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Blood cell production via activation of the hemoglobin scavenger receptor

A technology of erythrocyte lineage and cells, applied in the direction of blood/immune system cells, animal cells, vertebrate cells, etc., can solve the problem of no response to Epo treatment

Inactive Publication Date: 2007-04-25
赫姆索尔有限合伙公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] In addition, many anemias do not respond to Epo therapy

Method used

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  • Blood cell production via activation of the hemoglobin scavenger receptor
  • Blood cell production via activation of the hemoglobin scavenger receptor
  • Blood cell production via activation of the hemoglobin scavenger receptor

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0076] The pharmaceutical compositions may be prepared by methods known per se for the preparation of pharmaceutically acceptable compositions which can be administered to patients, thus combining an effective amount of the active substance in admixture with pharmaceutically acceptable excipients. For example, Remington's Pharmaceutical Sciences describes suitable excipients (Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., USA 1985).

[0077] On this basis, the pharmaceutical composition includes, but is not limited to, the active compound or substance in combination with one or more pharmaceutically acceptable excipients or diluents, and contained at a suitable pH and isotonic with physiological fluids. in the buffer. The pharmaceutical composition also additionally comprises other agents, eg other agents that can stimulate hematopoiesis, erythropoiesis or myelopoiesis, or that can stimulate the growth, proliferation, differentiation and / or mobiliza...

Embodiment 1

[0088] CD34 + Cells express CD163

[0089] CD34 from cryopreserved adult bone marrow (ABM) + Cells were permeabilized with Cytofix / Cytoperm solution (Pharmingen Transduction Laboratories, adivision of BD BioSciences; Becton, Dickson and Company) at 4°C for 30 minutes, washed with fluorescently labeled isotype control antibody (IgG1-FITC) or fluorescently labeled Mac-158 Mouse anti-human CD163 monoclonal antibody (MBS; Mac158-FITC) incubation. In parallel experiments, cells from the same bone marrow samples were also stained extracellularly with fluorescently labeled anti-CD34 (HPCA-2-PE) and anti-CD45 (H130-FITC). Samples were analyzed using an EPICSXL flow cytometer (Beckman Coulter, Inc). The results are shown in Table 1, indicating that more than 95% of myeloid cells are CD34 / CD45 + , most of these cells can also be stained with anti-CD163 antibody.

[0090] Table 1: CD34 + Flow Cytometry Analysis of Bone Marrow Cells

[0091] % CD45 + / CD34 +

Embodiment 2

[0093] Western blot detects CD34 + CD163 expressed by cells

[0094] Resuspend adult bone marrow-derived CD34 in CHAPS buffer containing protease inhibitor cocktail (0.5% CHAPS, 10 mM Tris, 1 mM MgC12, 1 mM EDTA, and 10% glycerol) + cells to prepare cell lysates. Resuspend cells on ice for 30 minutes. Centrifuge the lysate and transfer the supernatant to a new tube. Resuspend the supernatant in non-reducing SDS loading buffer for Western blot analysis. Samples were then run through an 8% polyacrylamide gel and transferred to a nylon membrane. Nylon membranes were blocked with a solution containing nonfat dry milk and then detected with Mac-158 anti-human CD163 antibody followed by goat anti-mouse antibody conjugated to horseradish peroxidase (BIO-RAD Laboratories, Inc.). Bound secondary antibodies were detected using an enhanced chemiluminescence kit from Amersham plc.

[0095] As shown in Figure 1, from CD34 + CD163 immunoreactivity was detected in cell lysates prepare...

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Abstract

The present invention describes methods and compositions for stimulating the growth, proliferation, differentiation and / or mobilization of stem cells and / or progenitor cells. The method involves administering an effective amount of a substance that activates a CD163 hemoglobin scavenger receptor signaling pathway. The methods and compositions are useful for stimulating hematopoiesis and treating a variety of disorders, including cytopenias, anemia, and for preparing cells for transplantation.

Description

[0001] This case is a divisional application, the parent application number is 02801429.4 (international application number is PCT / CA02 / 00411), the application date is March 26, 2002, and the invention title is "production of blood cells by activating hemoglobin scavenger receptors". field of invention [0002] The present invention relates to methods and compositions for stimulating the growth, proliferation, differentiation and / or mobilization of stem cells to produce blood cells. More specifically, the present invention relates to methods of stimulating hematopoiesis comprising stimulating erythropoiesis and myelopoiesis by stimulating stem cells and erythroid and myeloid progenitor cells, respectively. Background of the invention [0003] Hemoglobin scavenger receptors have recently been identified on monocytes and macrophages (Kristiansen, 2001). This receptor clears hemoglobin by mediating endocytosis of the haptoglobin-hemoglobin complex. This receptor, which has bee...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/06A61KA61PC12NC12N5/078C12N5/0789G01N
Inventor S·G·米勒D·贝尔K·E·马修斯
Owner 赫姆索尔有限合伙公司
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