1. An In Vitro Model for Toxin-Mediated Vascular Leak Syndrome: Ricin Toxin A Chain Increases the Permeability of Human Endothelial Cell Monolayers
by Alan L. Lindstrom, Stanley L. Erlandsen, John H. Kersey, and Christopher A. Pennell
Blood
Volume 90(6):
September 15, 1997
©1997 by American Society of Hematology

2. Schematic representation of a filtration unit.
Schematic representation of a filtration unit. This is one of 12 units arranged in a 2 by 6 array in the TEFF apparatus.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

3. Thrombin-induced permeability.
Thrombin-induced permeability. HUV-EC-C monolayers cultured on collagen IV-coated microporous cell culture inserts were either left untreated (n = 3; ▪) or exposed to 5 U/mL thrombin (n = 3; ▨) for 15 minutes. Monolayer permeability was measured by TEFF and was expressed as the mean fold change in fluid flux ± standard deviation. The mean pretreatment fluid flux values for untreated and thrombin-treated monolayers were 84 and 141 (μL/min)/cm2, respectively. Statistically significant differences between the mean responses of treated and time-matched control monolayers are depicted as follows: **P < .01.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

4. dgRTA-induced permeability.
dgRTA-induced permeability. HUV-EC-C monolayers cultured on collagen IV-coated microporous cell culture inserts were either left untreated (n = 5; ○) or were exposed to 10−7 mol/L (3.0 μg/mL) dgRTA (n = 4; ♦). Monolayer permeability was measured by TEFF and is expressed as the mean ± standard deviation. Data from two separate experiments are presented here. Statistically significant differences between the mean responses of treated and time-matched control monolayers are depicted as follows: *P < .05; **P < .01.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

5. Alan L. Lindstrom et al. Blood 1997;90:2323-2334
©1997 by American Society of Hematology

6. Alan L. Lindstrom et al. Blood 1997;90:2323-2334
©1997 by American Society of Hematology

7. DA7-induced permeability.
DA7-induced permeability. HUV-EC-C monolayer cultures and permeability measurements were performed as in Fig 3. HUV-EC-C monolayers were either left untreated (n = 4; ○) or were exposed to 10−8 mol/L DA7 (n = 6; •). Monolayer permeability was measured by TEFF and is expressed as the mean ± standard deviation. Statistically significant differences between the mean responses of treated and time-matched control monolayers are depicted as follows: *P < .05.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

8. Effect of dgRTA on the permeability of HUVE and HUV-EC-C monolayers.
Effect of dgRTA on the permeability of HUVE and HUV-EC-C monolayers. HUV-EC-C and HUVE monolayers were cultured and permeability was assessed as in Fig 3. Passage 19 HUV-EC-C (circles) or pooled passage 4 HUVE monolayers (diamonds) were either left untreated (open symbols) or treated with dgRTA (10−8 mol/L; 0.3 μg/mL; solid symbols). To facilitate comparing results derived from the different cell types, the data are expressed as the mean fold change in fluid flux relative to pretreatment fluid flux. The mean pretreatment fluid flux values for HUV-EC-C and HUVE cells in this experiment were 4.2 and 10.5 (μL/min)/cm2, respectively.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

9. dgRTA-induced morphologic changes.
dgRTA-induced morphologic changes. HUV-EC-C monolayers were cultured on the collagen IV-coated microporous cell culture inserts and treated as they were in the TEFF assays. SEM images represent (A) naked microporous membranes, (B) untreated control monolayers, (C) monolayers treated with 10−7 mol/L dgRTA for 96 hours, or (D) monolayers treated with 1 μg/mL cytochalasin D for 15 minutes. All photomicrographs were taken at 500× magnification using a tilt angle of 45°. The bar in (A) represents 50 μm; the original magnifications for (B) through (D) are the same as (A). Arrows indicate exposed pores and arrowheads indicate pores covered by endothelial cells.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

10. Temporal effect of dgRTA on exposed porosity.
Temporal effect of dgRTA on exposed porosity. Treatment with 10−7 mol/L dgRTA induced intercellular gaps in the HUV-EC-C monolayer that exposed pores in the support membrane. An area of ≥600 μm2 per sample was examined and the number of exposed pores were determined. Data are expressed as the mean ± standard deviation. A significance level of P < .05 is indicated by an asterisk.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

11. Comparison of dgRTA-, rRTA-, and rRTA-E177D–induced permeability.
Comparison of dgRTA-, rRTA-, and rRTA-E177D–induced permeability. HUV-EC-C monolayer cultures and permeability measurements were performed as in Fig 3. Monolayers were either left untreated (n = 5; ○) or exposed to 10−7 mol/L dgRTA (n = 4; ♦), rRTA (n = 5; ⋄), or rRTA-E177D (n = 4; □). Permeability is expressed as the mean fluid flux ± standard deviation. Data are combined from two separate experiments. Statistically significant differences between the mean responses of treated and time-matched control monolayers are depicted as follows: *P < .05; **P < .01.
Alan L. Lindstrom et al. Blood 1997;90:
©1997 by American Society of Hematology

12. Alan L. Lindstrom et al. Blood 1997;90:2323-2334
©1997 by American Society of Hematology

13. Alan L. Lindstrom et al. Blood 1997;90:2323-2334
©1997 by American Society of Hematology