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Membrane cofactor protein (MCP; CD46) expression in transgenic mice
C Kemper1, M Leung, C B Stephensen
1Division of Rheumatology, Washington University School of Medicine, St. Louis 63110-1093, USA.
Abstract:
Human membrane cofactor protein (MCP; CD46) is a widely distributed complement regulator. In the mouse, expression of MCP is largely restricted to the testis while a related, widely expressed protein (Crry) appears to perform MCP's (CD46) regulatory activity. We have developed two mouse strains transgenic for human MCP (CD46) utilizing an approximately 400 kb YAC clone carrying the complete gene. A third mouse strain was generated using an overlapping YAC clone isolated from a second library. The expression of human MCP (CD46) in these mouse strains was characterized by immunohistochemistry, FACS, Western blotting and RT-PCR. No differences were detected in the isoform pattern or distribution among the three strains, although the expression level varied according to how many copies of the gene were integrated. The expression profile closely mimicked that observed in humans, including the same pattern of isoform expression as the donor. In addition, tissue-specific isoform expression in the kidney, salivary gland and brain paralleled that observed in man. The transgenic mice expressed low levels of MCP (CD46) on their E, in contrast to humans but in line with most other primates. These mice should be a useful tool to analyse tissue-specific expression, to establish animal models of infections and to characterize the role of MCP (CD46) in reproduction.
Insights
Transgenic mice expressing human membrane cofactor protein (MCP; CD46) were created. These mice exhibit human-like MCP expression patterns, offering a valuable tool for studying complement regulation and disease models.
Area of Science:
- Immunology
- Genetics
- Molecular Biology
Background:
- Human membrane cofactor protein (MCP; CD46) is a key complement regulator, widely distributed in humans.
- Mouse MCP expression is primarily in the testis, with a related protein (Crry) performing regulatory functions.
- Understanding MCP's role requires models that mimic human expression patterns.
Purpose of the Study:
- To develop mouse models expressing human MCP (CD46).
- To characterize the expression pattern and isoform distribution of human MCP in transgenic mice.
- To assess the utility of these mice for research into MCP function and disease.
Main Methods:
- Generation of transgenic mouse strains using YAC clones containing the human MCP gene.
- Immunohistochemistry, FACS, Western blotting, and RT-PCR to analyze MCP expression.
- Comparison of expression profiles between transgenic mice and human tissues.
Main Results:
- Three strains of transgenic mice expressing human MCP (CD46) were successfully generated.
- Expression levels varied with gene copy number, but isoform patterns and tissue distribution mirrored human expression.
- Tissue-specific isoform expression in kidney, salivary gland, and brain paralleled human patterns.
- Transgenic mice showed low-level expression on erythrocytes, similar to non-human primates.
Conclusions:
- Transgenic mice accurately replicate human MCP (CD46) expression patterns, including tissue-specific isoforms.
- These mice serve as a valuable tool for studying MCP's role in complement regulation, reproduction, and disease.
- The model facilitates research into infections and the function of MCP (CD46) in various physiological processes.
