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Intranasal delivery of the cytoplasmic domain of CTLA-4 using a novel protein transduction domain prevents allergic inflammation

Nature Medicine volume 12, pages 574–579 (2006)Cite this article

Abstract

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.

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Figure 1: Evaluation of Hph-1 as a novel protein-transduction domain.
Figure 2: Inhibitory effect of Hph-1–ctCTLA-4 on T-cell activation.
Figure 3: In vivo transduction of Hph-1–ctCTLA-4-EGFP through intranasal route.
Figure 4: Therapeutic effect of nasal administration of Hph-1–ctCTLA-4 on airway inflammation in OVA-induced experimental mouse model of asthma.

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Acknowledgements

We thank A. Bothwell and T. Morio for the comments on the manuscript and all members of Immune Cell Engineering Laboratory, Department of Biotechnology, Yonsei University for their spiritual help. This work was supported in part by research grants to S.K.L. from Korea Institute of Industrial Technology Evaluation and Planning (M1-0310-40-0000), Korea Health Industry Development Institute (0412-DB00-0101-0011), Korea Science and Engineering Foundation (2005-00117) and Korea Rural Economic Institute (204081-3). This work was also supported in part by research grants to C.S.P. from the Korea Health 21 R&D Project, Ministry of Health and Welfare, Republic of Korea (01-PJ3-PG6-01GN04-003).

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Authors and Affiliations

  1. Department of Biotechnology, College of Engineering, Yonsei University, Seoul, 120-749, Republic of Korea

    Je-Min Choi, Wook-Jin Chae, Yung-Gook Jung, Jae-Chul Park, Kyun-Do Kim, Eun-Sung Kim & Sang-Kyou Lee

  2. Genome Research Center for Allergy and Respiratory Disease, Soonchunhyang University Hospital, Bucheon, 420-767, Republic of Korea

    Mi-Hyun Ahn, Hyun-Mi Song, Young-Eun Kim, Jung-Ah Shin & Choon-Sik Park

  3. Department of Internal Medicine, Division of Allergy and Immunology, Yonsei Medical School, Seoul, 120-749, Republic of Korea

    Jung-Won Park

  4. Department of Ophthalmology, Soonchunhyang University Hospital, Bucheon, 420-767, Republic of Korea

    Tae-Kwann Park

  5. Department of Dermatology, Chungnam National University Hospital, Daesa-dong, Jung-gu, Daejon, 301-721, Republic of Korea

    Jung-Hoon Lee

  6. Skin Research Institute, Amorepacific Co., R&D Center, Yongin, 449-729, Republic of Korea

    Byung-Fhy Seo

  7. ForHumanTech Co., Ltd, Kowoon Institute of Technology Innovation Bldg 706, Suwon, Republic of Korea

    Dong-Ho Lee, Seung-Kyou Lee & Sang-Kyou Lee

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Correspondence to Sang-Kyou Lee.

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Supplementary information

Supplementary Fig. 1

Concentration-dependent transduction of Hph-1–PTD conjugated β-gal in vivo. (PDF 78 kb)

Supplementary Fig. 2

In vivo toxicity of Hph-1–ctCTLA-4. (PDF 460 kb)

Supplementary Fig. 3

Immunogenecity of Hph-1–ctCTLA-4. (PDF 767 kb)

Supplementary Fig. 4

Inhibition of airway inflammation by Hph-1–ctCTLA-4 in concentration dependent manner. (PDF 1257 kb)

Supplementary Fig. 5

In vivo stability of Hph-1–ctCTLA-4 and its duration of anti-inflammatory activity. (PDF 1556 kb)

Supplementary Fig. 6

Effect of nasal administration of Hph-1–ctCTLA-4 on airway inflammation in TDI-induced experimental animal model for asthma. (PDF 1302 kb)

Supplementary Table 1

Blood biochemical test of Hph-1–ctCTLA-4 treated mice. (PDF 30 kb)

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Choi, JM., Ahn, MH., Chae, WJ. et al. Intranasal delivery of the cytoplasmic domain of CTLA-4 using a novel protein transduction domain prevents allergic inflammation. Nat Med 12, 574–579 (2006). https://doi.org/10.1038/nm1385

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