Figure 3.

Trophoblast cells instruct pNKs to exhibit dNK cell-like phenotypes via interactions between Tim-3 and Gal-9. (a) FCM was performed to analyze intracellular expression of Gal-9 in trophoblasts. (b) An ELISA assay was used to assess the secretion of Gal-9 from human primary trophoblasts cultured for different periods of time. (c–h) Purified pNK cells from the first trimester were cultured with trophoblasts in the presence or absence of anti-Tim-3 neutralizing Abs for 72 h. FCM analysis was used to determine the levels of (c, d) Th1-type cytokines, (e, f) Th2-type cytokines and (g, h) cytotoxicity of pNK cells. The results are representative of 9 independent experiments. Data are shown as the mean±s.e.m. *P<0.05; **P<0.01; ***P<0.001. FCM, flow cytometry; pNK, peripheral NK cells; Tro, trophoblast; α-Tim-3, anti-Tim-3 neutralizing antibodies.