Abstract
Integrin subunits αv and β3 form a dimer, αvβ3, which is expressed on normal neutrophils and endothelium. We investigated the expression of integrin subunits αv and β3 in acute lung inflammation in Sprague-Dawley rats (n=5 each) following intratracheal challenge with Escherichia coli or Streptococcus pneumoniae, which induce neutrophil recruitment through different mechanisms. Control rats (n=5) were given endotoxin-free saline. Both bacterial challenges induced similar levels of recruitment of neutrophils in lungs. Western blots showed lower expression of integrin subunits αv and β3 in lungs challenged with E. coli compared to those given S. pneumoniae. Immunohistochemistry and immunogold electron microscopy localized both integrin subunits in neutrophils and endothelium in the control and treated rat lungs. Quantitative immunohistochemistry showed that E. coli-challenged rat lungs contained a lower percentage of neutrophils expressing integrin subunits αv and β3 compared to those challenged with S. pneumoniae (P<0.05). We conclude that E. coli infection decreased the percentage of neutrophils expressing integrin subunits αv and β3 compared to S. pneumoniae infection. These data lay the foundation for further characterization of these integrin subunits in neutrophil migration specifically in S. pneumoniae infection that utilizes molecules other than β2 integrins for neutrophil recruitment.
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References
Berton G, Lowell CA (1999) Integrin signalling in neutrophils and macrophages. Cell Signal 11:621–635
Bhattacharya S, Fu C, Bhattacharya J, Greenberg S (1995) Soluble ligands of the αvβ3 integrin mediate enhanced tyrosine phosphorylation of multiple proteins in adherent bovine pulmonary artery endothelial cells. J Biol Chem 270:16781–16787
Bhattacharya S, Ying X, Fu C, Patel R, Kuebler WM, Greenberg S, Bhattacharya J (2000) alpha(v)beta(3) integrin induces tyrosine phosphorylation-dependent Ca(2+) influx in pulmonary endothelial cells. Circ Res 86:456–462
Bhattacharya S, Patel R, Sen N, Quadri S, Parthasarathi K, Bhattacharya J (2001) Dual signaling by the αvβ3-integrin activates cytosolic PLA2 in bovine pulmonary artery endothelial cells. Am J Physiol 280:L1049–L1056
Brooks PC, Clark RAF, Cheresh DA (1994) Requirement of vascular integrin αvβ3 for angiogenesis. Science 264:569–571
Bruyninckx WJ, Comerford KM, Lawrence DW, Colgan SP (2001) Phosphoinositide 3-kinase modulation of β3-integrin represents and endogenous “braking” mechanism during neutrophil transmatrix migration. Blood 97:3251–3258
Burns AR, Smith CW, Walker DC (2003) Unique structural features that influence neutrophil emigration into the lung. Physiol Rev 83:309–336
Conforti G, Dominguez-Jimenez C, Zanetti A, Gimbrone MA Jr, Cremona O, Marchisio PC, Dejana E (1992) Human endothelial cells expressing integrin receptors are on the luminal aspect of their membrane. Blood 80:437–446
Damjanovich L, Albelda SM, Mette SA, Buck CA (1992) Distribution of integrin cell adhesion receptors in normal and malignant lung tissue. Am J Respir Cell Mol Biol 6:197–206
Doerschuk CM, Winn RK, Coxson HO, Harlan JM (1990) CD18-dependent and -independent mechanisms of neutrophil emigration in the pulmonary and systemic microcirculation of rabbits. J Immunol 144:2327–2333
Hendey B, Lawson M, Morcantonio EE, Maxfield FR (1996) Intracellular calcium and calcineurin regulate neutrophil motility on vitronectin through a receptor identified by antibodies to integrins αv and β3. Blood 87:2038–2048
Hogg JC, Doerschuk CM (1995) Leukocyte traffic in the lung. Annu Rev Physiol 57:97–114
Horton MA (1997) The alpha v beta 3 integrin “vitronectin receptor”. Int J Biochem Cell Biol 29:721–725
Koukoulis GK, Warren WH, Virtanen I, Gould VE (1997) Immunolocalization of integrins in the normal lung and in pulmonary carcinomas. Hum Pathol 28:1018–1025
Langlois PF, Gawryl MS (1988) Accentuated formation of the terminal C5b-9 complement complex in patient plasma precedes development of the adult respiratory distress syndrome. Am Rev Respir Dis 138:368–375
