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. 1986 Apr;123(1):16–24.

Recombinant tumor necrosis factor and immune interferon act singly and in combination to reorganize human vascular endothelial cell monolayers.

A H Stolpen, E C Guinan, W Fiers, J S Pober
PMCID: PMC1888144  PMID: 2421578

Abstract

Human endothelial cells (HECs) in confluent primary culture reproduce the epithelioid organization of the vascular lining in situ. The addition of greater than or equal to 20 U/ml recombinant tumor necrosis factor (rTNF) or greater than or equal to 16 U/ml recombinant immune interferon (rIFN-gamma) causes HECs 1) to become elongated, 2) to overlap, 3) to rearrange their actin filaments, and 4) to lose their stainable fibronectin matrix. These changes develop over 72-96 hours and are reversible upon withdrawal of the mediator. In serially passaged HECs similar morphologic changes develop. Furthermore, rTNF and rIFN-gamma are each cytostatic for subconfluent passaged cultures. When added in combination, low concentrations of rTNF and rIFN-gamma act synergistically, whereas higher concentrations (eg, 100 U/ml rTNF and 200 U/ml rIFN-gamma) produce unique morphologic changes. Doubly treated primary HECs extend many long, overlapping, spinelike processes and expose the substratum. Doubly treated passaged HECs become extremely elongated and then shed in large numbers. These in vitro changes in endothelial cell morphology and behavior may underlie immunologically mediated vascular responses in vivo.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barak L. S., Yocum R. R., Nothnagel E. A., Webb W. W. Fluorescence staining of the actin cytoskeleton in living cells with 7-nitrobenz-2-oxa-1,3-diazole-phallacidin. Proc Natl Acad Sci U S A. 1980 Feb;77(2):980–984. doi: 10.1073/pnas.77.2.980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beutler B., Mahoney J., Le Trang N., Pekala P., Cerami A. Purification of cachectin, a lipoprotein lipase-suppressing hormone secreted by endotoxin-induced RAW 264.7 cells. J Exp Med. 1985 May 1;161(5):984–995. doi: 10.1084/jem.161.5.984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clark R. A., Dvorak H. F., Colvin R. B. Fibronectin in delayed-type hypersensitivity skin reactions: associations with vessel permeability and endothelial cell activation. J Immunol. 1981 Feb;126(2):787–793. [PubMed] [Google Scholar]
  4. Collins T., Lapierre L. A., Fiers W., Strominger J. L., Pober J. S. Recombinant human tumor necrosis factor increases mRNA levels and surface expression of HLA-A,B antigens in vascular endothelial cells and dermal fibroblasts in vitro. Proc Natl Acad Sci U S A. 1986 Jan;83(2):446–450. doi: 10.1073/pnas.83.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dumonde D. C., Pulley M. S., Paradinas F. J., Southcott B. M., O'Connell D., Robinson M. R., den Hollander F., Schuurs A. H. Histological features of skin reactions to human lymphoid cell line lymphokine in patients with advanced cancer. J Pathol. 1982 Dec;138(4):289–308. doi: 10.1002/path.1711380402. [DOI] [PubMed] [Google Scholar]
  6. Dvorak A. M., Mihm M. C., Jr, Dvorak H. F. Morphology of delayed-type hypersensitivity reactions in man. II. Ultrastructural alterations affecting the microvasculature and the tissue mast cells. Lab Invest. 1976 Feb;34(2):179–191. [PubMed] [Google Scholar]
  7. Fiers W., Remaut E., Devos R., Cheroutre H., Contreras R., Gheysen D., Degrave W., Stanssens P., Tavernier J., Taya Y. The human fibroblast and human immune interferon genes and their expression in homologous and heterologous cells. Philos Trans R Soc Lond B Biol Sci. 1982 Sep 24;299(1094):29–38. doi: 10.1098/rstb.1982.0103. [DOI] [PubMed] [Google Scholar]
  8. Freemont A. J., Ford W. L. Functional and morphological changes in post-capillary venules in relation to lymphocytic infiltration into BCG-induced granulomata in rat skin. J Pathol. 1985 Sep;147(1):1–12. doi: 10.1002/path.1711470102. [DOI] [PubMed] [Google Scholar]
  9. Gillis S., Ferm M. M., Ou W., Smith K. A. T cell growth factor: parameters of production and a quantitative microassay for activity. J Immunol. 1978 Jun;120(6):2027–2032. [PubMed] [Google Scholar]
  10. Groenewegen G., Buurman W. A., van der Linden C. J. Lymphokines induce changes in morphology and enhance motility of endothelial cells. Clin Immunol Immunopathol. 1985 Sep;36(3):378–385. doi: 10.1016/0090-1229(85)90058-3. [DOI] [PubMed] [Google Scholar]
