We found that LPG induced opposing effects on the PMA-induced oxidative burst of macrophages from both mouse strains. Whereas in macrophages
of BALB/c mice, LPG inhibited the oxidative burst by 11·33%, compared with control values (P < 0·002), in C57BL/6 macrophages, LPG enhanced the oxidative burst by 13·7% (P < 0·017) over the controls not incubated with LPG. When the macrophages were pre-incubated with the PKCα inhibitor Gö6976, either alone or in combination with LPG, the oxidative burst of macrophages of both mouse strains was inhibited in relation to the macrophages in the absence of Gö6976, albeit the degree of inhibition was higher in the BALB/c macrophages (P < 0·002) (Figure 3a). We also analysed the effect of L. mexicana promastigotes on the PMA-induced oxidative burst Acalabrutinib clinical trial of peritoneal macrophages obtained from both mouse strains. The assay was performed in the absence or presence of the PKCα inhibitor Gö6976. L. mexicana promastigotes significantly diminished the oxidative burst induced by PMA on macrophages of both mouse strains, yet the degree of inhibition was significantly higher in BALB/c macrophages (46·4%, P < 0·002) than in C57BL/6 macrophages (19·4%P < 0·00013) as compared with controls not incubated with the parasite. In the presence of Gö6976, the degree of inhibition exerted
by L. mexicana promastigotes on the oxidative burst of BALB/c macrophages was similar to that achieved without Gö6976. In ADP ribosylation factor C57BL/6 macrophages, Gö6976 was able to increase the Bcl-2 inhibitor degree of inhibition of the oxidative burst exerted by L. mexicana promastigotes, as compared with the oxidative burst in the absence of Gö6976 (P < 0·00013) (Figure 3b). The purity of peritoneal macrophages ranged between 80% and 85% (data not shown). The levels of oxidative burst in cells not stimulated
by PMA, but treated only with LPG or L. mexicana promastigotes, were also measured. In both cases, the levels of oxidative burst were below the detection level of the apparatus. To determine a possible correlation between PKCα activity and the effectiveness of burst oxidation with the intracellular survival of the parasite, peritoneal macrophages from both mouse strains were infected with L. mexicana promastigotes. The oxidative burst was then induced by PMA and the parasite survival was analysed. Results show that in C75BL/6 macrophages stimulated with PMA, only 66% (259 parasites/100 macrophages) of the parasites survived as compared with parasite survival in macrophages not stimulated with PMA (390 parasites/100 macrophages). In contrast, 92% (280 parasites/100 macrophages) survived in BALB/c macrophages as compared with parasite survival in nonstimulated macrophages (304 parasites/100 macrophages).