Fitness Cost of Litomosoides sigmodontis Filarial Infection in Mite Vectors; Implications of Infected Haematophagous Arthropod Excretory Products in Host-Vector Interactions
Figure 4
Excretory compounds from infected mites only stimulate KC and TNF-α production by APC. ((a) and (b)) 12-day infected or uninfected mites were washed twice in PBS 1X and groups of 100 mites were placed for 2 hours at 25°C or 37°C in 24-well plates with 200 μL of saline solution to stimulate salivary excretion. Supernatants were collected and the quantification of proteins was evaluated by the Bradford method. Uninfected individuals produced more proteins than uninfected at 25°C (Student’s -test, ) as well as at 37°C (Student -test, ); results are expressed as mean ± SEM. ((c) and (d)) Bone-marrow-derived APCs were seeded into 96-well plates (106/well) in 200 μL buffer (RPMI 1640, 10% FCS, 100 U P/S, 1% L-glutamine, 1% NEAA, 1% sodium pyruvate) and cultured for 24 hours (5% CO2; 37°C) with saline solution (vehicle), and supernatant from infected mites (Spnt Inf.), or supernatant from uninfected (Spnt Uninf.) mites; both excretory solutions were diluted in saline solution to 10 μg/mL total proteins. Levels of TNF-α and KC were measured by ELISA in the culture supernatant. A significant increase of TNF-α or KC (one-way ANOVA followed by Bonferroni post hoc test; and , resp.) was observed in culture supernatants after addition of infected mites excretory solution; results are expressed as mean ± SEM.
