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.

GENERAL SIGNIFICANCE:

mAbLTC can be used in the treatment of inflammatory diseases such as asthma.

• Authors: Yuki Kawakami, Shiori Hirano, Mai Kinoshita, Akemi Otsuki, Toshiko Suzuki-Yamamoto, Makiko Suzuki, Masumi Kimoto, Sae Sasabe, Mitsuo Fukushima, Koji Kishimoto, Takashi Izumi, Toru Oga, Shuh Narumiya, Mitsuaki Sugahara, Masashi Miyano, Shozo Yamamoto, Yoshitaka Takahashi
• Journal: Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 1840 Issue 6 Pg. 1625-33 (Jun 2014) ISSN: 0006-3002 [Print] Netherlands
• PMID: 24361619 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier B.V. All rights reserved.

Chemical References

• Antibodies, Monoclonal
• Cytokines
• Leukotriene Antagonists
• Receptors, Leukotriene
• Single-Chain Antibodies
• Leukotriene C4
• Leukotriene D4
• cysteinyl leukotriene receptor 2

Topics

• Animals
• Antibodies, Monoclonal (pharmacology, therapeutic use)
• Asthma (drug therapy)
• CHO Cells
• Cricetinae
• Cricetulus
• Cytokines (biosynthesis)
• Humans
• Leukotriene Antagonists (pharmacology)
• Leukotriene C4 (immunology)
• Leukotriene D4 (immunology)
• Male
• Mice
• Mice, Inbred C57BL
• Platelet Aggregation (drug effects)
• Receptors, Leukotriene (drug effects, physiology)
• Single-Chain Antibodies (pharmacology)