Effect of cholesterol content in activation of the classical versus the alternative pathway of rat complement system induced by hydrogenated egg phosphatidylcholine-based liposomes.
Liposomes composed of hydrogenated egg phosphatidylcholine (HEPC) and cholesterol (CHOL) were found to activate the rat complement (C) system in a CHOL content-dependent manner. Liposomes containing 22 or 33 mol% CHOL activated the C system in a Ca(2+)-dependent manner, suggesting that C activation occurred via the classical pathway. Liposomes containing 44 mol% CHOL activated the C system in a Ca(2+) independent manner, suggesting that C activation occurred via the alternative pathway. The CHOL content appeared to dictate the pathway by which the C system was activated. This C activation was inhibited by removal of serum component(s), which adsorb to the liposomes. Activation of the alternative pathway, induced by the liposomes, was reduced by the depletion of IgG and IgM, whereas the classical pathway activation was reduced by the depletion of IgG, but not IgM. In addition, the removal of adsorbed serum component(s) by treatment with 44 mol% CHOL-containing liposomes decreased serum IgG and IgM levels that adsorb to the same liposomes, whereas the removal of adsorbed serum component(s) by treatment with 22 mol% CHOL-containing liposomes only slightly decreased serum IgG levels, which adsorbs to the same liposomes. Collectively, both IgG and IgM, which are specifically adsorbed to the liposomes in a CHOL-content dependent manner, were responsible for C activation via the alternative pathway induced by the 44 mol% CHOL containing liposomes. IgG alone would be partially responsible for C activation via the classical pathway induced by 22 or 33 mol% CHOL-containing liposomes. The discovery of this unique C-activating property of liposomes will be of value in attempts to decipher the underlying mechanism of C activation by providing a useful model membrane system.
Ishida
T
T
Department of Pharmacokinetics, Graduate School of Pharmaceutical Sciences, The University of Tokushima, 1-78-1 Sho-machi, Tokushima 770-8505, Japan.
Yasukawa
K
K
Kojima
H
H
Harashima
H
H
Kiwada
H
H
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D002784:Cholesterol / Q000032:analysis*; D003170:Complement Pathway, Alternative; D003171:Complement Pathway, Classical; D004797:Enzyme-Linked Immunosorbent Assay; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000378:metabolism; D007075:Immunoglobulin M / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry; D008297:Male; D010755:Organophosphates / Q000378:metabolism; D010713:Phosphatidylcholines / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar
D001798:Blood Proteins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008081:Liposomes; D010755:Organophosphates; D010713:Phosphatidylcholines; C002290:dicetylphosphate; D002784:Cholesterol