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AssigneeGenexine, Inc.
InventorsYoung Chul Sung, Sung Hee Lee, Sang Jun Ha, Man Ki Song, Jun Chang
Estimating the term…
The present invention relates to the IL-12p40 subunit mutant gene which can produce IL-12(interleukin 12) of human and mouse origin with high activity, the expression vector including above mutant gene and the use of them to DNA vaccine adjuvant. Particularly, it relates to IL-12p40 mutant gene which inhibits the secretion of IL-12p40 but normally secretes active IL-12p70 by making mutation at Asn-222(human) or Asn-220(mouse) amino acid of IL-12p40, which acts as a competitive inhibitor of active form of IL-12, IL-12p70. Therefore, the IL-12p40 mutant gene of the present invention can be useful for DNA vaccination and gene therapy against various diseases, for example, AIDS, hepatitis C or hepatitis B, cancer, influenza, tuberculosis and malaria, which essentially require cellular immune responses for their therapy.
An isolated nucleicacid comprising a mutated human IL-2p40-encoding sequence, wherein the mutated human IL-12p40-encoding sequence comprises the nucleic acid sequence of SEQ ID NO:
An isolated nucleic acid comprising a mutated mouse IL-12p40-encoding sequence, wherein the mutated mouse IL-12p40-encoding sequence comprises the nucleic acid sequence of SEQ ID NO:
Dependencies are inferred from the claim text.
From the US filing history. Foreign family members aren’t listed.
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof: from viruses
Medicinal preparations containing antigens or antibodies: characterised by the immunostimulating additives, e.g. chemical adjuvants
Antivirals: for RNA viruses
For RNA viruses: for influenza or rhinoviruses
Term ended 15 Mar 2021 (estimated)
This application is a national stage application of PCT/KR01/00408, filed Mar. 15, 2001 under 35 U.S.C. § 371.
The present invention relates to mutant IL-12p40 subunit genes producing highly active human and mouse Interleukin 12(IL-12), expression vector which contains those mutant genes and methods to use thereof for DNA vaccine adjuvant. Particularly, this invention relates to mutant IL-12p40 subunit genes which can secrete immunoactive IL-12p70 normally but reduce secretion of IL-12p40 by inducing mutation for amino acids Asn-222(mouse) or Asn-220(human) of IL-12p40 working as a competitive inhibitor to active IL-12.
IL-12 is secreted by antigen presenting cells (APC) such as macrophages, monocytes and B cells after getting appropriate stimulation, and works as a regulator for many kinds of immune response in vivo. Particularly, IL-12 has a broad range of activities including proliferation of activated type 1 helper T (Th1) and natural killer (NK) cells, regulation of the production of many cytokines, induction of type 1 T helper cell immune responses, differentiation of CD8+ T cells, stimulation of hematopoietic stem cells (Hsieh, C. S. et al., Science, 260:547-549, 1993), and especially, regulation of immune response by promoting lytic activity of cytotoxic T lymphocytes (CTL) and NK cells (Robertson, M. J. and J. Ritz., Oncologist, 1:88-97, 1999; Trinchieri, G., Annu. Rev. Immunol., 13:251-276, 1995). According to recent reports, peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus (HIV)-infected patients produced five-fold less biologically active IL-12 (Chehimi, J. et al., J. Exp. Med., 179:1361-1366, 1994), and mycobacterial and salmonella infections were found to lack of IL-12 receptor expression (de Jong, R. et al., Science, 280:1435-1438). Due to these roles of IL-12, it could induce strong immune response in vivo against virus, bacteria and various cancers. Therefore, the present inventors can expect development of many treatment agents using the present invention.
FIG. 1 shows the sequence homology of amino acids between human and mouse IL-12p40.
FIG. 2 shows p40 and p35 subunits of wild type human IL-12, and a amino acid changes at putative N-glycosylation sites of p40 and p35 subunits on the present invention.
FIG. 3a shows a result of western blot analysis of human IL-12p35 and its derivatives with mutations at putative N-glycosylation sites in cell lysate of the present invention.
To achieve those objects, the present invention provides human or mouse mutant IL-12p40 subunit genes replaced Asn-222(human) or Asn-220(mouse) which is essential for the secretion of IL-12p40 with other amino acids.
The present invention provides gene construct and expression vector containing IRES sequence for co-expression of human or mouse genes of mutated IL-12p40 subunit and subunit itself.
And also, the present invention provides DNA vaccine vector containing and expressing HCV E2 gene, gene construct and expression vector containing p40 subunit gene which has mutated Asn-222(human) or Asn-220(mouse) sites.