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. 1998 Jun 1;101(11):2427–2437. doi: 10.1172/JCI407

In vivo expression of neutrophil inhibitory factor via gene transfer prevents lipopolysaccharide-induced lung neutrophil infiltration and injury by a beta2 integrin-dependent mechanism.

M Y Zhou 1, S K Lo 1, M Bergenfeldt 1, C Tiruppathi 1, A Jaffe 1, N Xu 1, A B Malik 1
PMCID: PMC508832  PMID: 9616214

Abstract

The binding of beta2 (CD18) integrins on PMN cell membrane to intercellular adhesion molecule (ICAM) counter-receptors on the surface of vascular endothelial cells mediates PMN adhesion to endothelial cells. Neutrophil inhibitory factor (NIF), a 41-kD glycoprotein isolated from the canine hookworm (Ancylostoma caninum), is a beta2 integrin antagonist that inhibits PMN adhesion to endothelial cells. We transferred the NIF gene into CD1 mouse lungs by intravenous injection of cationic liposomes to study the effects of in vivo NIF expression on LPS-induced lung PMN sequestration and the development of lung injury. RT-PCR and Northern blot analysis indicated the lung-selective expression of the NIF transgene, and immunocytochemistry showed prominent NIF expression in pulmonary microvessel endothelial cells. NIF staining was also observed in intraluminal leukocytes present in pulmonary microvessels. This may be the result of NIF binding to leukocytes after its secretion from the transduced lung cells, since there was no evidence of NIF gene expression in circulating leukocytes. Pulmonary vascular NIF expression abrogated the lung tissue PMN uptake and airspace migration of PMN and prevented lung vascular injury (as measured by the lung tissue uptake of [125I]labeled albumin) after the intraperitoneal LPS challenge (200 microg/mouse). Expression of a control protein, chloramphenicol acetyltransferase (CAT), by the same strategy, had no effect on these responses. In vitro studies showed that NIF prevented mouse PMN adhesion consistent with the inhibition of lung uptake after LPS challenge in NIF transgene-expressing mice. We conclude that pulmonary vascular expression of NIF, a specific beta2 integrin- binding protein, is a potentially useful gene transfer strategy in modulating the infiltration of PMN across the alveolar-capillary epithelial barrier and in preventing lung vascular endothelial injury.

