Separation of Soluble Amoebicidal and Tumoricidal Activity of Activated Macrophages |
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Authors: | KRISTA FISCHER-STENGER GUY A CABRAL FRANCINE MARCIANO-CABRAL |
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Institution: | Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, Virginia 23298-0678 |
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Abstract: | ABSTRACT. Macrophage-conditioned medium (MøCM) prepared from mouse peritoneal macrophages activated in vivo with bacillus Calmette-Guérin (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 MøCM. The protease inhibitor, soybean trypsin inhibitor, decreased tumoricidal activity but had little effect on amoebicidal activity in MøCM. Anti-TNF α antiserum inhibited tumoricidal activity in MøCM. The antiserum reduced amoebicidal activity in BCG-activated MøCM but had no effect on amoebicidal activity in P. acnes -activated MøCM. Recombinant TNF α , rIL-1 α , or rIL-1 β independently did not affect cytolysis of amoebae. Also, rTNF α had no effect on the growth of amoebae. Preparative flat-bed electrofocusing of BCG-activated MøCM yielded fractions that exhibited different amoebicidal and tumoricidal activity profiles. Three domains of activity were analyzed (acidic, neutral, and basic). Anti-TNF α antiserum eliminated tumoricidal activity, but not amoebicidal activity, in fractions from the acidic domain. A combination of anti-TNF α and anti-IL-1 α antisera failed to eliminate amoebicidal activity in fractions from the basic domain. These results indicate that different factors are responsible for macrophage amoebicidal and tumoricidal activity. The amoebicidal factors in MøCM affected cytolysis of several species of amoebae. |
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Keywords: | Amoeba interleukin-1α interleukin-1β Naegleria fowleri protease inhibitor protozoa tumor necrosis factorα |
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