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. 1985 Apr;75(4):1153–1161. doi: 10.1172/JCI111810

Characterization of the adhesion of the human monocytic cell line U937 to cultured endothelial cells.

P E DiCorleto, C A de la Motte
PMCID: PMC425439  PMID: 3988935

Abstract

Adhesion of blood-borne monocytes to the vascular endothelium is the first step in the infiltration of this leukocyte into the vessel wall or the interstitial space during inflammation. A significant role for the monocyte in both wound healing and atherogenesis is now well accepted. The molecular interactions involved in monocyte attachment to the endothelium are unknown. To study this phenomenon we have developed an in vitro system that uses the human monocytic tumor cell line U937 as a model for the blood-borne monocyte. 51Cr-labeled U937 cells were found to adhere with high affinity to cultured endothelial cells (ECs) from several sources. Much less binding was observed to either smooth muscle cells or fibroblasts from several species. Conditioned medium and cocultivation experiments ruled out the possibility that target cells could affect U937 cell binding by secretion of factors. Binding of U937 cells to porcine aortic ECs reached equilibrium after 30 min at 37 degrees C and 90 min at 4 degrees C with similar extent of binding at the two temperatures. Binding of U937 to the endothelium reached saturation at 9-12 U937 per porcine aortic EC (semi-confluent) with half-maximal binding at 1.5 X 10(6) U937 cells/ml. Bound cells dissociated with a half-life of 20 h at 37 degrees C. Adhesion of U937 cells was blocked by prior incubation of ECs with normal monocytes but not with platelets, lymphocytes, or neutrophils. Trypsin treatment or detergent solubilization of ECs inhibited U937 cell binding. A striking effect of EC density on monocytic cell adhesion was observed with bovine, rat, and porcine ECs. Confluent cultures of these cells exhibited negligible binding of U937, but when plated sparsely, the same cells were excellent targets for U937 cell adhesion. In addition, when confluent cultures of bovine aortic ECs were "wounded" with a cotton swab and then allowed to recover for 24 h at 37 degrees C, U937 cells were found to adhere most readily to the ECs migrating into the wound and neighboring the wound but not to ECs in the confluent monolayer away from the wound edge. These latter results may have implications for the focal adhesion of monocytes to the vessel wall in vivo.

