This study aims to assess whether soluble HLA-G (sHLA-G) is involved in apoptosis of decidual natural killer (dNK) cells following Toxoplasma gondii infection. dNK cells or NK-92 cells were infected with T. gondii and co-cultured with trophoblast cells or BeWo cells. Infected co-cultured cells were treated without or with sHLA-G neutralizing antibody. Uninfected co-cultured cells were used as controls. Apoptosis of dNK cells were analyzed by flow cytometry and confocal microscope. Real-time PCR and Western blot were used to determine caspase 3 and caspase 8 expression. sHLA-G in supernatant were measured by enzyme-linked immunosorbent assay (ELISA). In infection groups, sHLA-G was increased, while dNK apoptosis proteins caspase 3 and caspase 8 were up-regulated, but significantly decreased in the presence of sHLA-G neutralizing antibody compared to controls. Under the situation of T. gondii-infected dNK cells co-cultured with trophoblast cells, the up-regulation of sHLA-G could induce dNK cells apoptosis which ultimately may contribute to the abnormal pregnancy outcomes with T. gondii infection.