  11. Jaffe E. A., Mosher D. F. Synthesis of fibronectin by cultured human endothelial cells. J Exp Med. 1978 Jun 1;147(6):1779–1791. doi: 10.1084/jem.147.6.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kull F. C., Jr, Jacobs S., Cuatrecasas P. Cellular receptor for 125I-labeled tumor necrosis factor: specific binding, affinity labeling, and relationship to sensitivity. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5756–5760. doi: 10.1073/pnas.82.17.5756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Montesano R., Mossaz A., Ryser J. E., Orci L., Vassalli P. Leukocyte interleukins induce cultured endothelial cells to produce a highly organized, glycosaminoglycan-rich pericellular matrix. J Cell Biol. 1984 Nov;99(5):1706–1715. doi: 10.1083/jcb.99.5.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Montesano R., Orci L., Vassalli P. Human endothelial cell cultures: phenotypic modulation by leukocyte interleukins. J Cell Physiol. 1985 Mar;122(3):424–434. doi: 10.1002/jcp.1041220313. [DOI] [PubMed] [Google Scholar]
  15. Paradinas F. J., Southcott B. M., O'Connell D., den Hollander F., Schuurs A. H., Pulley M. S., Dumonde D. C. Changes induced by local injection of human lymphoid cell lymphokine into dermal metastases of breast carcinoma: a light and electron microscopical study. J Pathol. 1982 Dec;138(4):309–323. doi: 10.1002/path.1711380403. [DOI] [PubMed] [Google Scholar]
  16. Pfeffer L. M., Wang E., Tamm I. Interferon effects on microfilament organization, cellular fibronectin distribution, and cell motility in human fibroblasts. J Cell Biol. 1980 Apr;85(1):9–17. doi: 10.1083/jcb.85.1.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pober J. S., Gimbrone M. A., Jr, Cotran R. S., Reiss C. S., Burakoff S. J., Fiers W., Ault K. A. Ia expression by vascular endothelium is inducible by activated T cells and by human gamma interferon. J Exp Med. 1983 Apr 1;157(4):1339–1353. doi: 10.1084/jem.157.4.1339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pober J. S., Gimbrone M. A., Jr Expression of Ia-like antigens by human vascular endothelial cells is inducible in vitro: demonstration by monoclonal antibody binding and immunoprecipitation. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6641–6645. doi: 10.1073/pnas.79.21.6641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Polverini P. J., Cotran R. S., Sholley M. M. Endothelial proliferation in the delayed hypersensitivity reaction: an autoradiographic study. J Immunol. 1977 Feb;118(2):529–532. [PubMed] [Google Scholar]
  20. Rubin B. Y., Anderson S. L., Sullivan S. A., Williamson B. D., Carswell E. A., Old L. J. High affinity binding of 125I-labeled human tumor necrosis factor (LuKII) to specific cell surface receptors. J Exp Med. 1985 Sep 1;162(3):1099–1104. doi: 10.1084/jem.162.3.1099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Scahill S. J., Devos R., Van der Heyden J., Fiers W. Expression and characterization of the product of a human immune interferon cDNA gene in Chinese hamster ovary cells. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4654–4658. doi: 10.1073/pnas.80.15.4654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shalaby M. R., Aggarwal B. B., Rinderknecht E., Svedersky L. P., Finkle B. S., Palladino M. A., Jr Activation of human polymorphonuclear neutrophil functions by interferon-gamma and tumor necrosis factors. J Immunol. 1985 Sep;135(3):2069–2073. [PubMed] [Google Scholar]
  23. Sobel R. A., Blanchette B. W., Bhan A. K., Colvin R. B. The immunopathology of experimental allergic encephalomyelitis. II. Endothelial cell Ia increases prior to inflammatory cell infiltration. J Immunol. 1984 May;132(5):2402–2407. [PubMed] [Google Scholar]
  24. Thornton S. C., Mueller S. N., Levine E. M. Human endothelial cells: use of heparin in cloning and long-term serial cultivation. Science. 1983 Nov 11;222(4624):623–625. doi: 10.1126/science.6635659. [DOI] [PubMed] [Google Scholar]
  25. Williamson B. D., Carswell E. A., Rubin B. Y., Prendergast J. S., Old L. J. Human tumor necrosis factor produced by human B-cell lines: synergistic cytotoxic interaction with human interferon. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5397–5401. doi: 10.1073/pnas.80.17.5397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Willms-Kretschmer K., Flax M. H., Cotran R. S. The fine structure of the vascular response in hapten-specific delayed hypersensitivity and contact dermatitis. Lab Invest. 1967 Sep;17(3):334–349. [PubMed] [Google Scholar]

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