PEDV transmission assays and conjugate formation
PEDV-loaded RBCs were labeled with
CM-Dil Dye, washed extensively to
remove unbound virus and dye, and then co-cultured with PBMCs (RBCs:
PBMCs = 1:1) for 1 h or 6 h. For detection of PEDV transmission, after
co-culture, RBCs were removed by ACK Lysis Buffer and PBMCs were
collected to detect by
flow
cytometry and Western blot. For observation the conjugated formation,
CD3+ T cells were purified from PBMCs as described (Li et al., 2018).
CD3+ T cells were
then co-cultured with virus-loaded RBCs and then centrifuged at 200×g
for 5 min. The mixtures of RBCs and PBMCs were detected by flow
cytometry and TEM observation.