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. 1997 May;150(5):1673–1685.

Vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) in normal and atherosclerotic human arteries.

T Couffinhal 1, M Kearney 1, B Witzenbichler 1, D Chen 1, T Murohara 1, D W Losordo 1, J Symes 1, J M Isner 1
PMCID: PMC1858200  PMID: 9137092

Abstract

Vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) is an endothelial-cell-specific mitogen; as such, its role in angiogenesis has been studied extensively. VEGF/VPF may also serve as a local, endogenous regulator of large-vessel endothelial cell integrity. Surprisingly, however, VEGF/VPF expression in normal and/or atherosclerotic vessels has not been previously characterized. Accordingly, we studied normal human arteries and veins as well as atherosclerotic and restenotic human coronary arteries for evidence of VEGF/VPF expression. VEGF/VPF was detected immunohistochemically in sections of normal human aorta, mammary artery, and saphenous vein. Moreover, VEGF/ VPF expression was identified in 32 (97%) of 33 pathological coronary arterial specimens; the extent of VEGF/VPF staining was graded as moderate to strong in 21 of the 32 (66%) positive specimens. VEGF/VPF double immunostaining and in situ hybridization demonstrated that smooth muscle cells constitute the principal cellular source of VEGF/VPF. VEGF/VPF immunostaining among primary atherosclerotic lesions localized predominantly to the extracellular matrix. In restenotic specimens, VEGF/VPF immunostaining was more prominently cellular, particularly among proliferating smooth muscle cells. Although VEGF/VPF expression was observed in areas of macrophage infiltration, double immunostaining failed to localize VEGF/VPF to macrophages in these foci; instead, double immunostaining clearly identified CD45RO-positive cells as responsible for VEGF/VPF expression in such areas. No correlation could be demonstrated between VEGF/VPF immunostaining and extent of vasa vasorum. These findings thus establish that postnatal VEGF/VPF expression is a feature of normal human arteries and veins and is often extensively expressed in arteries narrowed by atherosclerotic plaque. VEGF/VPF expression in the wall and/or plaque of medium to large vessels suggests a role for VEGF/VPF other than promoting angiogenesis. This role may involve maintenance and repair of luminal endothelium.

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Selected References

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

  1. Akbar A. N., Terry L., Timms A., Beverley P. C., Janossy G. Loss of CD45R and gain of UCHL1 reactivity is a feature of primed T cells. J Immunol. 1988 Apr 1;140(7):2171–2178. [PubMed] [Google Scholar]
  2. Alon T., Hemo I., Itin A., Pe'er J., Stone J., Keshet E. Vascular endothelial growth factor acts as a survival factor for newly formed retinal vessels and has implications for retinopathy of prematurity. Nat Med. 1995 Oct;1(10):1024–1028. doi: 10.1038/nm1095-1024. [DOI] [PubMed] [Google Scholar]
  3. Asahara T., Bauters C., Pastore C., Kearney M., Rossow S., Bunting S., Ferrara N., Symes J. F., Isner J. M. Local delivery of vascular endothelial growth factor accelerates reendothelialization and attenuates intimal hyperplasia in balloon-injured rat carotid artery. Circulation. 1995 Jun 1;91(11):2793–2801. doi: 10.1161/01.cir.91.11.2793. [DOI] [PubMed] [Google Scholar]
