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Engineering N-glycosylation mutations in IL-12 enhances sustained cytotoxic T lymphocyte responses for DNA immunization

Nature Biotechnology volume 20, pages 381–386 (2002)Cite this article

Abstract

Interleukin-12 (IL-12), consisting of p40 and p35 subunits, produces both p70 heterodimer and free p40. p70 is essential for the induction of T-helper 1 (Th1) and cytotoxic T-cell (CTL) immunity, whereas p40 inhibits p70-mediated function. Here, we found that mutations introduced into N-glycosylation sites (N220 of murine p40 and N222 of human p40) reduced secretion of p40 but not p70. Co-immunization of N220 mutant mIL-12 gene with hepatitis C virus (HCV) E2 DNA significantly enhanced long-term E2-specific CD8+ T-cell response and protection against tumor challenge compared with that of wild type. Our results indicate that the ratio of p70 to p40 is important for generating sustained long-term cell-mediated immunity. Thus, the mutant IL-12 could be utilized for the development of DNA vaccines as an adjuvant for the generation of long-term memory T-cell responses.

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Figure 1: Western blot analysis of human p40 and its derivatives with mutations at putative N-glycosylation sites.
Figure 2: Comparison of receptor binding and IFN-γ induction between wild-type and N222L mutant p70 on Con A blast cells.
Figure 3: Kinetics of E2-specific antibody responses in DNA immunization.
Figure 4: Effect of mIL-12N220L on E2-specific CTL responses and protection against challenge with modified tumors.

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Acknowledgements

The authors acknowledge Sang-Chun Lee and Su-Il Park for devoted animal care. We are grateful to Hye-Ryun Kim and Se-Hwan Yang for faithful reading and comments. This work was supported by National Research Laboratory grant from Korea Institute of Science and Technology Evaluation and Planning (2000-N-NL-01-C-202) and grants from POSCO (2000Y013), Superior Research Center supported by Korea Science and Engineering Foundation, and ProGen Co. Ltd.

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  1. Sang J. Ha and Jun Chang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Life Science, National Research Laboratory of DNA Medicine, Pohang University of Science & Technology, San 31, Hyoja-Dong, Nam-Ku, Pohang, 790-784, Kyungbuk, Korea

    Sang J. Ha, Jun Chang, Man K. Song, You S. Suh, Hyun T. Jin, Chu H. Lee, Gyu H. Nam, Gildon Choi, Kwan Y. Choi & Young C. Sung

  2. ProGen Co. Ltd., 47-5, Sanggal-Ri, Kiheung-Up, Youngin, 449-905, Kyunggi, Korea

    Sung H. Lee & Won B. Kim

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  1. Sang J. Ha
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Correspondence to Young C. Sung.

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Ha, S., Chang, J., Song, M. et al. Engineering N-glycosylation mutations in IL-12 enhances sustained cytotoxic T lymphocyte responses for DNA immunization. Nat Biotechnol 20, 381–386 (2002). https://doi.org/10.1038/nbt0402-381

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