Extracellular Matrix and Bronchial Epithelial Cell Migration* K.A Rickard, I.D.;]. Taylor; B.S.;).R. 5.1. Rennard, t.D., F.G.G.P.
pur;::.em, M.D.' and
M
ib'Tation of pith Hal cells is an important ~ atur of wound healing. ompon nts of extracellular matrix are chemotactic for oth r cell, and w hypoth sized that bas ment m mbran compon nts, laminin and typ I hemotacti for bo in bronchial epith Hal collagen, ar c II (BBE ). Th BBEC w r obtain d ancl cultured for 3 r. Omaha.
clays in m c1ium 199 stlppl m nt d with insulin, tran f< rrin, hydrocortison, pidermal growth fa tor, antibiotics, and 10% fetal calf rum. Aft r rin ing, th II w r tryp iniz d and restlsp ncled at 1 X 1 in un uppl III nt d m dium 199, and used as th II sour f< r ch m taxi. hemotaxi was measur d by a modill d Boyden b(jnd-\ II lamb r technique using -l.l.1l1 filters ( ucleopor) coat d with 1'7l gelatin in a -multiwell cbamb r for 6 h at 37 Th onhalf maximal dos for migration was 10 IJ,glml for pIa ma fibr nectin, 1 l.l.glmI for typ I collag n, and 1 IJ,glml for laminin. " h ckerhoard" analysi d monstratcd that th migration was both chemokinetic and chemota tic for all th omponents tudied. To test if BBE attachm tra eHular matrix com0
•
IGUIlE 1. BB· chemotaxis w m ured by th blind-w II chamber t chniqu. hemotaclie a tivity \Va quantital d by L'Ounting COm pI lei)' migraled cells. h mota tic Ii It r II d wer photographed lind r X 200 magn i f!calioll. h motaxis to Ih folloy,~ng components is shown: A, Gbronectin on g lalin-coated filter; B, Abron tin on laminin-coal d filter; G. fibronectin on type I C'OlIagen-C'Oated filler; D, type r collagen on g la in-coated flit r; E. typ IV colla!! n On I:1minincoated IiIter; F. type r C'OlIagen on typ I (''Gllag n-coat d filt r; G, bmin'n on gelatin-coat d filter; H laminin on type I collag n-coat d filt r; and I, laminin on type I collagen-coated lilter.
CHEST' 101 , 3 I MARCH, 1992 , Supplement
175
ponents would have an effect on chemotaxis, the chemotactic filters were soaked in a solution of either type IV collagen, laminin, or type I collagen at a concentration of 10 ....Wml and ai~dried. Chemotaxis using a type IV collagen-coated filter was increased to fibronectin but greatly decreased to laminin. Chemotaxis using a type I collagen-coated filter was greatly increased to both type IV collagen and laminin. Finally, chemotaxis, using a laminin-coated filter, was decreased for both fibronectin and type IV collagen. Thus, it appears that type I collagen can greatly enhance the chemotactic migration of the BBEC, but that basement membrane components, in general, reduce the ability of these cells to migrate in response to matrix components (Fig 1). The ability of BBECs to migrate to specific stimuli has been demonstrated previously using the modified Boyden blind-well chemotactic chamber technique. I The recruitment of cells to an area in vivo likely requires that the cells
188
not only possess the ability to migrate, but may also depend on the cells ability to attach to the subjacent matrix. Thus, the composition of the matrix may have a dramatic effect on the chemotactic migration of airway epithelial cells. In summary, BBEC migration increases when placed on a type I collagen-coated filter. However, when a filter coated with the basement membrane components, laminin or type IV collagen, is used, the ability of the BBEC to migrate to connective tissue matrix components is reduced. This may play an important role in repair processes which involve injury to the bronchial epithelium, in particular, those injuries which may alter the composition of the basement membrane. REFERENCE 1 Shoji 5, Rickard KA, Ertl RF, Linder J, Rennard 51. Lung fibroblasts produce chemotactic factors for bronchial epithelial cells. Am J Physioll989; 257:71-79
34th Annual Thomas L Petty Aspen Lung Conference
pur;::.em, M.D.' and
M
ib'Tation of pith Hal cells is an important ~ atur of wound healing. ompon nts of extracellular matrix are chemotactic for oth r cell, and w hypoth sized that bas ment m mbran compon nts, laminin and typ I hemotacti for bo in bronchial epith Hal collagen, ar c II (BBE ). Th BBEC w r obtain d ancl cultured for 3 r. Omaha.