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Selected References

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  1. Anderson D. C., Springer T. A. Leukocyte adhesion deficiency: an inherited defect in the Mac-1, LFA-1, and p150,95 glycoproteins. Annu Rev Med. 1987;38:175–194. doi: 10.1146/annurev.me.38.020187.001135. [DOI] [PubMed] [Google Scholar]
  2. Arnaout M. A., Gupta S. K., Pierce M. W., Tenen D. G. Amino acid sequence of the alpha subunit of human leukocyte adhesion receptor Mo1 (complement receptor type 3). J Cell Biol. 1988 Jun;106(6):2153–2158. doi: 10.1083/jcb.106.6.2153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arnaout M. A. Structure and function of the leukocyte adhesion molecules CD11/CD18. Blood. 1990 Mar 1;75(5):1037–1050. [PubMed] [Google Scholar]
  4. Barnard J. W., Biro M. G., Lo S. K., Ohno S., Carozza M. A., Moyle M., Soule H. R., Malik A. B. Neutrophil inhibitory factor prevents neutrophil-dependent lung injury. J Immunol. 1995 Nov 15;155(10):4876–4881. [PubMed] [Google Scholar]
  5. Brigham K. L., Meyrick B. Endotoxin and lung injury. Am Rev Respir Dis. 1986 May;133(5):913–927. [PubMed] [Google Scholar]
  6. Curiel D. T., Pilewski J. M., Albelda S. M. Gene therapy approaches for inherited and acquired lung diseases. Am J Respir Cell Mol Biol. 1996 Jan;14(1):1–18. doi: 10.1165/ajrcmb.14.1.8534480. [DOI] [PubMed] [Google Scholar]
  7. Diamond M. S., Garcia-Aguilar J., Bickford J. K., Corbi A. L., Springer T. A. The I domain is a major recognition site on the leukocyte integrin Mac-1 (CD11b/CD18) for four distinct adhesion ligands. J Cell Biol. 1993 Feb;120(4):1031–1043. doi: 10.1083/jcb.120.4.1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eichacker P. Q., Hoffman W. D., Farese A., Danner R. L., Suffredini A. F., Waisman Y., Banks S. M., Mouginis T., Wilson L., Rothlein R. Leukocyte CD18 monoclonal antibody worsens endotoxemia and cardiovascular injury in canines with septic shock. J Appl Physiol (1985) 1993 Apr;74(4):1885–1892. doi: 10.1152/jappl.1993.74.4.1885. [DOI] [PubMed] [Google Scholar]
  9. Furie M. B., Tancinco M. C., Smith C. W. Monoclonal antibodies to leukocyte integrins CD11a/CD18 and CD11b/CD18 or intercellular adhesion molecule-1 inhibit chemoattractant-stimulated neutrophil transendothelial migration in vitro. Blood. 1991 Oct 15;78(8):2089–2097. [PubMed] [Google Scholar]
  10. Gao X., Huang L. Cationic liposome-mediated gene transfer. Gene Ther. 1995 Dec;2(10):710–722. [PubMed] [Google Scholar]
  11. Glauser M. P., Zanetti G., Baumgartner J. D., Cohen J. Septic shock: pathogenesis. Lancet. 1991 Sep 21;338(8769):732–736. doi: 10.1016/0140-6736(91)91452-z. [DOI] [PubMed] [Google Scholar]
  12. Graf J. M., Smith C. W., Mariscalco M. M. Contribution of LFA-1 and Mac-1 to CD18-dependent neutrophil emigration in a neonatal rabbit model. J Appl Physiol (1985) 1996 Jun;80(6):1984–1992. doi: 10.1152/jappl.1996.80.6.1984. [DOI] [PubMed] [Google Scholar]
  13. Green T. P., Johnson D. E., Marchessault R. P., Gatto C. W. Transvascular flux and tissue accrual of Evans blue: effects of endotoxin and histamine. J Lab Clin Med. 1988 Feb;111(2):173–183. [PubMed] [Google Scholar]
  14. Harlan J. M. Neutrophil-mediated vascular injury. Acta Med Scand Suppl. 1987;715:123–129. doi: 10.1111/j.0954-6820.1987.tb09912.x. [DOI] [PubMed] [Google Scholar]
  15. Jin F. Y., Nathan C., Radzioch D., Ding A. Secretory leukocyte protease inhibitor: a macrophage product induced by and antagonistic to bacterial lipopolysaccharide. Cell. 1997 Feb 7;88(3):417–426. doi: 10.1016/s0092-8674(00)81880-2. [DOI] [PubMed] [Google Scholar]
  16. Krawisz J. E., Sharon P., Stenson W. F. Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models. Gastroenterology. 1984 Dec;87(6):1344–1350. [PubMed] [Google Scholar]
  17. Lee J. O., Rieu P., Arnaout M. A., Liddington R. Crystal structure of the A domain from the alpha subunit of integrin CR3 (CD11b/CD18). Cell. 1995 Feb 24;80(4):631–638. doi: 10.1016/0092-8674(95)90517-0. [DOI] [PubMed] [Google Scholar]
  18. Liu Y., Liggitt D., Zhong W., Tu G., Gaensler K., Debs R. Cationic liposome-mediated intravenous gene delivery. J Biol Chem. 1995 Oct 20;270(42):24864–24870. doi: 10.1074/jbc.270.42.24864. [DOI] [PubMed] [Google Scholar]
  19. Lo S. K., Van Seventer G. A., Levin S. M., Wright S. D. Two leukocyte receptors (CD11a/CD18 and CD11b/CD18) mediate transient adhesion to endothelium by binding to different ligands. J Immunol. 1989 Nov 15;143(10):3325–3329. [PubMed] [Google Scholar]
