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The World of Protozoa, Rotifera, Nematoda and Oligochaeta

Ref ID : 4360

Krista Fischer-Stenger, Guy A. Cabral, and Francine Marciano-Cabral; Separation of Soluble Amoebicidal and Tumoricidal Activity of Activated Macrophages. J.Protozool. 39(1):235-241, 1992

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Macrophage-conditioned medium (M(phi)CM) prepared from mouse peritoneal macrophages activated in vivo with bacillus Calmette-Guerin (BCG) or propionibacterium acnes and triggered with lipopolysaccharide in vitro contained tumoricidal and amoebicidal activity. The murine fibroblast cell line L929 was used as the indicator of tumoricidal activity and Naegleria fowleri amoeba was used to detect amoebicidal activity in M(phi)CM. The protease inhibitor, soybean trypsin inhibitor, decreased tumoricidal activity but had little effect on amoebicidal activity in M(phi)CM. Anti-TNF(alpha) antiserum inhibited tumoricidal activity in M(phi)CM. The antiserum reduced amoebicidal activity in BCG-activated M(phi)CM but had no effect on amoebicidal activity in P. acnes-activated M(phi)CM. Recombinant TNF(alpha), rIL-1(alpha), or rIL-1(beta) independently did not affect cytolysis of amoebae. Also, rTNF(alpha) had no effect on the growth of amoebae. Preparative flat-bed electrofocusing of BCG-activated M(phi)CM yielded fractions that exhibited different amoebicidal and tumoricidal activity profiles. Three domains of activity were analyzed (acidic, neutral, and basic). Anti-TNF(alpha) antiserum eliminated tumoricidal activity, but not amoebicidal activity, in fractions from the acidic domain. A combination of anti-TNF(alpha) and anti-IL-1(alpha) antisera failed to eliminate amoebicidal activity in fractions from the basic domain. These results indicate that different factors are responsible for macrophages amoebicidal and tumoricidal activity. The amoebicidal factors in M(phi)CM affected cytolysis of several species of amoebae.