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.
| Original language | English |
|---|---|
| Pages (from-to) | 11-8 |
| Number of pages | 8 |
| Journal | Journal of Immunological Methods |
| Volume | 127 |
| Issue number | 1 |
| Publication status | Published - 20 Feb 1990 |
Keywords
- Animals
- Cells, Cultured
- Colorimetry
- Cytotoxicity, Immunologic
- Female
- Interferon-gamma
- Leishmania donovani
- Lipopolysaccharides
- Macrophage Activation
- Mice
- Mice, Inbred C57BL
- Recombinant Proteins
- Sodium Dodecyl Sulfate