Characterization of Bovine Serum Factor Triggering the Lysis of Liposomes via Complement Activation
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概要
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Our previous studies have shown that the degree of damage to a liposome corresponds to the variability of the animal species from which the serum comes, and that a complement activating factor (CAF) plays an important role in inducing the activation of the complement system, ultimately leading to the lysis of the liposomes. In this study, our attention focused on the characterization of the bovine serum factor (bCAF) that is involved in complement-mediated immune lysis of the liposome. The active fraction containing CAF partially purified with PEG and ammonium sulfate results in marked activation of the complement system via the alternative pathway when interacted with CAF-depleted serum, whereas the active fraction or CAF-depleted serum alone does not activate the complement. The interaction between lipopolysaccharide (LPS), heparin, zymosan or their mixture in place of CAF and CAF-depleted serum does not result in any significant activation of the complement system. Results from pretreatment with rabbit anti-bovine IgM IgG and rabbit anti-bovine IgG IgG indicate that activation of the complement system is not attributable to the antibody which is generally involved in activation of complement via the classical pathway. The results have further been proven by pretreatment with Concanavalin A (Con A) sepharose and protein G sepharose ruling out the possibility of antibody-mediated activation of complement. Our studies on collagenase and trypsin digestion demonstrate that the relative activity of CAF does not diminish with increase in collagenase concentration, and decreases with increase in trypsin concentration, strongly indicating that CAF does not have a collagen-like domain in its structure. The relative activity of CAF is dramatically inhibited after reduction with 2-mercaptoethanol (2-ME), clearly demonstrating that CAF is sensitive to reduction with 2-ME and confirming a sulhydryl-dependent protein. The optimal activity of CAF is observed in the range of 35-45℃ and its half-life at 37℃ is about 105 h. Furthermore, the relative activity of DAF increases and gradually approaches a plateau level with the increase of Mg^<2+> concentration. Obviously, complement activation induced by CAF depends on adequate Mg^<2+> concentration, confirming that this dependence is characterisitic of the altenative pathway.
- 公益社団法人日本薬学会の論文
- 1998-04-15
著者
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Kiwada Hiroshi
徳島大学 ヘルスバイオサイエンス研究部薬物動態制御学
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Kiwada Hiroshi
Faculty Of Pharmaceutical Science University Of Tokyo:(present Address)faculty Of Pharmaceutical Sci
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Kiwada Hiroshi
Faculty Of Pharmaceutifal Sciences The University Of Tokushima
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Higashisaka Kazuma
Dep. Of Toxicology And Safety Sci. Graduate School Of Pharmaceutical Sciences Osaka Univ.
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Kamada Haruhiko
医薬基盤研究所 創薬プロテオミクスプロジェクト
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Kamada Haruhiko
Laboratory Of Pharmaceutical Proteomics (lpp) National Institute Of Biomedical Innovation (nibio)
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Kiwada Hiroshi
Department Of Pharmacokinetics And Biopharmaceutics Faculty Of Pharmaceutical Sciences The Universit
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Kamiya Hiroyuki
Lab. For Molecular Design Of Pharmaceutics Fac. Of Pharmaceutical Sciences Hokkaido Univ.
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Kamada H
Yamagata Technopolis Foundation Yamagata Jpn
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Kamada Hitoshi
Institute For Life Support Technology Yamagata Promotional Organization For Industrial Technology
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ISHIDA Tatsuhiro
Faculty of Pharmaceutical Sciences, The University of Tokushima
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LIU Shicheng
Faculty of Pharmaceutical Sciences, The University of Tokushima
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Liu Shicheng
Faculty Of Pharmaceutical Sciences The University Of Tokushima
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Kamada H
Institute For Life Support Technology Yamagata Promotional Organization For Industrial Technology
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Kiwada H
Faculty Of Pharmaceutifal Sciences The University Of Tokushima
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Kiwada Hiroshi
Faculty Of Pharmaceutical Science The University Of Tokushima
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Kiwada Hiroshi
Department Of Pharmacokinetics And Biopharmaceutics Subdivision Of Biopharmaceutical Science Institu
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Ishida Tatsuhiro
Dep. Of Pharmacokinetics And Biopharmaceutics Subdivision Of Biopharmaceutical Sciences Inst. Of Hea
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Ishida Tatsuhiro
Department Of Pharmacokinetics And Biopharmaceutics Subdivision Of Biopharmaceutical Science Institu
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