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A Cell-to-cell transmission > (Relative, %) C Infection (RLU x10³) E Infection (RLU x104) G 1.51 1.0 0.5- 0.0- 2 + 3 NO SARS-CoV (PV) SARS-CoV-2 (PV) 0 0.2 0.5 1.5 0 0.2 0.5 1.5 293T to H520 (SARS-CoV-2 (PV)) Day 293T to PBMC (SARS-CoV-2 (PV)) 4 Day 20 h →Cell-to-cell 4 Cell-free 1.51 Cell-to-cell +Cell-free 1.0- 0.5- mTomato-Red 2 5.24% 12.3% 1.18% 4h 3 73.2% 6.17% 9.32% 7.45% 0.507% 1.31% 71.7% 14.7% 0.713% ACE2 (ug) B Cell-free infection (Relative, %) D Infection (RLU x10³) F 85.0T Infection (RLU x104) 2.51 2.0- 1.5- 1.0- 0.5- 0.0- 801 60- Cell-to-cell (NCI-H520) Cell-free (NCI-H520) SARS-CoV (PV) SARS-CoV-2 (PV) 0 0.2 0.5 1.5 0 0.2 0.5 1.5 293T to H520 (SARS-CoV (PV)) →Cell-to-cell +Cell-free 4 Day 293T to PBMC (SARS-CoV (PV)) →-Cell-to-cell 4 Day SARS-CoV-2 WA1 (authentic) Cell-free (co-culture) Cell-to-cell www 20+Cell-free 0 H Infection (Q3, %) 201 15- 10- 0. ACE2 (ug)

A Cell-to-cell transmission > (Relative, %) C Infection (RLU x10³) E Infection (RLU x104) G 1.51 1.0 0.5- 0.0- 2 + 3 NO SARS-CoV (PV) SARS-CoV-2 (PV) 0 0.2 0.5 1.5 0 0.2 0.5 1.5 293T to H520 (SARS-CoV-2 (PV)) Day 293T to PBMC (SARS-CoV-2 (PV)) 4 Day 20 h →Cell-to-cell 4 Cell-free 1.51 Cell-to-cell +Cell-free 1.0- 0.5- mTomato-Red 2 5.24% 12.3% 1.18% 4h 3 73.2% 6.17% 9.32% 7.45% 0.507% 1.31% 71.7% 14.7% 0.713% ACE2 (ug) B Cell-free infection (Relative, %) D Infection (RLU x10³) F 85.0T Infection (RLU x104) 2.51 2.0- 1.5- 1.0- 0.5- 0.0- 801 60- Cell-to-cell (NCI-H520) Cell-free (NCI-H520) SARS-CoV (PV) SARS-CoV-2 (PV) 0 0.2 0.5 1.5 0 0.2 0.5 1.5 293T to H520 (SARS-CoV (PV)) →Cell-to-cell +Cell-free 4 Day 293T to PBMC (SARS-CoV (PV)) →-Cell-to-cell 4 Day SARS-CoV-2 WA1 (authentic) Cell-free (co-culture) Cell-to-cell www 20+Cell-free 0 H Infection (Q3, %) 201 15- 10- 0. ACE2 (ug)

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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A Cell-to-cell transmission > (Relative, %) C Infection (RLU x10³) E 1.5 1.0 0.5- 0.0- Infection (RLU x10¹) G 5- 2 + 3 N 4 Cell-free 1.51 Cell-to-cell +Cell-free 1.0- 0.5- 0 H mTomato-Red 2 0 0.2 0.5 1.5 293T to H520 (SARS-CoV-2 (PV)) →Cell-to-cell 2 4 Day 293T to PBMC (SARS-CoV-2 (PV)) 4 Day 20 h 5.24% I SARS-CoV (PV) SARS-CoV-2 (PV) 4h ACE2 0 0.2 0.5 1.5 (pg) B 71.7% Cell-free infection (Relative, %) D Infection (RLU x10³) F 3 73.2% 6.17% 201 Cell-to-cell (NCI-H520) 15- 14.7% 12.3% 1.18% 9.32% 7.45% 0.507% 1.31% 10- 0.713% 5- Cell-free (NCI-H520) 0. 198.0% 0.713% 0.338% 97.3% SARS-CoV-2 N (FITC) Fig. 4. ACE2 enhances cell-to-cell transmission but is not absolutely required. (A and B) Cell-to-cell and cell-free infection were performed as described for Figs. 1 and 3 except that target cells were 293T transfected with different amounts of a plasmid encoding ACE2. Relative rates of cell-to-cell transmis- sion and cell-free infection were calculated by setting the values of 0.5 µg ACE2 to 1.0 (A and B, n = 3). (C-F) Experiments were carried out as described for Figs. 1 and 3 except that target cells were H520 and human PBMCs (n = 3 for each). (G) Vero-ACE2-TMPRSS2-mTomato (Red) cells were infected with MOI = 0.01 of authentic SARS-CoV-2 WT (USA-WA1/2020) for 18 h, followed by coculturing of the donor NCI-H520 cells for another 4 h or 20 h. Cells were fixed and stained with an anti-SARS-CoV-2 N protein for flow cytometric analysis. The same number of NCI-H520 cells were infected with the WT SARS-CoV-2 harvested from the SARS-CoV-2-infected donor Vero-ACE2-TMPRSS2-mTomato cells for 4 h or 20 h for cell-free infection and analyzed by flow cytometry. (H) Results of Q3 quadrant analyses were plotted. ***P<0.001. PV, pseudotyped virus. 