clays in m c1ium 199 stlppl m nt d with insulin, tran f< rrin, hydrocortison, pidermal growth fa tor, antibiotics, and 10% fetal calf rum. Aft r rin ing, th II w r tryp iniz d and restlsp ncled at 1 X 1 in un uppl III nt d m dium 199, and used as th II sour f< r ch m taxi. hemotaxi was measur d by a modill d Boyden b(jnd-\ II lamb r technique using -l.l.1l1 filters ( ucleopor) coat d with 1'7l gelatin in a -multiwell cbamb r for 6 h at 37 Th onhalf maximal dos for migration was 10 IJ,glml for pIa ma fibr nectin, 1 l.l.glmI for typ I collag n, and 1 IJ,glml for laminin. " h ckerhoard" analysi d monstratcd that th migration was both chemokinetic and chemota tic for all th omponents tudied. To test if BBE attachm tra eHular matrix com0
•
IGUIlE 1. BB· chemotaxis w m ured by th blind-w II chamber t chniqu. hemotaclie a tivity \Va quantital d by L'Ounting COm pI lei)' migraled cells. h mota tic Ii It r II d wer photographed lind r X 200 magn i f!calioll. h motaxis to Ih folloy,~ng components is shown: A, Gbronectin on g lalin-coated filter; B, Abron tin on laminin-coal d filter; G. fibronectin on type I C'OlIagen-C'Oated filler; D, type r collagen on g la in-coated flit r; E. typ IV colla!! n On I:1minincoated IiIter; F. type r C'OlIagen on typ I (''Gllag n-coat d filt r; G, bmin'n on gelatin-coat d filter; H laminin on type I collag n-coat d filt r; and I, laminin on type I collagen-coated lilter.
CHEST' 101 , 3 I MARCH, 1992 , Supplement
175
ponents would have an effect on chemotaxis, the chemotactic filters were soaked in a solution of either type IV collagen, laminin, or type I collagen at a concentration of 10 ....Wml and ai~dried. Chemotaxis using a type IV collagen-coated filter was increased to fibronectin but greatly decreased to laminin. Chemotaxis using a type I collagen-coated filter was greatly increased to both type IV collagen and laminin. Finally, chemotaxis, using a laminin-coated filter, was decreased for both fibronectin and type IV collagen. Thus, it appears that type I collagen can greatly enhance the chemotactic migration of the BBEC, but that basement membrane components, in general, reduce the ability of these cells to migrate in response to matrix components (Fig 1). The ability of BBECs to migrate to specific stimuli has been demonstrated previously using the modified Boyden blind-well chemotactic chamber technique. I The recruitment of cells to an area in vivo likely requires that the cells
188
not only possess the ability to migrate, but may also depend on the cells ability to attach to the subjacent matrix. Thus, the composition of the matrix may have a dramatic effect on the chemotactic migration of airway epithelial cells. In summary, BBEC migration increases when placed on a type I collagen-coated filter. However, when a filter coated with the basement membrane components, laminin or type IV collagen, is used, the ability of the BBEC to migrate to connective tissue matrix components is reduced. This may play an important role in repair processes which involve injury to the bronchial epithelium, in particular, those injuries which may alter the composition of the basement membrane. REFERENCE 1 Shoji 5, Rickard KA, Ertl RF, Linder J, Rennard 51. Lung fibroblasts produce chemotactic factors for bronchial epithelial cells. Am J Physioll989; 257:71-79
34th Annual Thomas L Petty Aspen Lung Conference
