Intracellular bacterial replication and cytokine secretion in canine placental explants infected ex-vivo with B. canis.
Explants were obtained from 3 term placentas collected by C-section of healthy dogs, and were infected with 1x108 CFU of B. canis for 2 h. The explants were washed with PBS and extracellular bacteria were killed by the addition of antibiotics. (A) Explants homogenates were obtained at 2, 24 and 48 h p.i. and plated on TSA to count intracellular CFU. (B) Culture supernatants were collected at 24 and 48 h p.i. to measure TNF-α, RANTES (CCL5), IL-8 and IL-6 secretion. Values are means ± SD of duplicate measurements from three independent experiments. In panel A, asterisks indicate significant differences as compared to 2 h. p.i. (** p<0.01, Mann-Whitney test). In panel B, asterisks indicate significant differences as compared to uninfected (control) cells (* p<0.05; ** p<0.01, Student᾽s t test).
