Journal: mSphere

Article Title: Term Human Placental Trophoblasts Express SARS-CoV-2 Entry Factors ACE2, TMPRSS2, and Furin

doi: 10.1128/mSphere.00250-21

Figure Lengend Snippet: The ACE2-dependent entry of SARS-CoV-2 pseudovirus into cultured PHT cells. (A) PHT (five donors) and 293T-ACE2 cells inoculated with SARS-CoV-2 protein pseudotyped lentiviruses containing SMNE proteins (spike, membrane, nucleocapsid, and envelope) or MNE proteins, as detailed in Materials and Methods. Cells, harvested on day 2 postinoculation with the virus, were washed three times with PBS and trypsinized to remove adherent virus, and the cell pellet was lysed to release intracellular p24. Shown is p24 content (in picograms per milliliter) in cell lysates. *, P < 0.01 (paired t test). (Inset) 293T cells expressing ACE2, assessed at 2 days or 5 days ( n = 2 for each) and serving as a positive control. (B) PHT were preincubated or not with 20 μg/ml of anti-ACE-2 or anti-DC-SIGN antibody (Ab) and then inoculated with pvSARS-CoV-2 S+. The PHT cells were also inoculated with pseudovirus lacking S protein (pvS-) or with inactivated HIV-1 as negative controls. Cells were harvested on day 2 postinoculation, and p24 content (in picograms per milliliter) was measured in cell lysates, as detailed in Materials and Methods. *, P < 0.05 (ANOVA and Tukey test, n = 5).

Article Snippet: For antibody inhibition experiments, PHT cells were preexposed to 20 μg/ml anti-ACE2 (catalog no. AF933; R&D Systems, Minneapolis, MN ) or anti-DC-SIGN (R&D catalog no. MAB161) for 30 min at 37°C before inoculation with the viruses ( , ).

Techniques: Cell Culture, Expressing, Positive Control