BRUCELLA INFECTION AFFECTS FUNCTIONALITY OF DECIDUA STROMAL CELLS AND TROPHOBLASTS

ZAVATTIERI LUCÍA; MACCHI ROSARIO; FLORENCIA MUÑOZ GONZALEZ; ANDREA CANELLADA; FERRERO MARIANA CRISTINA; BALDI PABLO C

San Luis

Congreso; LXXI Reunion Anual de la Sociedad Argentina de Inmunología; 2023

Sociedad Argentina de Inmunología

BRUCELLA INFECTION AFFECTS FUNCTIONALITY OF DECIDUA STROMAL CELLS AND TROPHOBLASTS Lucía Zavattieri1,2, Rosario Macchi1,2, Florencia Muñoz González, Andrea M. Canellada1,2 , Mariana C. Ferrero1,2 , Pablo C. Baldi1,2 1 IDEHU (CONICET-UBA), 2Facultad de Farmacia y Bioquímica UBA, Buenos Aires, Argentina The ability of trophoblasts to migrate, form tubes and invade the decidua is essential for a successful pregnancy. These processes are modulated by cytokines produced by decidual cells. Brucella infections are associated with reproductive complications in humans and animals. We evaluated if Brucella can impair functions of decidual cells and trophoblasts that are relevant for a successful pregnancy. The effect of Brucella infection on trophoblast migration was evaluated with the scratch test using the human trophoblastic cell line Swan71. Wound closure was significantly diminished in trophoblasts infected with B. abortus (Multiplicity of Infection 50: 45%; MI 500: 36% vs. 83% in uninfected controls), and similar results were observed with Brucella suis and Brucella melitensis infections. Human endometrial stromal cells (THESC) were infected with different Brucella species (B. abortus, B. suis, or B. melitensis) before or after decidualization with medroxyprogesterone and cAMP, and conditioned media (CM) were used to stimulate Swan71 cells during the scratch test. Wound closure was reduced in Swan71 cells treated with CM from B. abortus-infected decidualized THESC as compared to uninfected CM (20% vs. 86%). Similarly, wound closure was inhibited by CM obtained from THESC cells infected before decidualization (8.5% vs. 72.8%). Comparable results were observed with the other strains. Inflammatory signals are known to affect or impact trophoblast functions. Trophoblast migration (wound closure) improved when CM from B. abortus-infected THESC at different MI was treated with neutralizing antibodies (Ab) against CXCL8 or CCL2 chemokines before addition to the scratch test. Incubation with CM plus anti-IL-8 antibody increased wound closure by 59 and 45 % for MI 50 and 500 respectively vs. CM untreated, while, anti-CCL2 antibody treatment increased 55% and 52% for MI 50 and 500 respectively. Moreover, trophoblast tube formation assay was more successful in uninfected controls (NI) being reduced in the presence of infected THESC CM, as revealed by the number of master junctions (NI: 88; MI 50: 33; MI 500: 19), master segments (NI:152; MI 50: 28; MI 500: 46.5 and meshes (NI:56; MOI 50: 16; MOI 50: 20; MOI 500:13.5). In addition, when THESC were infected, the attachment area of trophoblast spheroids (invasion) was affected (NI:0.98; MOI 50: 0.71; MOI 250: 0.53; MOI 500: 0.47). Overall, the normal function of trophoblasts and decidual cells, and cross-talk between them, can be affected by Brucella infection