R Osuský1, S M Walker, S J Ryan. 1. Department of Ophthalmology, School of Medicine, University of Southern California, Los Angeles, USA.
Abstract
BACKGROUND: Macrophages play an important role in several ocular diseases. Because macrophages localized in ocular tissues may be derived from blood monocytes, the effect of vitreous [containing transforming growth factor-beta 2 (TGF-beta 2) and hyaluronic acid] on blood monocytes, maturating in the tissue to macrophages, was determined. METHODS: Human monocytes were cultured with and without vitreous in RPMI 1640 medium containing human AB serum. As a parameter of activation the release of interleukin-6 was measured by the B9 bioassay; as an indication of maturation, the content of acid phosphatase and the increase in cell size were assessed. RESULTS: Monocytes in vitreous-containing medium grew more slowly than did control monocytes. Monocytes cultured in 10% vitreous released 51% less, and in 20% vitreous 73% less, interleukin-6 than control monocytes. Vitreous at 20% significantly (P = 0.0075) reduced the amount of acid phosphatase by 80% over a 4-day culture period. This reduction was partially eliminated with neutralizing antibodies to TGF-beta (P = 0.0014). Furthermore, human recombinant TGF-beta 2 increased the activity of acid phosphatase in monocytes at 1.25 ng/ml and reduced it (P < 0.0001) at higher concentrations (5-10 ng/ml). Hyaluronic acid showed an effect additive to that of TGF-beta in further diminishing the amount of acid phosphatase (P = 0.026). CONCLUSION: Vitreous exerts a regulatory effect on monocyte activation and maturation by its content of TGF-beta and possibly hyaluronic acid and may, thus, modify the inflammatory or immune response in the eye.
BACKGROUND: Macrophages play an important role in several ocular diseases. Because macrophages localized in ocular tissues may be derived from blood monocytes, the effect of vitreous [containing transforming growth factor-beta 2 (TGF-beta 2) and hyaluronic acid] on blood monocytes, maturating in the tissue to macrophages, was determined. METHODS:Human monocytes were cultured with and without vitreous in RPMI 1640 medium containing human AB serum. As a parameter of activation the release of interleukin-6 was measured by the B9 bioassay; as an indication of maturation, the content of acid phosphatase and the increase in cell size were assessed. RESULTS: Monocytes in vitreous-containing medium grew more slowly than did control monocytes. Monocytes cultured in 10% vitreous released 51% less, and in 20% vitreous 73% less, interleukin-6 than control monocytes. Vitreous at 20% significantly (P = 0.0075) reduced the amount of acid phosphatase by 80% over a 4-day culture period. This reduction was partially eliminated with neutralizing antibodies to TGF-beta (P = 0.0014). Furthermore, human recombinant TGF-beta 2 increased the activity of acid phosphatase in monocytes at 1.25 ng/ml and reduced it (P < 0.0001) at higher concentrations (5-10 ng/ml). Hyaluronic acid showed an effect additive to that of TGF-beta in further diminishing the amount of acid phosphatase (P = 0.026). CONCLUSION: Vitreous exerts a regulatory effect on monocyte activation and maturation by its content of TGF-beta and possibly hyaluronic acid and may, thus, modify the inflammatory or immune response in the eye.
Authors: Yuqin Yin; Qi Cui; Hui-Ya Gilbert; Yang Yang; Zhiyong Yang; Cynthia Berlinicke; Zhiwei Li; Camila Zaverucha-do-Valle; Huamei He; Victoria Petkova; Donald J Zack; Larry I Benowitz Journal: Proc Natl Acad Sci U S A Date: 2009-10-29 Impact factor: 11.205