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A modified colorimetric assay of macrophage activation for intracellular cytotoxicity against Leishmania parasites

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JournalJournal of Immunological Methods
DatePublished - 20 Feb 1990
Issue number1
Volume127
Number of pages8
Pages (from-to)11-8
Original languageEnglish

Abstract

An in vitro method is described which colorimetrically assesses the activation of macrophages for intracellular cytotoxicity against the obligate intracellular parasite Leishmania donovani. The assay system uses a highly purified macrophage population derived from 10-day murine bone marrow cultures. These were infected in vitro as a suspension culture with viable L. donovani amastigotes and then exposed to activating agents. After 48 h the intracellular parasites were released by SDS lysis of the macrophages. Surviving Leishmania organisms were quantitated by their conversion of the chromophore MTT. The sensitivity of this method was comparable with the established method of [3H]dThd incorporation. This assay system has been used to show that there is a dual signal requirement (recombinant interferon-gamma and bacterial endotoxin (LPS] for effective activation of macrophages for leishmanicidal activity.

    Research areas

  • Animals, Cells, Cultured, Colorimetry, Cytotoxicity, Immunologic, Female, Interferon-gamma, Leishmania donovani, Lipopolysaccharides, Macrophage Activation, Mice, Mice, Inbred C57BL, Recombinant Proteins, Sodium Dodecyl Sulfate

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