Engineering N-glycosylation mutations in IL-12 enhances sustained cytotoxic T lymphocyte responses for DNA immunization.
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.
Ha
Sang J
SJ
National Research Laboratory of DNA Medicine, Department of Life Science, Pohang University of Science & Technology, San 31, Hyoja-Dong, Nam-Ku, Pohang, Kyungbuk 790-784, Korea.
Chang
Jun
J
Song
Man K
MK
Suh
You S
YS
Jin
Hyun T
HT
Lee
Chu H
CH
Nam
Gyu H
GH
Choi
Gildon
G
Choi
Kwan Y
KY
Lee
Sung H
SH
Kim
Won B
WB
Sung
Young C
YC
D000818:Animals; D015153:Blotting, Western; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D019556:COS Cells; D002478:Cells, Cultured; D004247:DNA / Q000276:immunology*; D019281:Dimerization; D006031:Glycosylation; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D018664:Interleukin-12 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D009154:Mutation / Q000235:genetics*; D009369:Neoplasms / Q000276:immunology / Q000473:pathology; D015202:Protein Engineering; D018417:Th1 Cells / Q000276:immunology; D014162:Transfection
D018664:Interleukin-12; D007371:Interferon-gamma; D004247:DNA