraction was concentrated and used to immunize a second group of rabbits. The anti-"lymphokine" antiserum so obtained was studied for its effect in vitro and in vivo. It was found capable of suppressing the activity of MIF, macrophage chemotactic factor, and skin reactive factor. It could also suppress delayed hypersensitivity skin reactions in actively immunized guinea pigs.