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Immunocytochemical localization of latent transforming growth factor-beta1 activation by stimulated macrophages.

摘要:

Transforming growth factor-beta1 (TGF-beta) is secreted in a latent form consisting of mature TGF-beta noncovalently associated with its amino-terminal propeptide, which is called latency associated peptide (LAP). Biological activity depends upon the release of TGF-beta from the latent complex following extracellular activation, which appears to be the key regulatory mechanism controlling TGF-beta action. We have identified two events associated with latent TGF-beta (LTGF-beta) activation in vivo: increased immunoreactivity of certain antibodies that specifically detect TGF-beta concomitant with decreased immunoreactivity of antibodies to LAP. Macrophages stimulated in vitro with interferon-gamma and lipopolysaccharide reportedly activate LTGF-beta via cell membrane-bound protease activity. We show through dual immunostaining of paraformaldehyde-fixed macrophages that such physiological TGF-beta activation is accompanied by a loss of LAP immunoreactivity with concomitant revelation of TGF-beta epitopes. The induction of TGF-beta immunoreactivity colocalized with immunoreactive betaglycan/RIII in activated macrophages, suggesting that LTGF-beta activation occurs on the cell surface. Confocal microscopy of metabolically active macrophages incubated with antibodies to TGF-beta and betaglycan/RIII prior to fixation supported the localization of activation to the cell surface. The ability to specifically detect and localize LTGF-beta activation provides an important tool for studies of its regulation.

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作者: H,Chong [1] ; Y,Vodovotz [2] ; G W,Cox [3] ; M H,Barcellos-Hoff [4]
作者单位: Life Sciences Division [1] Lawrence Berkeley National Laboratory [2] University of California [3] Berkeley 94720 [4] USA. [5]
DOI: 10.1002/(SICI)1097-4652(199903)178:3<275::AID-JCP1>3.0.CO;2-Q
PMID: 9989773
发布时间: 2006-11-15
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