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Macrophage derived chemokine (MDC), MDC analogs, MDC inhibitor substances and uses thereof

Inactive Publication Date: 2006-03-16
ICOS CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0040] Moreover, an aspect of the invention includes MDC polypeptide analogs wherein one or more amino acid residues is added, deleted, or replaced from the MDC polypeptides of the present invention, which analogs retain one or more of the biological activities characteristic of the C-C chemokines, especially of MDC. The small size of MDC facilitates chemical synthesis of such polypeptide analogs, which

Problems solved by technology

However, the processes through which chemokines exert their protective effects have not been fully elucidated, and these chemokines in fact may stimulate HIV replication in cells exposed to the chemokines before HIV infection.
The failure rate of drug therapy for Crohn's disease is relatively high, and the disease is often recurrent even in patients receiving surgical intervention.

Method used

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  • Macrophage derived chemokine (MDC), MDC analogs, MDC inhibitor substances and uses thereof
  • Macrophage derived chemokine (MDC), MDC analogs, MDC inhibitor substances and uses thereof
  • Macrophage derived chemokine (MDC), MDC analogs, MDC inhibitor substances and uses thereof

Examples

Experimental program
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example 1

Isolation of a cDNA Encoding MDC

[0119] A partial cDNA for a new C-C chemokine, designated pMP390, was isolated from a macrophage cDNA library as described in U.S. patent application Ser. No. 08 / 939,107, filed Sep. 26, 1997, and in related international publication number WO 96 / 40923, both of which are incorporated herein by reference. Sequence comparisons were performed on Dec. 14, 1994, by the BLAST Network Service of the National Center for Biotechnology Information (e-mail: “blast@ncbi.nlm.nih.gov”), using the alignment algorithm of Altschul et al., J. Mol. Biol., 215: 403-410 (1990). The sequence analysis revealed that a portion of the isolated macrophage cDNA clone designated pMP390 contained a gene sequence having approximately 60-70% identity with previously-identified chemokine genes, including the human MCP-3 gene and rat MIP-1β gene.

[0120] The 2.85 kb cDNA insert of pMP390 was subcloned into the vector pBluescript SK− (Stratagene, La Jolla Calif.) to facilitate complete ...

example 2

Isolation of Additional cDNA Clones Having the Complete MDC Coding Sequence

[0121] Using the pMP390 cDNA clone isolated in Example 1, additional cDNA clones were isolated from the same human macrophage cDNA library, these additional cDNAs containing additional 5′ sequence and encoding the complete amino acid sequence of a macrophage derived chemokine. The additional cloning and sequencing is described in detail in U.S. Ser. No. 08 / 939,107 and WO 96 / 40923, incorporated herein by reference.

[0122] Of the additional clones, clones designated pNT390-12 and pMP390B contained the largest additional 5′ coding sequence, each extending an additional 72 nucleotides upstream of the sequence previously obtained from the cDNA clone pMP390. A composite DNA sequence, herein designated MDC cDNA, was generated by alignment of the pMP390 and pMP390-12 cDNA sequences. This 2923 base pair composite cDNA sequence, and the deduced amino acid sequence of the chemokine MDC, are set forth in SEQ ID NOs: 1 a...

example 3

Determination of MDC Gene Expression in Human Tissues

[0125] Northern blot analysis were conducted to determine the tissues in which the MDC gene is expressed.

[0126] A radiolabeled pMP390 5′ fragment which corresponds to the region of the MDC cDNA encoding the putative mature form of MDC plus 163 bases of the adjacent 3′ noncoding region was used to probe Multiple Tissue Northern blots (Clontech, Palo Alto, Calif.) containing RNA from various normal human tissues. The probe was denatured by boiling prior to use, and the hybridizations were conducted according to the manufacturer's specifications. Autoradiographs were exposed 5 days at −80° C. with 2 intensifying screens.

[0127] The greatest MDC gene expression was observed in the thymus, with much weaker expression detectable in spleen and lung tissues. Expression of MDC in tissue from the small intestine was at even lower levels, and no expression was detected in brain, colon, heart, kidney, liver, ovary, pancreas, placenta, prost...

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Abstract

The present invention provides purified and isolated polynucleotide sequences encoding a novel macrophage-derived C-C chemokine designated “Macrophage Derived Chemokine” (MDC), and polypeptide fragments and analogs thereof. Also provided are materials and methods for the recombinant or synthetic production of the chemokine, fragments, and analogs; and purified and isolated chemokine protein, and polypeptide fragments and analogs thereof. Also provided are antibodies reactive with the chemokine and methods of making and using all of the foregoing. Also provided are assays for identifying modulators of MDC chemokine activity.

Description

[0001] This application is a continuation-in-part of U.S. patent application Ser. No. 09 / 067,447, filed Apr. 28, 1998, and a continuation-in-part of U.S. patent application Ser. No. 08 / 939,107, filed Sep. 26, 1997, (Attorney docket No. 27866 / 34188), and a continuation-in-part of U.S. patent application Ser. No. 08 / 660,542, filed Jun. 7, 1996, and a continuation-in-part of U.S. patent application Ser. No. 08 / 558,658, filed Nov. 16, 1995, and a continuation-in-part of U.S. patent application Ser. No. 08 / 479,620, filed Jun. 7, 1995. These applications are hereby incorporated by reference in their entirety.FIELD OF THE INVENTION [0002] The present invention relates generally to chemokines and more particularly to purified and isolated polynucleotides encoding a novel human C-C chemokine, to purified and isolated chemokine protein encoded by the polynucleotides, to chemokine analogs, to materials and methods for the recombinant production of the novel chemokine protein and analogs, to an...

Claims

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Application Information

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IPC IPC(8): C07H21/04C12P21/02C07K14/52A61K38/19A61K38/00A61K39/00C07K14/715C12N1/21C12N15/19
CPCA01K2217/05A61K38/00A61K39/00C07K2319/02C07K14/7158C07K2319/00C07K14/523A61K2039/505C07K16/24C07K2317/76
Inventor GRAY, PATRICKCHANTRY, DAVIDDEELEY, MICHAELRAPORT, CAROLGODISKA, RONALD
Owner ICOS CORP
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