  20. Lu H., Smith C. W., Perrard J., Bullard D., Tang L., Shappell S. B., Entman M. L., Beaudet A. L., Ballantyne C. M. LFA-1 is sufficient in mediating neutrophil emigration in Mac-1-deficient mice. J Clin Invest. 1997 Mar 15;99(6):1340–1350. doi: 10.1172/JCI119293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McCandless B. K., Kaufman R. P., Jr, Cooper J. A., Neumann P. H., Malik A. B. Mediation of lung neutrophil uptake after endotoxin by CD18-integrin-dependent and -independent mechanisms. Am J Physiol. 1994 Apr;266(4 Pt 2):H1451–H1456. doi: 10.1152/ajpheart.1994.266.4.H1451. [DOI] [PubMed] [Google Scholar]
  22. McLean J. W., Fox E. A., Baluk P., Bolton P. B., Haskell A., Pearlman R., Thurston G., Umemoto E. Y., McDonald D. M. Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice. Am J Physiol. 1997 Jul;273(1 Pt 2):H387–H404. doi: 10.1152/ajpheart.1997.273.1.H387. [DOI] [PubMed] [Google Scholar]
  23. Miotla J. M., Williams T. J., Hellewell P. G., Jeffery P. K. A role for the beta2 integrin CD11b in mediating experimental lung injury in mice. Am J Respir Cell Mol Biol. 1996 Apr;14(4):363–373. doi: 10.1165/ajrcmb.14.4.8600941. [DOI] [PubMed] [Google Scholar]
  24. Morisaki T., Goya T., Toh H., Nishihara K., Torisu M. The anti Mac-1 monoclonal antibody inhibits neutrophil sequestration in lung and liver in a septic murine model. Clin Immunol Immunopathol. 1991 Dec;61(3):365–375. doi: 10.1016/s0090-1229(05)80008-x. [DOI] [PubMed] [Google Scholar]
  25. Moyle M., Foster D. L., McGrath D. E., Brown S. M., Laroche Y., De Meutter J., Stanssens P., Bogowitz C. A., Fried V. A., Ely J. A. A hookworm glycoprotein that inhibits neutrophil function is a ligand of the integrin CD11b/CD18. J Biol Chem. 1994 Apr 1;269(13):10008–10015. [PubMed] [Google Scholar]
  26. Muchowski P. J., Zhang L., Chang E. R., Soule H. R., Plow E. F., Moyle M. Functional interaction between the integrin antagonist neutrophil inhibitory factor and the I domain of CD11b/CD18. J Biol Chem. 1994 Oct 21;269(42):26419–26423. [PubMed] [Google Scholar]
  27. Muller D. W., Gordon D., San H., Yang Z., Pompili V. J., Nabel G. J., Nabel E. G. Catheter-mediated pulmonary vascular gene transfer and expression. Circ Res. 1994 Dec;75(6):1039–1049. doi: 10.1161/01.res.75.6.1039. [DOI] [PubMed] [Google Scholar]
  28. Rieu P., Ueda T., Haruta I., Sharma C. P., Arnaout M. A. The A-domain of beta 2 integrin CR3 (CD11b/CD18) is a receptor for the hookworm-derived neutrophil adhesion inhibitor NIF. J Cell Biol. 1994 Dec;127(6 Pt 2):2081–2091. doi: 10.1083/jcb.127.6.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rothlein R., Dustin M. L., Marlin S. D., Springer T. A. A human intercellular adhesion molecule (ICAM-1) distinct from LFA-1. J Immunol. 1986 Aug 15;137(4):1270–1274. [PubMed] [Google Scholar]
  30. Saltini C., Hance A. J., Ferrans V. J., Basset F., Bitterman P. B., Crystal R. G. Accurate quantification of cells recovered by bronchoalveolar lavage. Am Rev Respir Dis. 1984 Oct;130(4):650–658. doi: 10.1164/arrd.1984.130.4.650. [DOI] [PubMed] [Google Scholar]
  31. Springer T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994 Jan 28;76(2):301–314. doi: 10.1016/0092-8674(94)90337-9. [DOI] [PubMed] [Google Scholar]
  32. Standiford T. J., Kunkel S. L., Lukacs N. W., Greenberger M. J., Danforth J. M., Kunkel R. G., Strieter R. M. Macrophage inflammatory protein-1 alpha mediates lung leukocyte recruitment, lung capillary leak, and early mortality in murine endotoxemia. J Immunol. 1995 Aug 1;155(3):1515–1524. [PubMed] [Google Scholar]
  33. Tang W. W., Yi E. S., Remick D. G., Wittwer A., Yin S., Qi M., Ulich T. R. Intratracheal injection of endotoxin and cytokines. IX. Contribution of CD11a/ICAM-1 to neutrophil emigration. Am J Physiol. 1995 Nov;269(5 Pt 1):L653–L659. doi: 10.1152/ajplung.1995.269.5.L653. [DOI] [PubMed] [Google Scholar]
  34. Tate R. M., Repine J. E. Neutrophils and the adult respiratory distress syndrome. Am Rev Respir Dis. 1983 Sep;128(3):552–559. doi: 10.1164/arrd.1983.128.3.552. [DOI] [PubMed] [Google Scholar]
  35. Vedder N. B., Winn R. K., Rice C. L., Harlan J. M. Neutrophil-mediated vascular injury in shock and multiple organ failure. Prog Clin Biol Res. 1989;299:181–191. [PubMed] [Google Scholar]
  36. Xie J., Li R., Kotovuori P., Vermot-Desroches C., Wijdenes J., Arnaout M. A., Nortamo P., Gahmberg C. G. Intercellular adhesion molecule-2 (CD102) binds to the leukocyte integrin CD11b/CD18 through the A domain. J Immunol. 1995 Oct 1;155(7):3619–3628. [PubMed] [Google Scholar]
  37. Zhou L., Lee D. H., Plescia J., Lau C. Y., Altieri D. C. Differential ligand binding specificities of recombinant CD11b/CD18 integrin I-domain. J Biol Chem. 1994 Jun 24;269(25):17075–17079. [PubMed] [Google Scholar]

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