Published ahead of print on August 1, 2003, doi:10.1165/rcmb.2003-0198OC Am. J. Respir. Cell Mol. Biol., Volume 30, Number 2, February 2004, 166-173 A more recent version of this article appeared on February 1, 2004
Submitted on May 27, 2003 Heterogeneity of heparan sulfates in human lungNicole C Smits1,1 Biochemistry, University Medical Center Nijmegen, Nijmegen, The Netherlands, 2 Pulmonary Diseases, University Medical Center Nijmegen, Nijmegen, The Netherlands * To whom correspondence should be addressed. E-mail: a.vankuppevelt{at}ncmls.kun.nl.
Heparan sulfates (HS), a class of glycosaminoglycans, are long linear complex polysaccharides covalently attached to a protein core. The HS molecules are made up of repeating disaccharides onto which modification patterns are superimposed. This results in a large structural heterogeneity and forms the basis of specific interactions of HS towards a vast array of proteins, including growth factors and proteases. To study HS heterogeneity in the lung, we used phage display technology to select seven antibodies against human lung HS. Antibodies reacted with HS/heparin, but not with other glycosaminoglycans or polyanions. Sulfate groups were essential for antibody binding. The amino acid sequence of the antibodies was established, the complementarity determining region 3 of the heavy chain containing basic amino acids. The antibodies defined HS epitopes with a characteristic tissue distribution. Antibody EV3A1 primarily stained macrophages. Other antibodies primarily stained basement membranes, but with different preference towards type of basement membrane. Antibody EV3C3 was the only antibody which clearly reacted with bronchiolar epithelial cells. In human lung parenchyma bFGF and VEGF were largely bound by HS. Some antibodies blocked a bFGF-binding site of HS, and one antibody blocked a VEGF-binding site of heparin. Taken together, these data suggest a specific role for HS epitopes in human lung. The antibodies obtained may be valuable tools to study HS in pulmonary diseases.
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