  4. Asahara T., Chen D., Tsurumi Y., Kearney M., Rossow S., Passeri J., Symes J. F., Isner J. M. Accelerated restitution of endothelial integrity and endothelium-dependent function after phVEGF165 gene transfer. Circulation. 1996 Dec 15;94(12):3291–3302. doi: 10.1161/01.cir.94.12.3291. [DOI] [PubMed] [Google Scholar]
  5. Berse B., Brown L. F., Van de Water L., Dvorak H. F., Senger D. R. Vascular permeability factor (vascular endothelial growth factor) gene is expressed differentially in normal tissues, macrophages, and tumors. Mol Biol Cell. 1992 Feb;3(2):211–220. doi: 10.1091/mbc.3.2.211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Breier G., Albrecht U., Sterrer S., Risau W. Expression of vascular endothelial growth factor during embryonic angiogenesis and endothelial cell differentiation. Development. 1992 Feb;114(2):521–532. doi: 10.1242/dev.114.2.521. [DOI] [PubMed] [Google Scholar]
  7. Brogi E., Schatteman G., Wu T., Kim E. A., Varticovski L., Keyt B., Isner J. M. Hypoxia-induced paracrine regulation of vascular endothelial growth factor receptor expression. J Clin Invest. 1996 Jan 15;97(2):469–476. doi: 10.1172/JCI118437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brogi E., Wu T., Namiki A., Isner J. M. Indirect angiogenic cytokines upregulate VEGF and bFGF gene expression in vascular smooth muscle cells, whereas hypoxia upregulates VEGF expression only. Circulation. 1994 Aug;90(2):649–652. doi: 10.1161/01.cir.90.2.649. [DOI] [PubMed] [Google Scholar]
  9. Brown L. F., Berse B., Jackman R. W., Tognazzi K., Guidi A. J., Dvorak H. F., Senger D. R., Connolly J. L., Schnitt S. J. Expression of vascular permeability factor (vascular endothelial growth factor) and its receptors in breast cancer. Hum Pathol. 1995 Jan;26(1):86–91. doi: 10.1016/0046-8177(95)90119-1. [DOI] [PubMed] [Google Scholar]
  10. Carmeliet P., Ferreira V., Breier G., Pollefeyt S., Kieckens L., Gertsenstein M., Fahrig M., Vandenhoeck A., Harpal K., Eberhardt C. Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature. 1996 Apr 4;380(6573):435–439. doi: 10.1038/380435a0. [DOI] [PubMed] [Google Scholar]
  11. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  12. Detmar M., Brown L. F., Claffey K. P., Yeo K. T., Kocher O., Jackman R. W., Berse B., Dvorak H. F. Overexpression of vascular permeability factor/vascular endothelial growth factor and its receptors in psoriasis. J Exp Med. 1994 Sep 1;180(3):1141–1146. doi: 10.1084/jem.180.3.1141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Drake C. J., Little C. D. Exogenous vascular endothelial growth factor induces malformed and hyperfused vessels during embryonic neovascularization. Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7657–7661. doi: 10.1073/pnas.92.17.7657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dvorak H. F., Brown L. F., Detmar M., Dvorak A. M. Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am J Pathol. 1995 May;146(5):1029–1039. [PMC free article] [PubMed] [Google Scholar]
  15. Dvorak H. F., Sioussat T. M., Brown L. F., Berse B., Nagy J. A., Sotrel A., Manseau E. J., Van de Water L., Senger D. R. Distribution of vascular permeability factor (vascular endothelial growth factor) in tumors: concentration in tumor blood vessels. J Exp Med. 1991 Nov 1;174(5):1275–1278. doi: 10.1084/jem.174.5.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ferguson J. J. American College of Cardiology 45th Annual Scientific Session, Orlando, Florida, March 24 to 27, 1996. Circulation. 1996 Jul 1;94(1):1–5. doi: 10.1161/01.cir.94.1.1. [DOI] [PubMed] [Google Scholar]
