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Novel endonuclease of immune cell, process for producing the same and immune adjuvant using the same

a technology of endonuclease and immune cell, applied in the field of new endonuclease enzyme, can solve the problems of no further studies and knowledge regarding the processing and presentation mechanism of dna antigen, low methylation frequency of cpg dinucleotide present in bacterial dna, and non-production of autoantibody which is cross-reactive to mammalian dsdna

Inactive Publication Date: 2005-09-15
CJ CHEILJEDANG CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, any autoantibody which is cross-reactive to mammalian dsDNA was not produced.
However, there are no further studies and knowledge regarding processing and presentation mechanism of DNA antigen.
Second, methylation frequency of CpG dinucleotide present in bacterial DNA is low.
However, there is no report as to what mechanism enables such critical bacterial DNA to produce antibody and how oligonucleotide having CpG motif is made in cell.

Method used

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  • Novel endonuclease of immune cell, process for producing the same and immune adjuvant using the same
  • Novel endonuclease of immune cell, process for producing the same and immune adjuvant using the same
  • Novel endonuclease of immune cell, process for producing the same and immune adjuvant using the same

Examples

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example 1

Biosynthesis and Secretion of the Endonuclease from Immune Cell

[0060] From the fact that the enzymatic activity of DNase I is widely distributed over human tissue and body fluid, it was assumed that the enzyme possesses certain physiological in vivo functions in addition to the digestive function (Nadano D., et al (1993) Clin. Chem. 39, 448-452; and Yasuda T., et al (1993) Clin. Chim. Acta. 218, 5-16). DNase I is known to cleave internucleosomal DNA during apoptosis (Peitsch M. C., et al (1993) EMBO J. 12, 371-377). The presence of DNase I in human scrum and the biochemical properties thereof were reported (Love J. D., and Hewitt R. R.(1979) J. Biol. Chem. 254, 12588-12594; and Kishi K., et al (1990) Am. J Hum. Genet. 47, 121-126). It was taught that serum DNase I is secreted from pancreas (Love J. D., and Hewitt R. R.(1979) J. Biol. Chem. 254, 12588-12594; and Ito K. et al (1984) J. Biochem. 95, 1399-1406) but studies on the secretion from other tissues are still required. A study...

example 2

Identification of Mg2+-dependent Endonuclease Inducing Internucleosomal DNA Fragmentation

[0074] Apoptosis is defined as specific type of “cell death” such as chromatin condensation, membrane blebbing or chromatin fragmentation as various nucleosome sizes by endonuclease activity (Wyllie, A. H., et al (1984) J. Pathol. 142, 67-77; Wyllie, A. H. (1980) Nature 284, 555-556; and Kerr, J. F. R., et al (I972) Cancer. 26, 239-257).

[0075] Endonuclease activation is significantly responsible for apoptosis process (Arends, M. J., and Wyllie, A, H. (1990) J. Pathol. 136, 593-608). Many researchers suggested that there are various enzymes which involve nucleosome fragmentation. Examples of enzymes involving internucleosomal DNA fragmentation include DNase I (Peitsch M. C., et al (1993) EMBO J. 12, 71-377), DNase II (Torriglia A., (1995) J. Biol. Chem. 270, 28579-28585; and Barry M. A., and Eastman A. (1993) Arch. Biochem. Biophys. 300.440450), and NUC-18 (Wawabata, H., et al (1997) Biochem. B...

example 3

Action of Endonuclease to Foreign DNA in Immune Cell and Characterization of the Reaction Product

[0088] It was known that bacterial DNAs so far recognized as foreign agent include various structure-determining factors which are not present in mammalian DNA and that such factors are involved in the activation of immune cell (Gilkeson, G. S. et al (1995) J. Clin. Invest. 95, 1398-1402; Gilkeson, G. S. et al (1991) Clin. Immunol, Immunopathol. 59, 288-300; Messina, J. P. et al (1993) Cell. Immunol. 147, 148-157; Krieg, A. M. et al (1995) Nature 374,546-549; and Halpern, M. D. et al (1996) Cell. Immunol. 167, 72-79 ). One of the differences of mammalian DNA from bacterial DNA is that the mammalian DNA was subject to considerable CpG restriction and was selectively methylated on cytosine of CpG dinucleotide (Bird, A. P. (1995) Trends Genet. 11, 94-100; Razin, A., and Friedman, J. (1981) Prog. Nucleic Acid Res. Mol. Biol. 25, 33-52; and Han, J. et al (1994) Antisense Res. Dev. 4, 53-65)....

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Abstract

The present invention relates to a novel endonuclease enzyme which is secreted from immune cell and recognizes bacterial DNA as foreign agent and processes it to produce about 10 bp single-stranded oligonucleotide including CpG motif which is involved in immune response. Also, the present invention relates to a process for producing the endonuclease which comprises culturing human B-lymphoblastic IM9 cell line or TPA-treated myelogenous U937 cell line on an appropriate medium to produce the said endonuclease and isolating the said endonuclease from the cell lysate or the culture medium. In addition, the present invention relates to an immune adjuvant comprising about 10 bp single-stranded oligonucleotide having CpG motif produced by treatment of bacterial DNA by the endonuclease.

Description

BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The present invention relates to a novel endonuclease enzyme which is secreted from immune cells and recognizes bacterial DNA as foreign substance and processes it to generate approximately 10 bases single-stranded oligonucleotide including CpG motif known to involve immune response. In addition, the present invention relates to an immune adjuvant comprising the said single-stranded oligonucleotides of approximately 10 bases generated by the said endonuclease enzyme. [0003] 2. Description of the Prior Art [0004] Mammalian animals develop immune systems to defend against foreign agents. The immune systems is classified into natural (nonspecific) immunity or acquired (antigen-specific) immunity. The innate or nonspecific immunity is a primary resistance against diseases caused by one species, and creates defence barrier as four types such as structural, physiological, endocytic and phagocytic, and inflammatory respons...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N9/22
CPCC12N9/22
Inventor JEON, YEONGPARK, WANLEE, NAJUNG, SANGKIM, DOOKWON, HYUNG
Owner CJ CHEILJEDANG CORP
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