Lipopolysaccharide (LPS), one of the cell wall components of Gram-negative bacteria, is recognized by and interacted with receptors on macrophages. In this paper, we report the trapping of LPS-coated polystyrene particles via optical tweezers and measured its interaction with murine macrophages (J774A.1 cells) for cells pre-treated with extract of Reishi polysaccharides (EORP) vs. those without EORP treatment. Our experimental results indicate that the cellular affinity for LPS increases when the macrophage is pretreated with EORP. We demonstrate for the first time by conventional biological methods and by tracking the dynamics of optically-trapped LPS-coated particles interacting with J774A.1 cells, that EORP not only enhances J774A.1 cells surface expression of TLR4 and CD14, two receptors on macrophages, as well as LPS binding and phagocytosis internalization, but also reduces the adhesion time constant and increases the force constant of the binding interaction. The application of optical tweezers allows us to study the effect on a single cell quantitatively in real-time with a spatial resolution ~1µm within a single cell.

© 2007 Optical Society of America

1. Y. Y. Wang, K. H. Khoo, S. T. Chen, C. C. Lin, C. H. Wong, and C. H. Lin, "Studies on the immunomodulating and antitumor activities of Ganoderma lucidum (Reishi) polysaccharides: functional and proteomic analyses of a fucosecontaining glycoprotein fraction responsible for the activities," Bioorg. Med. Chem. 10, 1057-1062 (2004). [CrossRef]
2. X. F. Bao, X. S. Wang, Q. Dong, J. N. Fang, and X. Y. Li, "Structural features of immunologically active polysaccharides from Ganoderma lucidum," Phytochemistry 59, 175-181 (2002). [CrossRef] [PubMed]
3. H. Y. Hsu, K. F. Hua, C. C. Lin, C. H. Lin, J. Hsu, and C. H. Wong, "Extract of Reishi polysaccharides induces cytokine expression via TLR4-modulated protein kinase signaling pathways," J. Immunol. 173, 5989-5999 (2004). [PubMed]
4. K. Takeda, T. Kaisho1, and S. Akira1, "Toll-like receptors," Annu. Rev. Immunol. 21,335-376 (2003).
5. A. Ashkin, J. Dziedzic, J. Bjorkholm, and S. Chu, "Observation of a single-beam gradient force optical trap for dielectric particles," Opt. Lett. 11, 288-290 (1986). [CrossRef] [PubMed]
6. A. Ashkin, "Acceleration and trapping of particles by radiation pressure," Phys. Rev. Lett. 24, 156-159 (1970). [CrossRef]
7. A. Ashkin, and J. M. Dziedzic, "Optical trapping and manipulation of single cell using infrared laser beams," Nature 330, 769-771 (1987). [CrossRef] [PubMed]
8. A. Ashkin, "History of optical trapping and manipulation of small-neutral particle, atoms, and molecules," IEEE J. Sel. Top. Quantum Electron. 6, 841-856 (2000). [CrossRef]
9. K. C. Neuman, and S. M. Block, "Optical trapping," Rev. Sci. Instrum. 78, 2787-2809 (2004). [CrossRef]
10. H. Kress, E. H. K. Stelzer, G. Griffiths, and A. Rohrbach, "Control of relative radiation pressure in optical traps: Application to phagocytic membrane binding studies," Phy. Rev. E 71, 061927 (2005). [CrossRef]
11. H. Y. Hsu, and M. H. Wen, "Lipopolysaccharide-mediated reactive oxygen species and signal transduction in the regulation of interleukin-1 gene expression," J. Biol. Chem. 277, 22131-22139 (2002). [CrossRef] [PubMed]
12. R. L. Kitchens, and R. S. Munford, "CD14-dependent internalization of bacterial lipopolysaccharide (LPS) is strongly influenced by LPS aggregation but not by cellular responses to LPS," J. Immunol. 160(4), 1920-1928 (1998).
13. E. E. Meyer, M. F. Islam, W. Lau, and H. D. Ou-Yang, "Complexation kinetics of cyclodextrin with hydrophobic molecules confined in an isolated droplet in water," Langmuir 16, 5519-5522 (2000). [CrossRef]
14. L. A. Hough, and H. D. Ou-Yang, "Viscoelasticity of Aqueous Telechelic Poly (ethylene oxide) solutions: relaxation and structure," Phy. Rev. E. 73,031802 (2006). [CrossRef]
15. M. T. Wei, K. F. Hua, J. Hsu, A. Karmenyan, H. Y. Hsu, and A. Chiou, "The interaction of lopopolysaccharide-coated polystyrene particle with membrane receptor proteins on macrophage measured by optical tweezers," Proc. SPIE 6326, 63260T-1-10 (2006).
16. M. T. Wei, T. Y. Tseng, S. L. Liu, A. Karmenyan, and A. Chiou, "From optical tweezers to optical forced oscillation: principles and potential biomedical applications," Proc. SPIE 6447,644707 (2007). [CrossRef]
17. M. T. Wei, and A. Chiou, "Three-dimensional tracking of Brownian motion of a particle trapped in optical tweezers with a pair of orthogonal tracking beams and the determination of the associated optical force constants," Opt. Express 13, 5798-5806 (2005). [CrossRef] [PubMed]

Annu. Rev. Immunol.