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Selected References

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  1. Boogaerts M. A., Yamada O., Jacob H. S., Moldow C. F. Enhancement of granulocyte-endothelial cell adherence and granulocyte-induced cytotoxicity by platelet release products. Proc Natl Acad Sci U S A. 1982 Nov;79(22):7019–7023. doi: 10.1073/pnas.79.22.7019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bowen-Pope D. F., Ross R. Platelet-derived growth factor. II. Specific binding to cultured cells. J Biol Chem. 1982 May 10;257(9):5161–5171. [PubMed] [Google Scholar]
  3. Buchanan M. R., Vazquez M. J., Gimbrone M. A., Jr Arachidonic acid metabolism and the adhesion of human polymorphonuclear leukocytes to cultured vascular endothelial cells. Blood. 1983 Oct;62(4):889–895. [PubMed] [Google Scholar]
  4. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  5. Cookson F. B. The origin of foam cells in atherosclerosis. Br J Exp Pathol. 1971 Feb;52(1):62–69. [PMC free article] [PubMed] [Google Scholar]
  6. DiCorleto P. E., Bowen-Pope D. F. Cultured endothelial cells produce a platelet-derived growth factor-like protein. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1919–1923. doi: 10.1073/pnas.80.7.1919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Faggiotto A., Ross R., Harker L. Studies of hypercholesterolemia in the nonhuman primate. I. Changes that lead to fatty streak formation. Arteriosclerosis. 1984 Jul-Aug;4(4):323–340. doi: 10.1161/01.atv.4.4.323. [DOI] [PubMed] [Google Scholar]
  8. Fischer D. G., Pike M. C., Koren H. S., Snyderman R. Chemotactically responsive and nonresposive forms of a continuous human monocyte cell line. J Immunol. 1980 Jul;125(1):463–465. [PubMed] [Google Scholar]
  9. Fowler S. Characterization of foam cells in experimental atherosclerosis. Acta Med Scand Suppl. 1980;642:151–158. doi: 10.1111/j.0954-6820.1980.tb10947.x. [DOI] [PubMed] [Google Scholar]
  10. Fowler S., Shio H., Haley N. J. Characterization of lipid-laden aortic cells from cholesterol-fed rabbits. IV. Investigation of macrophage-like properties of aortic cell populations. Lab Invest. 1979 Oct;41(4):372–378. [PubMed] [Google Scholar]
  11. Gerrity R. G., Goss J. A., Soby L. Control of monocyte recruitment by chemotactic factor(s) in lesion-prone areas of swine aorta. Arteriosclerosis. 1985 Jan-Feb;5(1):55–66. doi: 10.1161/01.atv.5.1.55. [DOI] [PubMed] [Google Scholar]
  12. Gerrity R. G., Naito H. K. Ultrastructural identification of monocyte-derived foam cells in fatty streak lesions. Artery. 1980;8(3):208–214. [PubMed] [Google Scholar]
  13. Gerrity R. G., Schwartz C. J. Endothelial cell injury in early mild hypercholesterolemia. Prog Biochem Pharmacol. 1977;13:213–219. [PubMed] [Google Scholar]
  14. Gerrity R. G. The role of the monocyte in atherogenesis: I. Transition of blood-borne monocytes into foam cells in fatty lesions. Am J Pathol. 1981 May;103(2):181–190. [PMC free article] [PubMed] [Google Scholar]
  15. Gerrity R. G. The role of the monocyte in atherogenesis: II. Migration of foam cells from atherosclerotic lesions. Am J Pathol. 1981 May;103(2):191–200. [PMC free article] [PubMed] [Google Scholar]
  16. Glenn K. C., Ross R. Human monocyte-derived growth factor(s) for mesenchymal cells: activation of secretion by endotoxin and concanavalin A. Cell. 1981 Sep;25(3):603–615. doi: 10.1016/0092-8674(81)90168-9. [DOI] [PubMed] [Google Scholar]
  17. Gospodarowicz D., Ill C. Extracellular matrix and control of proliferation of vascular endothelial cells. J Clin Invest. 1980 Jun;65(6):1351–1364. doi: 10.1172/JCI109799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hansson G. K., Björnheden T., Bylock A., Bondjers G. Fc-dependent binding of monocytes to areas with endothelial injury in the rabbit aorta. Exp Mol Pathol. 1981 Jun;34(3):264–280. doi: 10.1016/0014-4800(81)90044-7. [DOI] [PubMed] [Google Scholar]
  19. Hoover R. L., Briggs R. T., Karnovsky M. J. The adhesive interaction between polymorphonuclear leukocytes and endothelial cells in vitro. Cell. 1978 Jun;14(2):423–428. doi: 10.1016/0092-8674(78)90127-7. [DOI] [PubMed] [Google Scholar]
  20. Jauchem J. R., Lopez M., Sprague E. A., Schwartz C. J. Mononuclear cell chemoattractant activity from cultured arterial smooth muscle cells. Exp Mol Pathol. 1982 Oct;37(2):166–174. doi: 10.1016/0014-4800(82)90033-8. [DOI] [PubMed] [Google Scholar]