  17. Ferrara N., Carver-Moore K., Chen H., Dowd M., Lu L., O'Shea K. S., Powell-Braxton L., Hillan K. J., Moore M. W. Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene. Nature. 1996 Apr 4;380(6573):439–442. doi: 10.1038/380439a0. [DOI] [PubMed] [Google Scholar]
  18. Flamme I., von Reutern M., Drexler H. C., Syed-Ali S., Risau W. Overexpression of vascular endothelial growth factor in the avian embryo induces hypervascularization and increased vascular permeability without alterations of embryonic pattern formation. Dev Biol. 1995 Oct;171(2):399–414. doi: 10.1006/dbio.1995.1291. [DOI] [PubMed] [Google Scholar]
  19. Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med. 1995 Jan;1(1):27–31. doi: 10.1038/nm0195-27. [DOI] [PubMed] [Google Scholar]
  20. Freeman M. R., Schneck F. X., Gagnon M. L., Corless C., Soker S., Niknejad K., Peoples G. E., Klagsbrun M. Peripheral blood T lymphocytes and lymphocytes infiltrating human cancers express vascular endothelial growth factor: a potential role for T cells in angiogenesis. Cancer Res. 1995 Sep 15;55(18):4140–4145. [PubMed] [Google Scholar]
  21. Fuster V., Badimon L., Badimon J. J., Chesebro J. H. The pathogenesis of coronary artery disease and the acute coronary syndromes (2). N Engl J Med. 1992 Jan 30;326(5):310–318. doi: 10.1056/NEJM199201303260506. [DOI] [PubMed] [Google Scholar]
  22. Hashimoto M., Ohsawa M., Ohnishi A., Naka N., Hirota S., Kitamura Y., Aozasa K. Expression of vascular endothelial growth factor and its receptor mRNA in angiosarcoma. Lab Invest. 1995 Dec;73(6):859–863. [PubMed] [Google Scholar]
  23. Hatva E., Böhling T., Jäskeläinen J., Persico M. G., Haltia M., Alitalo K. Vascular growth factors and receptors in capillary hemangioblastomas and hemangiopericytomas. Am J Pathol. 1996 Mar;148(3):763–775. [PMC free article] [PubMed] [Google Scholar]
  24. Iijima K., Yoshikawa N., Connolly D. T., Nakamura H. Human mesangial cells and peripheral blood mononuclear cells produce vascular permeability factor. Kidney Int. 1993 Nov;44(5):959–966. doi: 10.1038/ki.1993.337. [DOI] [PubMed] [Google Scholar]
  25. Ikeda E., Achen M. G., Breier G., Risau W. Hypoxia-induced transcriptional activation and increased mRNA stability of vascular endothelial growth factor in C6 glioma cells. J Biol Chem. 1995 Aug 25;270(34):19761–19766. doi: 10.1074/jbc.270.34.19761. [DOI] [PubMed] [Google Scholar]
  26. Isner J. M., Kearney M., Bortman S., Passeri J. Apoptosis in human atherosclerosis and restenosis. Circulation. 1995 Jun 1;91(11):2703–2711. doi: 10.1161/01.cir.91.11.2703. [DOI] [PubMed] [Google Scholar]
  27. Isner J. M., Walsh K., Rosenfield K., Schainfeld R., Asahara T., Hogan K., Pieczek A. Arterial gene therapy for restenosis. Hum Gene Ther. 1996 May 20;7(8):989–1011. doi: 10.1089/hum.1996.7.8-989. [DOI] [PubMed] [Google Scholar]
  28. Ito A., Nomura S., Hirota S., Suda J., Suda T., Kitamura Y. Enhanced expression of CD34 messenger RNA by developing endothelial cells of mice. Lab Invest. 1995 May;72(5):532–538. [PubMed] [Google Scholar]
