Intranasal delivery of the cytoplasmic domain of CTLA-4 using a novel protein transduction domain prevents allergic inflammation
Je-Min Choi1, Mi-Hyun Ahn2, Wook-Jin Chae1, Yung-Gook Jung1, Jae-Chul Park1, Hyun-Mi Song2, Young-Eun Kim2, Jung-Ah Shin2, Choon-Sik Park2, Jung-Won Park3, Tae-Kwann Park4, Jung-Hoon Lee5, Byung-Fhy Seo6, Kyun-Do Kim1, Eun-Sung Kim1, Dong-Ho Lee7, Seung-Kyou Lee7 & Sang-Kyou Lee1, 7
1 Department of Biotechnology, College of Engineering, Yonsei University, Seoul 120-749, Republic of Korea.
2 Genome Research Center for Allergy and Respiratory Disease, Soonchunhyang University Hospital, Bucheon, 420-767, Republic of Korea.
3 Department of Internal Medicine, Division of Allergy and Immunology, Yonsei Medical School, Seoul 120-749, Republic of Korea.
4 Department of Ophthalmology, Soonchunhyang University Hospital, Bucheon 420-767, Republic of Korea.
5 Department of Dermatology, Chungnam National University Hospital, Daesa-dong, Jung-gu, Daejon, 301-721, Republic of Korea.
6 Skin Research Institute, Amorepacific Co., R&D Center, Yongin 449-729, Republic of Korea.
7 ForHumanTech Co., Ltd, Kowoon Institute of Technology Innovation Bldg 706, Suwon, Republic of Korea.
CTLA-4 is a negative regulator of T-cell activation, and its inhibitory effects can be accomplished either by competition with CD28 or by transmitting negative signals through its intracellular domain. To utilize the cytoplasmic domain of CTLA-4 to suppress allergic inflammation, we fused it to a novel protein-transduction domain in the human transcriptional factor Hph-1. Transduction efficiency was verified in vitro and in vivo after ocular, intranasal and intradermal administration. After transduction into T cells, the Hph-1-ctCTLA-4 fusion protein inhibited the production of interleukin (IL)-2, and downregulated CD69 and CD25. Intranasal administration of Hph-1-ctCTLA-4 resulted in markedly reduced infiltration of inflammatory cells, secretion of T helper type 2 (TH2) cytokines, serum IgE levels and airway hyper-responsiveness in a mouse model of allergic airway inflammation. These results indicated that Hph-1-ctCTLA-4 constitutes an effective immunosuppressive protein drug for potential use in the treatment of allergic asthma, via nasal administration.