2.51 2.0- 1.5- 1.0- 0.5- 0.0- SARS-CoV (PV) SARS-CoV-2 (PV) I 푸 ACE2 0 0.2 0.5 1.5 0 0.2 0.5 1.5 (ug) 293T to H520 (SARS-CoV (PV)) 2 4 Day 293T to PBMC (SARS-CoV (PV)) …‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒ 4 Day SARS-CoV-2 WA1 (authentic) Cell-free (co-culture) Cell-to-cell *** Infection (RLU x104) 801 →Cell-to-cell 60+Cell-free 40 20- 5.0T 2.5 251 Cell-to-cell 20 →→Cell-free 15 10- F 5- 2.01 드 1.01 0- H Infection (Q3, %) 2 20 h 4h 20 h MICROBIOLOGY
Transcribed Image Text:A Cell-to-cell transmission > (Relative, %) C Infection (RLU x10³) E 1.5 1.0 0.5- 0.0- Infection (RLU x10¹) G 5- 2 + 3 N 4 Cell-free 1.51 Cell-to-cell +Cell-free 1.0- 0.5- 0 H mTomato-Red 2 0 0.2 0.5 1.5 293T to H520 (SARS-CoV-2 (PV)) →Cell-to-cell 2 4 Day 293T to PBMC (SARS-CoV-2 (PV)) 4 Day 20 h 5.24% I SARS-CoV (PV) SARS-CoV-2 (PV) 4h ACE2 0 0.2 0.5 1.5 (pg) B 71.7% Cell-free infection (Relative, %) D Infection (RLU x10³) F 3 73.2% 6.17% 201 Cell-to-cell (NCI-H520) 15- 14.7% 12.3% 1.18% 9.32% 7.45% 0.507% 1.31% 10- 0.713% 5- Cell-free (NCI-H520) 0. 198.0% 0.713% 0.338% 97.3% SARS-CoV-2 N (FITC) Fig. 4. ACE2 enhances cell-to-cell transmission but is not absolutely required. (A and B) Cell-to-cell and cell-free infection were performed as described for Figs. 1 and 3 except that target cells were 293T transfected with different amounts of a plasmid encoding ACE2. Relative rates of cell-to-cell transmis- sion and cell-free infection were calculated by setting the values of 0.5 µg ACE2 to 1.0 (A and B, n = 3). (C-F) Experiments were carried out as described for Figs. 1 and 3 except that target cells were H520 and human PBMCs (n = 3 for each). (G) Vero-ACE2-TMPRSS2-mTomato (Red) cells were infected with MOI = 0.01 of authentic SARS-CoV-2 WT (USA-WA1/2020) for 18 h, followed by coculturing of the donor NCI-H520 cells for another 4 h or 20 h. Cells were fixed and stained with an anti-SARS-CoV-2 N protein for flow cytometric analysis. The same number of NCI-H520 cells were infected with the WT SARS-CoV-2 harvested from the SARS-CoV-2-infected donor Vero-ACE2-TMPRSS2-mTomato cells for 4 h or 20 h for cell-free infection and analyzed by flow cytometry. (H) Results of Q3 quadrant analyses were plotted. ***P<0.001. PV, pseudotyped virus. 2.51 2.0- 1.5- 1.0- 0.5- 0.0- SARS-CoV (PV) SARS-CoV-2 (PV) I 푸 ACE2 0 0.2 0.5 1.5 0 0.2 0.5 1.5 (ug) 293T to H520 (SARS-CoV (PV)) 2 4 Day 293T to PBMC (SARS-CoV (PV)) …‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒‒ 4 Day SARS-CoV-2 WA1 (authentic) Cell-free (co-culture) Cell-to-cell *** Infection (RLU x104) 801 →Cell-to-cell 60+Cell-free 40 20- 5.0T 2.5 251 Cell-to-cell 20 →→Cell-free 15 10- F 5- 2.01 드 1.01 0- H Infection (Q3, %) 2 20 h 4h 20 h MICROBIOLOGY
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