Lawson MA, Maxfield FR (1995) Ca2+- and calcineurin-dependent recycling of an integrin to the front of migrating neutrophils. Nature 377:75–79
Miller LJ, Bainton DF, Borregaard N, Springer TA (1987) Stimulated mobilization of monocyte Mac-1 and p150,95 adhesion proteins from an intracellular vesicular compartment to the cell surface. J Clin Invest 80:535–544
Mizgerd JP (2002) Molecular mechanisms of neutrophil recruitment elicited by bacteria in the lungs. Semin Immunol 14:123–132
Mizgerd JP, Kubo H, Kutkoski GJ, Bhagwan SD, Scharffetter-Kochanek K, Beaudet AL, Doerschuk CM (1997) Neutrophil emigration in the skin, lungs, and peritoneum: different requirements for CD11/CD18 revealed by CD18-deficient mice. J Exp Med 186:1357–1362
Porter JC, Hogg N (1997) Integrin cross talk: activation of lymphocyte function-associated antigen-1 on human T cells alters alpha 4beta 1- and alpha 5beta 1-mediated function. J Cell Biol 138:1437–1447
Rainger GE, Buckley CD, Simmons DL, Nash GB (1999) Neutrophils sense flow-generated stress and direct their migration through αvβ3-integrin. Am J Physiol 276:H858–H864
Retta SF, Cassara G, D’Amato M, Alessandro R, Pellegrino M, Degani S, De Leo G, Silengo L, Tarone G (2001) Cross talk between beta 1 and alpha V integrins: beta 1 affects beta 3 mRNA stability. Mol Biol Cell 12:3126–3138
Seiffert D (1997) Constitutive and regulated expression of vitronectin. Histol Histopathol 12:787–797
Seiffert D, Smith JW (1997) The cell adhesion domain in plasma vitronectin is cryptic. J Biol Chem 272:13705–13710
Singh B, Fu C, Bhattacharya J (2000) Vascular expression of the αvβ3 integrin in lung and other organs. Am J Physiol 278:L217–L226
Sixt M, Hallmann R, Wendler O, Scharffetter-Kochanek K, Sorokin LM (2001) Cell adhesion and migration properties of β2-integrin negative polymorphonuclear granulocytes on defined extracellular matrix molecules. J Biol Chem 276:18878–18887
Thompson RD, Wakelin MW, Larbi KY, Dewar A, Asimakopoulos G, Horton MA, Nakada MT, Nourshargh S (2000) Divergent effects of platelet-endothelial cell adhesion molecule-1 and beta 3 integrin blockade on leukocyte transmigration in vivo. J Immunol 165:426–434
Tsukada H, Ying X, Fu C, Ishikawa S, McKeown-Longo PJ, Albelda SM, Bhattacharya S, Bray BA, Bhattacharya J (1995) Ligation of endothelial αvβ3 integrin increases capillary hydraulic conductivity of rat lung. Circ Res 77:651–659
van der Flier A, Sonnenberg A (2001) Function and interactions of integrins. Cell Tissue Res 305:285–298
von Andrian UH, Chambers JD, McEvoy LM, Bargatze RF, Arfors K, Butcher EC (1991) Two-step model of leukocyte–endothelial cell interaction in inflammation: distinct roles for LECAM-1 and the leukocyte {beta}2 integrins in vivo. Proc Natl Acad Sci U S A 88:7538–7542
Ware LB, Matthay MA (2000) The acute respiratory distress syndrome. N Engl J Med 342:1334–1349
Weerasinghe D, McHugh KP, Ross FP, Brown EJ, Gisler RH, Imhof BA (1998) A role of the αvβ3 integrin in the transmigration of monocytes. J Cell Biol 142:595–607
Weinacker AB, Vaszar LT (2001) Acute respiratory distress syndrome: physiology and new management strategies. Annu Rev Med 52:221–237
Zanetti A, Conforti G, Hess S, Martin-Padura I, Ghibaudi E, Preissner KT, Dejana E (1994) Clustering of vitronectin and RGD peptides on microspheres leads to engagement of integrins on the luminal aspect of endothelial cell membrane. Blood 84:1116–1123
Acknowledgements
This work was supported through grants from Natural Sciences and Engineering Research Council of Canada and Saskatchewan Health Research Foundation to Dr. Singh. Dr. Janardhan is supported through a University of Saskatchewan Graduate Merit Scholarship. We thank Ms Sarah Caldwell for help with immunogold electron microscopy.
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Janardhan, K.S., Appleyard, G.D. & Singh, B. Expression of integrin subunits αv and β3 in acute lung inflammation. Histochem Cell Biol 121, 383–390 (2004). https://doi.org/10.1007/s00418-004-0649-1
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DOI: https://doi.org/10.1007/s00418-004-0649-1