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Remedy for Chronic Inflammation and Antibody to be Used Therein

a chronic inflammation and antibody technology, applied in the field of chronic inflammation drugs, can solve the problems of inconsistent effects of conventionally-known anti-tenascin-c antibodies against inflammation, no details given, and no clear finding regarding their relationship with chronic inflammation

Inactive Publication Date: 2011-12-01
TOKYO UNIVERSITY OF SCIENCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013]The present invention can provide the novel remedy against chronic inflammation.

Problems solved by technology

However, no details have been given as to the roles of tenascin-C and the peptide derived therefrom in inflammatory sites, and in particular, there has been obtained no clear finding regarding their relationship with chronic inflammation.
Further, there are cases that the effects of conventionally-known anti-tenascin-C antibodies against inflammation are inconsistent depending on the site.

Method used

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  • Remedy for Chronic Inflammation and Antibody to be Used Therein

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0045]A peptide having the amino acid sequence of SEQ ID NO: 3 was synthesized by a usual method. The synthesized peptide as hapten was fused with KLH, and the obtained fusion peptide was used as an antigenic peptide. Immunization was conducted in accordance with a usual method using rabbits. The mixture including a target antibody obtained was purified by a usual method.

[0046]In the manner as above, three kinds (No. 1 to No. 3) of polyclonal antibodies (anti-human TNIIIA2 antibodies) against the human TNIIIA2 were obtained.

[0047]

[0048]Antibody activity of the anti-human TNIIIA2 polyclonal antibodies obtained above were evaluated as follows by using, as indicators, their inhibition effects against cell adhesion induced by the human TNIIIA2. The results are shown in Table 1.

[0049]Using RPMI 1640 (20% FBS added) as a cell culture medium, KOP 2.16 cells (a mouse bone marrow-derived vascular endothelial cell line) were seeded in each well of a 96-hole plate at a density of 5×104 cells / 2...

example 3

[0069]The effect of the anti-human TNIIIA2 antibody on changes in viable macrophage cell counts over time in the presence of human tenascin-C was evaluated as follows.

[0070]THP-1 cells were cultured in a complete medium, to which PMA was added at a concentration of 10 ng / mL, at 37° C. and 5% CO2 for 24 hours to be induced to differentiate into macrophages (hereinafter, the differentiated THP-1 is referred to as “PMA-THP-1”).

[0071]Into each well of a 96-hole plate, fibronectin (FN) was added at a concentration of 5 μg / mL, and the plate was incubated at 37° C. for 1 hour. The plate was blocked by adding BSA at a concentration of 2.5 mg / mL and incubating at 37° C. for 1 hour, and washed three times with PBS to obtain a plate with FN-coated each well.

[0072]Into the FN-coated wells, an RPMI 1640 (serum-free) culture medium, to which human tenascin-C was added such that its final concentration was 2 μg / mL, was added, and a normal rabbit IgG, the 4F10TT antibody or the anti-human TNIIIA2 a...

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Abstract

The invention provides a remedy for chronic inflammation and an anti-TNIIIA2 antibody to be used therein. The remedy includes an antibody recognizing TNIIIA2, that is a peptide derived from a partial sequence A2 of a human tenascin-C fibronectin III-like repetitive sequence and having the amino acid sequence RSTDLPGLKAATHYTITIRGVC (SEQ ID NO: 1).

Description

TECHNICAL FIELD[0001]The present invention relates to a remedy for chronic inflammation and a novel antibody to be used therein.BACKGROUND ART[0002]The expression of tenascin-C is rarely observed in healthy cells, except for the immune system. Expression of tenascin-C is induced under pathological conditions such as inflammation and tumor growth.[0003]As a method for detecting tenascin-C, some anti-tenascin-C antibodies have been known (for example, see JP-A Nos. 2004-217546 and 2002-234900). An arthritis diagnosis method using tenascin-C as a marker has been also known (for example, see JP-A-2004-138489).[0004]A polypeptide which forms tenascin-C includes an epidermal growth factor-like domain, a fibronectin (FN) III-like domain and a fibrinogen-like domain. Among them, it has been known that the FN III-like domain includes a continuation between 8 basic types of repetitive sequences (1 to 8) and 9 types of repetitive sequences to be spliced (A1, A2, A3, A4, B, AD2, AD1, C and D), ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/18
CPCC07K16/18C07K2317/76C12N5/0634A61P29/00A61P43/00
Inventor FUKAI, FUMIO
Owner TOKYO UNIVERSITY OF SCIENCE
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