  29. Jakeman L. B., Winer J., Bennett G. L., Altar C. A., Ferrara N. Binding sites for vascular endothelial growth factor are localized on endothelial cells in adult rat tissues. J Clin Invest. 1992 Jan;89(1):244–253. doi: 10.1172/JCI115568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Katoh O., Tauchi H., Kawaishi K., Kimura A., Satow Y. Expression of the vascular endothelial growth factor (VEGF) receptor gene, KDR, in hematopoietic cells and inhibitory effect of VEGF on apoptotic cell death caused by ionizing radiation. Cancer Res. 1995 Dec 1;55(23):5687–5692. [PubMed] [Google Scholar]
  31. Koch A. E., Harlow L. A., Haines G. K., Amento E. P., Unemori E. N., Wong W. L., Pope R. M., Ferrara N. Vascular endothelial growth factor. A cytokine modulating endothelial function in rheumatoid arthritis. J Immunol. 1994 Apr 15;152(8):4149–4156. [PubMed] [Google Scholar]
  32. Ku D. D., Zaleski J. K., Liu S., Brock T. A. Vascular endothelial growth factor induces EDRF-dependent relaxation in coronary arteries. Am J Physiol. 1993 Aug;265(2 Pt 2):H586–H592. doi: 10.1152/ajpheart.1993.265.2.H586. [DOI] [PubMed] [Google Scholar]
  33. Lüscher T. F., Tanner F. C., Tschudi M. R., Noll G. Endothelial dysfunction in coronary artery disease. Annu Rev Med. 1993;44:395–418. doi: 10.1146/annurev.me.44.020193.002143. [DOI] [PubMed] [Google Scholar]
  34. Millauer B., Wizigmann-Voos S., Schnürch H., Martinez R., Møller N. P., Risau W., Ullrich A. High affinity VEGF binding and developmental expression suggest Flk-1 as a major regulator of vasculogenesis and angiogenesis. Cell. 1993 Mar 26;72(6):835–846. doi: 10.1016/0092-8674(93)90573-9. [DOI] [PubMed] [Google Scholar]
  35. Monacci W. T., Merrill M. J., Oldfield E. H. Expression of vascular permeability factor/vascular endothelial growth factor in normal rat tissues. Am J Physiol. 1993 Apr;264(4 Pt 1):C995–1002. doi: 10.1152/ajpcell.1993.264.4.C995. [DOI] [PubMed] [Google Scholar]
  36. Namiki A., Brogi E., Kearney M., Kim E. A., Wu T., Couffinhal T., Varticovski L., Isner J. M. Hypoxia induces vascular endothelial growth factor in cultured human endothelial cells. J Biol Chem. 1995 Dec 29;270(52):31189–31195. doi: 10.1074/jbc.270.52.31189. [DOI] [PubMed] [Google Scholar]
  37. Ohsawa M., Naka N., Tomita Y., Kawamori D., Kanno H., Aozasa K. Use of immunohistochemical procedures in diagnosing angiosarcoma. Evaluation of 98 cases. Cancer. 1995 Jun 15;75(12):2867–2874. doi: 10.1002/1097-0142(19950615)75:12<2867::aid-cncr2820751212>3.0.co;2-8. [DOI] [PubMed] [Google Scholar]
  38. Park J. E., Keller G. A., Ferrara N. The vascular endothelial growth factor (VEGF) isoforms: differential deposition into the subepithelial extracellular matrix and bioactivity of extracellular matrix-bound VEGF. Mol Biol Cell. 1993 Dec;4(12):1317–1326. doi: 10.1091/mbc.4.12.1317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Peters K. G., De Vries C., Williams L. T. Vascular endothelial growth factor receptor expression during embryogenesis and tissue repair suggests a role in endothelial differentiation and blood vessel growth. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8915–8919. doi: 10.1073/pnas.90.19.8915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Plate K. H., Breier G., Millauer B., Ullrich A., Risau W. Up-regulation of vascular endothelial growth factor and its cognate receptors in a rat glioma model of tumor angiogenesis. Cancer Res. 1993 Dec 1;53(23):5822–5827. [PubMed] [Google Scholar]
  41. Plate K. H., Breier G., Weich H. A., Mennel H. D., Risau W. Vascular endothelial growth factor and glioma angiogenesis: coordinate induction of VEGF receptors, distribution of VEGF protein and possible in vivo regulatory mechanisms. Int J Cancer. 1994 Nov 15;59(4):520–529. doi: 10.1002/ijc.2910590415. [DOI] [PubMed] [Google Scholar]