  21. Joris I., Zand T., Nunnari J. J., Krolikowski F. J., Majno G. Studies on the pathogenesis of atherosclerosis. I. Adhesion and emigration of mononuclear cells in the aorta of hypercholesterolemic rats. Am J Pathol. 1983 Dec;113(3):341–358. [PMC free article] [PubMed] [Google Scholar]
  22. Ladda R. L., Bullock L. P., Gianopoulos T., McCormick L. Radioreceptor assay for epidermal growth factor. Anal Biochem. 1979 Mar;93(2):286–294. doi: 10.1016/s0003-2697(79)80153-0. [DOI] [PubMed] [Google Scholar]
  23. Lewis J. C., Taylor R. G., Jones N. D., St Clair R. W., Cornhill J. F. Endothelial surface characteristics in pigeon coronary artery atherosclerosis. I. Cellular alterations during the initial stages of dietary cholesterol challenge. Lab Invest. 1982 Feb;46(2):123–138. [PubMed] [Google Scholar]
  24. Martin B. M., Gimbrone M. A., Jr, Majeau G. R., Unanue E. R., Cotran R. S. Stimulation of human monocyte/macrophage-derived growth factor (MDGF) production by plasma fibronectin. Am J Pathol. 1983 Jun;111(3):367–373. [PMC free article] [PubMed] [Google Scholar]
  25. Pearson J. D., Carleton J. S., Beesley J. E., Hutchings A., Gordon J. L. Granulocyte adhesion to endothelium in culture. J Cell Sci. 1979 Aug;38:225–235. doi: 10.1242/jcs.38.1.225. [DOI] [PubMed] [Google Scholar]
  26. Ross R. George Lyman Duff Memorial Lecture. Atherosclerosis: a problem of the biology of arterial wall cells and their interactions with blood components. Arteriosclerosis. 1981 Sep-Oct;1(5):293–311. doi: 10.1161/01.atv.1.5.293. [DOI] [PubMed] [Google Scholar]
  27. Ross R. The smooth muscle cell. II. Growth of smooth muscle in culture and formation of elastic fibers. J Cell Biol. 1971 Jul;50(1):172–186. doi: 10.1083/jcb.50.1.172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. STILL W. J., O'NEAL R. M. Electron microscopic study of experimental atherosclerosis in the rat. Am J Pathol. 1962 Jan;40:21–35. [PMC free article] [PubMed] [Google Scholar]
  29. Savion N., Vlodavsky I., Fuks Z. Interaction of T lymphocytes and macrophages with cultured vascular endothelial cells: attachment, invasion, and subsequent degradation of the subendothelial extracellular matrix. J Cell Physiol. 1984 Feb;118(2):169–178. doi: 10.1002/jcp.1041180209. [DOI] [PubMed] [Google Scholar]
  30. Schwartz S. M., Gajdusek C. M., Selden S. C., 3rd Vascular wall growth control: the role of the endothelium. Arteriosclerosis. 1981 Mar-Apr;1(2):107–126. doi: 10.1161/01.atv.1.2.107. [DOI] [PubMed] [Google Scholar]
  31. Sedgwick A. D., Mackay A. R., Bates M. B., Willoughby D. A. The role of plasma factors in the adherence of leucocytes to vascular endothelial cells. J Pathol. 1983 May;140(1):9–16. doi: 10.1002/path.1711400103. [DOI] [PubMed] [Google Scholar]
  32. Stevenson H. C., Miller P., Akiyama Y., Favilla T., Beman J. A., Herberman R., Stull H., Thurman G., Maluish A., Oldham R. A system for obtaining large numbers of cryopreserved human monocytes purified by leukapheresis and counter-current centrifugation elutriation (CCE). J Immunol Methods. 1983 Sep 16;62(3):353–363. doi: 10.1016/0022-1759(83)90180-1. [DOI] [PubMed] [Google Scholar]
  33. Stossel T. P., Pollard T. D., Mason R. J., Vaughan M. Isolation and properties of phagocytic vesicles from polymorphonuclear leukocytes. J Clin Invest. 1971 Aug;50(8):1745–1747. doi: 10.1172/JCI106664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sundström C., Nilsson K. Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer. 1976 May 15;17(5):565–577. doi: 10.1002/ijc.2910170504. [DOI] [PubMed] [Google Scholar]
  35. Walker L. N., Bowyer D. E. Endothelial healing in the rabbit aorta and the effect of risk factors for atherosclerosis. Hypercholesterolemia. Arteriosclerosis. 1984 Sep-Oct;4(5):479–488. doi: 10.1161/01.atv.4.5.479. [DOI] [PubMed] [Google Scholar]
  36. Zweiman B., Moskovitz A. R., Lisak R. P. Quantitative assessment of the adherence of normal human blood mononuclear cells to vascular endothelial cell monolayers. Cell Immunol. 1982 Mar 15;68(1):165–172. doi: 10.1016/0008-8749(82)90099-5. [DOI] [PubMed] [Google Scholar]
  37. de Bono D. P., Green C. The adhesion of different cell types to cultured vascular endothelium: effects of culture density and age. Br J Exp Pathol. 1984 Feb;65(1):145–154. [PMC free article] [PubMed] [Google Scholar]

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