Macrophage activating factors produced in the course of murine tularemia: effect on multiplication of microbes
Kovarova, H.; Marcela, A.; Stulik, J.
Archivum Immunologiae et Therapiae Experimentalis 40(3-4): 183-190
1992
ISSN/ISBN: 0004-069X PMID: 1300982 Document Number: 400645
Primary Francisella tularensis infection in mice induces the production of macrophage activating factors (MAFs) by spleen cells. The stimulation of macrophage cytolytic activity (MAF-c) and hydrogen peroxide production (MAF-H-2O-2) dominates between days 7 and 10 in the course of tularemia. Three various pools of active fractions (10-11, 14-15, 25-28) were fractionated by two-step chromatography. Typical for 10-11 and 14-15 is MAF-c activity whereas in 25-28 prevails MAF-H-2O-2. Initial concentrated supernatant (day 7 of infection) and individual fractions have been used to raise antibodies KI (anti 10-11) and KII (anti 14-15). Neutralization reactions with specific antibodies indicate the presence of tumor necrosis factor alpha (TNF-alpha) in 14-15 (44% inhibitable), interferon gamma (IFN-gamma) and interleukin 2 (IL 2) in 25-28 (65% and 30% neutralization, respectively). Utilizing KI and KII, 99% and 90% inhibition of cytolytic activity is reached in 10-11 and 14-15, respectively, in spite of non-specific cross reaction. Western blot analysis of proteins in supernatant on day 7 detects, besides TNF-alpha, further protein bands (13, 15.5, 52 and 72 kDa) that seems to be associated with macrophage activation. Significantly protective effect against in vivo multiplication of tularemic microbes indicates a certain role of TNF-alpha, however, cooperation of other molecules is worth to be taken into consideration.