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Transgenic mice expressing human cd20 and/or cd16

A transgenic animal and genome technology, applied in genetic engineering, plant genetic improvement, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve the problem of lack of co-expressed human marker animal model and so on

Inactive Publication Date: 2006-03-15
GENENTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Despite the prominent roles of CD20 and CD16 in human lymphoma and induction of important immune responses, animal models co-expressing the human markers are lacking

Method used

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  • Transgenic mice expressing human cd20 and/or cd16
  • Transgenic mice expressing human cd20 and/or cd16
  • Transgenic mice expressing human cd20 and/or cd16

Examples

Experimental program
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Effect test

Embodiment approach

[0070] The present invention provides transgenic animals expressing heterologous CD20 markers, heterologous CD16 markers or both. In one embodiment, the human CD20 transgenic animal of the present invention expresses CD20 on the same type of B cells. Administration of anti-human CD20 antibodies to this currently described transgenic animal resulted in the depletion of lymphocytes expressing human CD20B. Due to the inability to incorporate effective transcription control regions into the transgene, previous attempts to prepare transgenic mice expressing human CD20 failed to achieve the expression of human CD20 in a suitable B cell subpopulation. In another embodiment, the transgenic mouse of the invention expresses human CD20 and human CD16.

[0071] The present invention provides transgenic animals that have cells that respond in a manner similar to human subjects. Administration of anti-human CD20 antibodies to this currently described transgenic animal resulted in the depletion ...

Embodiment 1

[0157] This example describes the generation of human CD20BAC transgenic (Tg+) mice, and studies the effect of anti-human CD20 antibody treatment in hCD20+ mice.

[0158] Human CD20 CITB human BAC-D-clone No. 117H19 from Invitrogen (Invitrogen, Carlsbad, CA) was used to produce human CD20 transgenic mice. DNA encoding human CD20 was isolated from human lymphocytes and sent to Invitrogen. Invitrogen tested the DNA with filter paper with clones from the human BAC library and identified clone 117H19. Previous attempts to produce transgenic mice expressing human CD20 were unsuccessful, perhaps partly due to the inability to incorporate sufficient transcription control regions into the transgenic construct. Transgenic mice were produced by microinjecting the human CD20BAC construct prepared by clone 117H19 into the fertilized eggs of the mouse FVB inbred line. The fertilized eggs are incubated for 1-7 days, and then transplanted into surrogate mice. Mice were screened based on FACS ana...

Embodiment 2

[0171] This example shows the synergy between anti-CD20 monoclonal antibody and BR3 antagonist treatment on B cell modulation / depletion. BR3-Fc is an immunoadhesin in which the extracellular domain of human BR3 is fused to the constant domain of an immunoglobulin sequence (in this case, human IgG1).

[0172] Transgenic mice expressing human CD20 (named hCD20 + Mouse) with anti-CD20 monoclonal antibody (100 μg single injection on day 9), BR3-Fc (from day 1 to day 12, 100 μg every other day), or anti-CD20 monoclonal antibody and BR3- The combination of Fc is treated by intraperitoneal injection. Each group consists of 4 mice. Two days after the last injection, kill the mice and analyze hCD20 + B cells. For B cell markers (CD21 + CD23 + ) FACS analysis of spleen, blood, lymph nodes and Peyer’s spots.

[0173] The results showed that anti-CD20 monoclonal antibody treatment reduced more than 99% of mature circulating B cells in the blood and lymph nodes, and BR3-Fc treatment reduced ma...

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Abstract

The present invention relates to immunoglobulin anamorphosis and uses thereof. The invenion provides a humanied and embeded aiti-CD20 body used for treating positive malignant tumor and auto-immune disease.

Description

[0001] This application has been filed as a PCT international patent application on December 11, 2003 in the following names: Gentech Corporation, which is a U.S. national company and resident (applicants in all countries except the United States); Andrew C-Y. Chen , a U.S. citizen and resident (for U.S. applicants only); Gong Qian, a Chinese citizen and U.S. resident (for U.S. applicants only); and Flavius ​​Martin, a Romanian citizen and U.S. resident (for U.S. applicants only) Applicant); this application designates all countries and claims the following priority: U.S. Provisional Application Serial No. 60 / 434,115, filed on December 16, 2002; and U.S. Provisional Application Serial No. 60 / 476,481, filed on June 5, 2003 Submitted on the day. Background technique [0002] Both T and B cells contain cell surface proteins that can be used as differentiation and recognition markers. One such human B cell marker is the human B lymphocyte restricted differentiation antigen Bp35, ...

Claims

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Application Information

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IPC IPC(8): G01N33/00C12P21/00A01K67/033A01K67/027C12N15/00A61KA61K39/395C07H21/04C07K16/00C07K16/28C12N15/63
CPCC07K2316/52C07K2317/565C07K2317/52C07K2317/92C07K2316/95A61K2039/505C07K16/2887C07K2317/72C07K2317/41C07K2317/55C07K2317/56C07K2317/24
Inventor 安德鲁·C-Y·陈龚谦弗莱维厄斯·马丁
Owner GENENTECH INC
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