Inflammatory immune response of canine trophoblastic cells from primary culture to infection with Brucella canis

ANDREA G. FERNANDEZ; FERRERO, MC; HIELPOS, MS; FOSSATI, CA; BALDI, PC

Mar del Plata, Buenos Aires

Congreso; VI Latin American Symposium on Maternal-Fetal Interaction and Placenta and V Latin American Symposium on Reproductive Immunology Meeting 2015; 2015

SLIMP & LASRI

Brucella canis is an important cause of reproductive complications in dogs, including abortion and perinatal death of newborns. However, the pathogenic mechanisms by which this bacterium causes abortion have not been described so far. Histological studies point to trophoblasts as target cells of B. canis.Objectives: 1) to confirm B. canis ability to infect and replicate inside canine trophoblastic cells; 2) to analyze the inflammatory immune response of infected trophoblasts.Methods: canine trophoblastic cells were isolated from term placenta obtained by C-section of healthy female dogs and were characterized by immune fluorescence with anti-cytokeratin 7 (ck7) antibody. 5x104 cells/well were infected with B. canis (clinical isolate) at MOI 250 for 48 h. After treatment with gentamicin, cells were lysed and plated onto bacteriological agar for CFU determination at 2, 24 and 48 h post infection (p.i.). Culture supernatants were collected to evaluate cytotoxicity (LDH activity) and proinflammatory cytokines (by ELISA).Results: Isolated canine trophoblasts were positive for ck7 staining. Replication kinetics showed a significant decrease (p