• K. Takeda, T. Kaisho1, and S. Akira1, "Toll-like receptors," Annu. Rev. Immunol. 21,335-376 (2003).

Bioorg. Med. Chem.

• Y. Y. Wang, K. H. Khoo, S. T. Chen, C. C. Lin, C. H. Wong, and C. H. Lin, "Studies on the immunomodulating and antitumor activities of Ganoderma lucidum (Reishi) polysaccharides: functional and proteomic analyses of a fucosecontaining glycoprotein fraction responsible for the activities," Bioorg. Med. Chem. 10, 1057-1062 (2004). [CrossRef]

IEEE J. Sel. Top. Quantum Electron.

• A. Ashkin, "History of optical trapping and manipulation of small-neutral particle, atoms, and molecules," IEEE J. Sel. Top. Quantum Electron. 6, 841-856 (2000). [CrossRef]

J. Biol. Chem.

• H. Y. Hsu, and M. H. Wen, "Lipopolysaccharide-mediated reactive oxygen species and signal transduction in the regulation of interleukin-1 gene expression," J. Biol. Chem. 277, 22131-22139 (2002). [CrossRef] [PubMed]

J. Immunol.

• R. L. Kitchens, and R. S. Munford, "CD14-dependent internalization of bacterial lipopolysaccharide (LPS) is strongly influenced by LPS aggregation but not by cellular responses to LPS," J. Immunol. 160(4), 1920-1928 (1998).
• H. Y. Hsu, K. F. Hua, C. C. Lin, C. H. Lin, J. Hsu, and C. H. Wong, "Extract of Reishi polysaccharides induces cytokine expression via TLR4-modulated protein kinase signaling pathways," J. Immunol. 173, 5989-5999 (2004). [PubMed]

Langmuir

• E. E. Meyer, M. F. Islam, W. Lau, and H. D. Ou-Yang, "Complexation kinetics of cyclodextrin with hydrophobic molecules confined in an isolated droplet in water," Langmuir 16, 5519-5522 (2000). [CrossRef]

Nature

• A. Ashkin, and J. M. Dziedzic, "Optical trapping and manipulation of single cell using infrared laser beams," Nature 330, 769-771 (1987). [CrossRef] [PubMed]

Opt. Express

• M. T. Wei, and A. Chiou, "Three-dimensional tracking of Brownian motion of a particle trapped in optical tweezers with a pair of orthogonal tracking beams and the determination of the associated optical force constants," Opt. Express 13, 5798-5806 (2005). [CrossRef] [PubMed]

Opt. Lett.

• A. Ashkin, J. Dziedzic, J. Bjorkholm, and S. Chu, "Observation of a single-beam gradient force optical trap for dielectric particles," Opt. Lett. 11, 288-290 (1986). [CrossRef] [PubMed]

Phy. Rev. E

• H. Kress, E. H. K. Stelzer, G. Griffiths, and A. Rohrbach, "Control of relative radiation pressure in optical traps: Application to phagocytic membrane binding studies," Phy. Rev. E 71, 061927 (2005). [CrossRef]

Phy. Rev. E.

• L. A. Hough, and H. D. Ou-Yang, "Viscoelasticity of Aqueous Telechelic Poly (ethylene oxide) solutions: relaxation and structure," Phy. Rev. E. 73,031802 (2006). [CrossRef]

Phys. Rev. Lett.

• A. Ashkin, "Acceleration and trapping of particles by radiation pressure," Phys. Rev. Lett. 24, 156-159 (1970). [CrossRef]

Phytochemistry

• X. F. Bao, X. S. Wang, Q. Dong, J. N. Fang, and X. Y. Li, "Structural features of immunologically active polysaccharides from Ganoderma lucidum," Phytochemistry 59, 175-181 (2002). [CrossRef] [PubMed]

Proc. SPIE

• M. T. Wei, T. Y. Tseng, S. L. Liu, A. Karmenyan, and A. Chiou, "From optical tweezers to optical forced oscillation: principles and potential biomedical applications," Proc. SPIE 6447,644707 (2007). [CrossRef]

Rev. Sci. Instrum.

• K. C. Neuman, and S. M. Block, "Optical trapping," Rev. Sci. Instrum. 78, 2787-2809 (2004). [CrossRef]

Other

• M. T. Wei, K. F. Hua, J. Hsu, A. Karmenyan, H. Y. Hsu, and A. Chiou, "The interaction of lopopolysaccharide-coated polystyrene particle with membrane receptor proteins on macrophage measured by optical tweezers," Proc. SPIE 6326, 63260T-1-10 (2006).

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