Enzyme studies on Epstein-Barr virus-transformed lymphoid cell lines from Wolman's disease: Lipases, cholesterol esterase and 4-methylumbelliferyl acyl ester hydrolases

Abstract

(1) In lymphoid cell lines established by Epstein-Barr virus transformation of B-lymphocytes from normal subjects there exist two lipases hydrolysing triolein (the first one with acid optimum pH and the other one with alkaline optimum pH) and one cholesterol esterase (with acidic optimum pH). The acid triolein lipase (optimum pH 3.75–4.0) and the acid cholesterol esterase are activated by taurocholate (optimal concentration between 1 and 2.5 g/l) whereas alkaline triolein-lipase is inhibited by crude taurocholate. (2) Acid lipase deficiency is demonstrated in lymphoid cell lines from a Wolman's patient, using natural substrates, triolein and cholesteryl oleate (residual activity 5 and 8%, respectively). Thus, this similar deficiency demonstrates that, in lymphoid cell lines, triolein and cholesteryl esters are hydrolysed (under the conditions used here) by a single enzyme i.e., lysosomal acid lipase muted in Wolman's disease. (3) pH profiles of synthetic substrate hydrolysis show marked differences between methylumbelliferyl oleate and methylumbelliferyl palmitate, and are greatly dependent on the assay conditions used. In the presence of optimal concentrations of taurocholate (1–2.5 g/l), nonspecific carboxylesterases are inhibited and acid lipase is activated: in this case, methyl-umbelliferyl oleate can be used to demonstrate the acid lipase deficiency in Wolman's lines (15–20% of residual activity). Methylumbelliferyl palmitate hydrolysis is less dependent on assay conditions and thus can be more accurately used for the diagnosis of Wolman's disease, with lower residual activity (10–15%) than using methylumbelliferyl oleate. Thus, Epstein-Barr virus-transformed lymphoid cell lines represent an accurate model system in culture for experimental studies of Wolman's disease.