Antibody and inhibitor and transformation method and transformation kit using the same

Abstract

The present invention provides a monoclonal antibody which specifically binds to human Toll-like receptor 3. Further, the present invention provides an inhibitor which (a) suppresses a double-stranded RNA-mediated immune response in a cell which expresses Toll-like receptor recognizing the double-stranded RNA and produces type I interferon, and (b) includes an antibody, which binds to the Toll-like receptor and inhibits production of the type I interferon. Particularly, the antibody is a monoclonal antibody against human Toll-like receptor 3. Further, the present invention provides a transfection method comprising the step of infecting a cell which expresses Toll-like receptor recognizing a double-stranded RNA and produces type I interferon by expressing Toll-like receptor recognizing a double-stranded RNA with a recombined virus vector, in which a gene of interest has been inserted, under the inhibitory condition for production of the type I interferon by using an inhibitor including an antibody which binds to the Toll-like receptor. Thus, it is possible to provide (i) an inhibitor which inhibits the production of the type I interferon induced by a virus double-stranded RNA so as to suppress an immune response against the virus, and (ii) a transfection method or a transfection kit whereby a transfection efficiency is improved without enhancing an infectious capacity of a virus vector.

Description

TECHNICAL FIELD [0001] The present invention relates to (i) an antibody specifically bound to human Toll-like receptor 3, (ii) an inhibitor for blocking a signaling induced by double-stranded RNA (ribonucleic acid) so as to suppress a double-stranded RNA-mediated immune response in a cell which produces type I interferon (interferon-α and interferon-β) by expressing the Toll-like receptor recognizing the double-stranded RNA on the surface, (iii) a transfection method or a transfection kit using the antibody and the inhibitor so as to carry out transfection with a recombined RNA virus vector in which a gene of interest is inserted. BACKGROUND ART [0002] It is said that there have been viruses since so early stage of birth of life and have been evolving while contributing to evolution of organisms. There are many viruses having RNA as a gene (RNA viruses), and 90% of plant viruses are RNA viruses. [0003] Recently, a method for introducing genetically-engineered genes into mammalian ce...

AI Technical Summary

Benefits of technology

[0012] The object of the present invention is to provide (i) an antibody against a Toll-like receptor having a function for inhibiting production of type I interferon which is induced by a viral double-stranded RNA, (ii) an inhibitor capable of suppressing immune response against a specific virus by inhibiting the production of the type I interferon induced by the viral double-stranded RNA, and (iii) a transfection method or a transfection kit by which it is possible to improve a transfection efficiency without enhancing an infection efficiency of a virus vector.

[0015] When the antibody or the inhibitor is used, the antibody binds to the Toll-like receptor recognizing the double-stranded RNA so that it is possible to inhibit the binding between the double-stranded RNA and the Toll-like receptor, thereby preventing the type I interferon from being produced in a downstream of signaling pathways involved in the immune response against the double-stranded RNA. Thus, the antibody suppresses the immune response in TLR3-expressing cells induced by the double-stranded RNA.

[0016] Thus, this antibody enables us to amplify RNA viral infection by suppressing the immune response. The suppression of the immune response is not observed in uninfected cells even in case of single-stranded RNA. However, the single-stranded RNA virus has a double-stranded RNA phase during a process of gene replication, so that it is possible to amplify (promote) single-stranded RNA viral infection. Thus, it is feasible to improve a transfection efficiency with a RNA virus vector such as a Sendai-virus vector, a retrovirus vector, and the like without enhancing an infectious efficiency of the virus vector. Note that, examples of the RNA virus include a negative-stranded RNA virus such as Sendai-virus, a positive-stranded RNA virus, and a double-stranded RNA virus. Each of these viruses replicates a large number of double-stranded RNAs in cells, so that infection of these viruses would be amplified by the antibody or the inhibitor.

[0017] Further, the antibody or the inhibitor can bind to the Toll-like receptor recognizing double-stranded RNA thereby suppressing the immune response in upstream of the signaling pathway, which leads to selective suppression of the immune response against the RNA virus. As a result, it is possible to maintain an immune function against antigens other than the RNA virus, e.g., a DNA (deoxyribo nucleic acid) virus, bacteria, and the like.

[0020] According to them, as described above, it is possible to amplify (promote) infection of single-stranded RNA virus and double-stranded RNA virus with the inhibitor. Thus, it is possible to improve a transfection efficiency using RNA virus vector such as retrovirus vector without enhancing an infection efficiency of the virus vector.

Problems solved by technology

However, the improvement of the infection efficiency has not yet been achieved, and is the largest factor which prevents application of the virus vector as a multipurpose vector.

Recently, it is a problem to resolve the mechanism of the immune system how an innate immunity (basic immunity) system of plant, insects, invertebrates, and vertebrates detects and prevents virus invasion.

However, it has not been clarified how the animal cells recognize the viral double-stranded RNA and which signaling pathway produces the type I interferon.

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