title: Neutralization of leukotriene C4 and D4 activity by monoclonal and single-chain antibodies.
abstract: BACKGROUND
  Cysteinyl leukotrienes (LTs) are key mediators in inflammation. To explore the structure of the antigen-recognition site of a monoclonal antibody against LTC4 (mAbLTC), we previously isolated full-length cDNAs for heavy and light chains of the antibody and prepared a single-chain antibody comprising variable regions of these two chains (scFvLTC).
  
  
  METHODS
  We examined whether mAbLTC and scFvLTC neutralized the biological activities of LTC4 and LTD4 by competing their binding to their receptors.
  
  
  RESULTS
  mAbLTC and scFvLTC inhibited their binding of LTC4 or LTD4 to CysLT1 receptor (CysLT1R) and CysLT2 receptor (CysLT2R) overexpressed in Chinese hamster ovary cells. The induction by LTD4 of monocyte chemoattractant protein-1 and interleukin-8 mRNAs in human monocytic leukemia THP-1 cells expressing CysLT1R was dose-dependently suppressed not only by mAbLTC but also by scFvLTC. LTC4- and LTD4-induced aggregation of mouse platelets expressing CysLT2R was dose-dependently suppressed by either mAbLTC or scFvLTC. Administration of mAbLTC reduced pulmonary eosinophil infiltration and goblet cell hyperplasia observed in a murine model of asthma. Furthermore, mAbLTC bound to CysLT2R antagonists but not to CysLT1R antagonists.
  
  
  CONCLUSIONS
  These results indicate that mAbLTC and scFvLTC neutralize the biological activities of LTs by competing their binding to CysLT1R and CysLT2R. Furthermore, the binding of cysteinyl LT receptor antagonists to mAbLTC suggests the structural resemblance of the LT-recognition site of the antibody to that of these receptors.
  
  
  CONCLUSIONS
  mAbLTC can be used in the treatment of inflammatory diseases such as asthma.
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kawakami
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Shiori
authors.initials: S
authors.lastname: Hirano
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Mai
authors.initials: M
authors.lastname: Kinoshita
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Akemi
authors.initials: A
authors.lastname: Otsuki
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Toshiko
authors.initials: T
authors.lastname: Suzuki-Yamamoto
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Masumi
authors.initials: M
authors.lastname: Kimoto
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Sae
authors.initials: S
authors.lastname: Sasabe
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Fukushima
authors.affiliation: Department of Biochemistry, Gunma University Graduate School of Medicine, Gunma 371-8511, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Kishimoto
authors.affiliation: Department of Biochemistry, Gunma University Graduate School of Medicine, Gunma 371-8511, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Izumi
authors.affiliation: Department of Respiratory Care & Sleep Control Medicine, Kyoto University Graduate School of Medicine, Kyoto 606-8501, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Oga
authors.affiliation: Department of Pharmacology, Kyoto University Graduate School of Medicine, Kyoto 606-8501, Japan.
authors.forename: Shuh
authors.initials: S
authors.lastname: Narumiya
authors.affiliation: Structural Biophysics Laboratory, RIKEN SPring-8 Center, Harima Institute, Hyogo 679-5148, Japan.
authors.forename: Mitsuaki
authors.initials: M
authors.lastname: Sugahara
authors.affiliation: Structural Biophysics Laboratory, RIKEN SPring-8 Center, Harima Institute, Hyogo 679-5148, Japan; Department of Chemistry and Biological Science, College of Science and Engineering, Aoyama Gakuin University, Kanagawa 252-5258, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Miyano
authors.affiliation: Department of Food and Nutrition, Kyoto Women's University, Kyoto 605-8501, Japan.
authors.forename: Shozo
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Okayama 719-1197, Japan. Electronic address: ytaka@fhw.oka-pu.ac.jp.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Takahashi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology* / Q000627:therapeutic use; D001249:Asthma / Q000188:drug therapy; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D016207:Cytokines / Q000096:biosynthesis; D006801:Humans; D020024:Leukotriene Antagonists / Q000494:pharmacology; D017997:Leukotriene C4 / Q000276:immunology*; D017998:Leukotriene D4 / Q000276:immunology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010974:Platelet Aggregation / Q000187:drug effects; D018077:Receptors, Leukotriene / Q000187:drug effects / Q000502:physiology; D057127:Single-Chain Antibodies / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D016207:Cytokines; D020024:Leukotriene Antagonists; D018077:Receptors, Leukotriene; D057127:Single-Chain Antibodies; D017997:Leukotriene C4; D017998:Leukotriene D4; C412375:cysteinyl leukotriene receptor 2
keywords: Arachidonic acid; CysLT(1) receptor; CysLT(2) receptor; Cysteinyl leukotriene; Monoclonal antibody; Single-chain variable fragment