  42. Plate K. H., Breier G., Weich H. A., Risau W. Vascular endothelial growth factor is a potential tumour angiogenesis factor in human gliomas in vivo. Nature. 1992 Oct 29;359(6398):845–848. doi: 10.1038/359845a0. [DOI] [PubMed] [Google Scholar]
  43. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  44. Senger D. R., Galli S. J., Dvorak A. M., Perruzzi C. A., Harvey V. S., Dvorak H. F. Tumor cells secrete a vascular permeability factor that promotes accumulation of ascites fluid. Science. 1983 Feb 25;219(4587):983–985. doi: 10.1126/science.6823562. [DOI] [PubMed] [Google Scholar]
  45. Senger D. R., Ledbetter S. R., Claffey K. P., Papadopoulos-Sergiou A., Peruzzi C. A., Detmar M. Stimulation of endothelial cell migration by vascular permeability factor/vascular endothelial growth factor through cooperative mechanisms involving the alphavbeta3 integrin, osteopontin, and thrombin. Am J Pathol. 1996 Jul;149(1):293–305. [PMC free article] [PubMed] [Google Scholar]
  46. Shifren J. L., Doldi N., Ferrara N., Mesiano S., Jaffe R. B. In the human fetus, vascular endothelial growth factor is expressed in epithelial cells and myocytes, but not vascular endothelium: implications for mode of action. J Clin Endocrinol Metab. 1994 Jul;79(1):316–322. doi: 10.1210/jcem.79.1.8027247. [DOI] [PubMed] [Google Scholar]
  47. Shweiki D., Itin A., Soffer D., Keshet E. Vascular endothelial growth factor induced by hypoxia may mediate hypoxia-initiated angiogenesis. Nature. 1992 Oct 29;359(6398):843–845. doi: 10.1038/359843a0. [DOI] [PubMed] [Google Scholar]
  48. Smith S. C., Jr AHA president's letter. Circulation. 1995 Jul 1;92(1):1–1. [PubMed] [Google Scholar]
  49. Stavri G. T., Zachary I. C., Baskerville P. A., Martin J. F., Erusalimsky J. D. Basic fibroblast growth factor upregulates the expression of vascular endothelial growth factor in vascular smooth muscle cells. Synergistic interaction with hypoxia. Circulation. 1995 Jul 1;92(1):11–14. doi: 10.1161/01.cir.92.1.11. [DOI] [PubMed] [Google Scholar]
  50. Tischer E., Mitchell R., Hartman T., Silva M., Gospodarowicz D., Fiddes J. C., Abraham J. A. The human gene for vascular endothelial growth factor. Multiple protein forms are encoded through alternative exon splicing. J Biol Chem. 1991 Jun 25;266(18):11947–11954. [PubMed] [Google Scholar]
  51. Van Belle E., Tio F. O., Couffinhal T., Maillard L., Passeri J., Isner J. M. Stent endothelialization. Time course, impact of local catheter delivery, feasibility of recombinant protein administration, and response to cytokine expedition. Circulation. 1997 Jan 21;95(2):438–448. doi: 10.1161/01.cir.95.2.438. [DOI] [PubMed] [Google Scholar]
  52. Wizigmann-Voos S., Breier G., Risau W., Plate K. H. Up-regulation of vascular endothelial growth factor and its receptors in von Hippel-Lindau disease-associated and sporadic hemangioblastomas. Cancer Res. 1995 Mar 15;55(6):1358–1364. [PubMed] [Google Scholar]
  53. van der Zee R., Murohara T., Luo Z., Zollmann F., Passeri J., Lekutat C., Isner J. M. Vascular endothelial growth factor/vascular permeability factor augments nitric oxide release from quiescent rabbit and human vascular endothelium. Circulation. 1997 Feb 18;95(4):1030–1037. doi: 10.1161/01.cir.95.4.1030. [DOI] [PubMed] [Google Scholar]

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