Regulation of the effects of TGF-beta 1 by activation of latent TGF-beta 1and differential expression of TGF-beta receptors (T beta R-I and T beta R-II) in idiopathic pulmonary fibrosis

Background-Idiopathic pulmonary fibrosis (IPF) is characterised by subpleur al fibrosis that progresses to involve all areas of the lung. The expressio n of transforming growth factor-beta1. (TGF-beta1), a potent regulator of c onnective tissue synthesis, is increased in lung sections of patients with IPF. TGF-beta1 is generally released in a biologically latent form (L-TGF-b eta1). Before being biologically active, TGF-P must be converted to its act ive form and interact with both TGF-beta receptors type I and II (T betaR-I and T betaR-II). TGF-beta latency binding protein 1 (LTBP-1), which facili tates the release and activation of L-TGF-beta1, is also important in the b iology of TGF-beta1. Methods-Open lung biopsy samples from patients with IPF and normal controls were examined to localise T betaR-I, T betaR-II, and LTBP-1. Alveolar macr ophages (AM) and bronchoalveolar lavage (BAL) fluid were examined using the CCL-64 bioassay to determine if TGF-beta is present in its active form in the lungs of patients with IPF. Results-Immunoreactive L-TGF-beta1 was present in all lung cells of patient s with IPF except for fibroblasts in the subepithelial regions of honeycomb cysts. LTBP-1 was detected primarily in AM and epithelial cells lining hon eycomb cysts in areas of advanced IPF. In normal lungs LTBP-1 immunoreactiv ity was observed in a few AM. AM from the upper and lower lobes of patients with IPF secreted 1.6 (0.6) fmol and 4.1 (1.9) fmol active TGF-beta, respe ctively, while AM from the lower lobes of control patients secreted no acti ve TGF-beta (p less than or equal to0.01 for TGF-beta in the conditioned me dia from AM obtained from the lower lobes of IPF patients v normal controls ). The difference in percentage active TGF-beta secreted by AM from the low er lobes of patients with IPF and the lower lobes of control patients was s ignificant (p less than or equal to0.01), but the difference between the to tal TGF-beta secreted from these lobes was not significant. The difference in active TGF-beta in conditioned media of AM from the upper and lower lobe s of patients with IPF was also not statistically significant. BAL fluid fr om the upper and lower lobes of patients with IPF contained 0.7 (0.2) fmol and 2.9 (1.2) fmol active TGF-beta, respectively (p less than or equal to0. 03). The percentage of active TGF-beta in the upper and lower lobes was 17. 6 (1.0)% and 78.4 (1.6)%, respectively (p less than or equal to0.03). In co ntrast, BAL fluid from control patients contained small amounts of L-TGF-be ta. Using immunostaining, both T betaR-I and T betaR-II were present on all cells of normal lungs but T betaR-I was markedly reduced in most cells in areas of honeycomb cysts except for interstitial myofibroblasts in lungs of patients with IPF. TGF-beta1 inhibits epithelial cell proliferation and a lack of T betaR-I expression by epithelial cells lining honeycomb cysts wou ld facilitate repair of the alveoli by epithelial cell proliferation. Howev er, the presence of both T beta Rs on fibroblasts is likely to result in a response to TGF-beta1 for synthesis of connective tissue proteins. Our find ings show that biologically active TGF-beta1 is only present in the lungs o f patients with IPF. In addition, the effects of TGF-beta1 on cells may be further regulated by the expression of T beta Rs. Conclusion-Activation of L-TGF-beta1 and the differential expression of T b eta Rs may be important in the pathogenesis of remodelling and fibrosis in IPF.