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Published on: October 30, 2012
Expression cloning of cell surface receptors for the CpG oligodeoxynucleotide/CXCL14 complex
Risa Saito1, Manaka Hasebe2, Takahiko Hara3
1Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan; Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan.
Abstract:
CpG oligodeoxynucleotides (ODNs) induce innate immune responses in dendritic cells and macrophages. A few CpG ODN receptor molecules have been reported. However, recent studies revealed that CXCL14 is required for the function of CpG ODNs in vivo, thereby increasing the importance of cell surface receptors for the CpG ODN/CXCL14 complex. To identify such CpG ODN/CXCL14 receptor molecules, we performed expression cloning using cDNA libraries of mouse dendritic cells and macrophage-derived cells. Consequently, we successfully isolated cDNAs encoding glycoprotein nonmetastatic melanoma protein B (Gpnmb), scavenger receptor class B1 (Scarb1), and immunoglobulin superfamily 8 (Igsf8). Subsequent experiments revealed that Gpnmb only bound to CpG ODNs, while Igsf8, Scarb1, and Cd36 interacted with both CpG ODNs and the CpG ODN/CXCL14 complex. Igsf3 and Rage only bound to the Cy3-ODN2395/CXCL14 complex. These results suggest that intracellular uptake of the CpG ODN/CXCL14 complex is mediated by multiple types of cell surface receptors.
Insights
CpG oligodeoxynucleotides (ODNs) activate immune cells, with CXCL14 enhancing their function. Researchers identified multiple cell surface receptors, including Gpnmb, Scarb1, and Igsf8, that bind CpG ODNs and the CpG ODN/CXCL14 complex for cellular uptake.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- CpG oligodeoxynucleotides (ODNs) are known to stimulate innate immune responses in dendritic cells and macrophages.
- While some CpG ODN receptors are known, the role of CXCL14 in CpG ODN function in vivo highlights the need to identify receptors for the CpG ODN/CXCL14 complex.
Purpose of the Study:
- To identify cell surface receptor molecules involved in the uptake of CpG ODNs and the CpG ODN/CXCL14 complex.
- To elucidate the specific interactions between identified receptors and CpG ODNs or their complexes with CXCL14.
Main Methods:
- Expression cloning using cDNA libraries from mouse dendritic cells and macrophage-derived cells.
- Biochemical assays to determine binding affinities of identified molecules to CpG ODNs and the CpG ODN/CXCL14 complex.
Main Results:
- Glycoprotein nonmetastatic melanoma protein B (Gpnmb), scavenger receptor class B1 (Scarb1), and immunoglobulin superfamily 8 (Igsf8) were identified as cDNAs encoding potential receptors.
- Gpnmb exclusively bound CpG ODNs.
- Igsf8, Scarb1, and Cd36 bound both CpG ODNs and the CpG ODN/CXCL14 complex, while Igsf3 and Rage specifically bound the Cy3-ODN2395/CXCL14 complex.
Conclusions:
- Multiple cell surface receptors mediate the intracellular uptake of the CpG ODN/CXCL14 complex.
- The findings expand our understanding of the molecular mechanisms underlying CpG ODN-mediated immune activation and CXCL14's role.
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