Neutralization of leukotriene C-4 and D-4 activity by monoclonal and single-chain antibodies

作者：Kawakami Yuki; Hirano Shiori; Kinoshita Mai; Otsuki Akemi; Suzuki Yamamoto Toshiko; Suzuki Makiko; Kimoto Masumi; Sasabe Sae; Fukushima Mitsuo; Kishimoto Koji; Izumi Takashi; Oga Toru; Narumiya Shuh; Sugahara Mitsuaki; Miyano Masashi; Yamamoto Shozo; Takahashi Yoshitaka^*

来源：Biochimica et Biophysica Acta-General Subjects, 2014, 1840(6): 1625-1633.

DOI：10.1016/j.bbagen.2013.12.016

• Arachidonic acid
• CysLI(1) receptor
• CysLT(2) receptor
• Cysteinyl leulcotriene
• Monoclonal antibody
• Single-chain variable fragment

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 mAbLIC 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 CysLT(1) receptor (CysLTiR) and Cy5LT(2) receptor (Cy5LT(2)R) 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 CysLIIR was dose-dependently suppressed not only by mAbLTC but also by scFvLTC. LTC4- and LTD4-induced aggregation of mouse platelets expressing Cy5LT(2)R 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 CysLT(2)R antagonists but not to CysLT(1)R antagonists. Conclusions: These results indicate that mAbLTC and scFvLTC neutralize the biological activities of LTs by competing their binding to CysLT(1)R and CysLT(2)R. 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.

• 出版日期2014-6
• 单位RIKEN