Abstract
Macrophage cytotoxin (MCT) can be induced from peritoneal exudate macrophage monolayers (PEMM) obtained from thioglycollate-treated mice, by exposure of PEMM to lipopolysaccharide (LPS) or to double-stranded polyinosinic: poly-cytidylic acid (poly-l:poly-C). MCT is highly labile even upon storage at 4 degrees C, and is irreversibly denatured by isolectricfocusing, polyacrylamide gel electrophoresis in the absence of sodium dodecyl sulfate (SDS), or by exposure to ethylene glycol. alpha-MCT [150,000 daltons (d)] has been highly purified (2,000- to 5,000-fold) from serum-free, PEMM supernatants by a scheme of concentration, molecular sieving on Ultrogel AcA 44, negative hydrophobic affinity chromatography on benzyl-agarose, and ion exchange chromatography on aminoethyl-agarose. The scheme results in high yield of MCT, in part because of the rapidity with which the labile toxin is manipulated due to the tandemization of the chromatographic steps.
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