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Enhanced detection of myeloperoxidase activity in deep tissues through luminescent excitation of near-infrared nanoparticles

Nature Medicine volume 19, pages 500–505 (2013)Cite this article

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Abstract

A previous study reported the use of luminol for the detection of myeloperoxidase (MPO) activity using optical imaging in infiltrating neutrophils under inflammatory disease conditions. The detection is based on a photon-emitting reaction between luminol and an MPO metabolite. Because of tissue absorption and scattering, however, luminol-emitted blue light can be efficiently detected from superficial inflammatory foci only. In this study we report a chemiluminescence resonance energy transfer (CRET) methodology in which luminol-generated blue light excites nanoparticles to emit light in the near-infrared spectral range, resulting in remarkable improvement of MPO detectability in vivo. CRET caused a 37-fold increase in luminescence emission over luminol alone in detecting MPO activity in lung tissues after lipopolysaccharide challenge. We demonstrated a dependence of the chemiluminescent signal on MPO activity using MPO-deficient mice. In addition, co-administration of 4-aminobenzoic acid hydrazide (4-ABAH), an irreversible inhibitor of MPO, significantly attenuated luminescent emission from inflamed lungs. Inhibition of nitric oxide synthase with a nonspecific inhibitor, L-NAME, had no effect on luminol-mediated chemiluminescence production. Pretreatment of mice with MLN120B, a selective inhibitor of IKK-2, resulted in suppression of neutrophil infiltration to the lung tissues and reduction of MPO activity. We also demonstrated that CRET can effectively detect MPO activity at deep tissue tumor foci due to tumor development–associated neutrophil infiltration. We developed a sensitive MPO detection methodology that provides a means for visualizing and quantifying oxidative stress in deep tissue. This method is amenable to rapid evaluation of anti-inflammatory agents in animal models.

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Figure 1: Detection of pulmonary inflammation with Luminol-R.
Figure 2: Confirmation that CRET substantially improves the in vivo detectability of MPO activity.
Figure 3: Luminol-R-emitted luminescent signal is dependent on MPO activity.
Figure 4: Effect of MLN120B treatment on MPO activity.
Figure 5: Imaging MPO activity in MDA-MB-231-luc2 tumor metastases.

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Acknowledgements

We thank H. Xu for spectrometer analysis, E. Lim and K. Wong for technical assistance and S. Ray and R. Singh for valuable discussion.

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Author notes

  1. Ning Zhang

    Present address: Present address: In Vivo Pharmacology and Oncology, Pharmaron, Beijing, China.,

Authors and Affiliations

  1. Caliper Life Sciences, Alameda, California, USA

    Ning Zhang, Kevin P Francis & Daniel Ansaldi

  2. Department of Anesthesia and Perioperative Care, San Francisco General Hospital, University of California, San Francisco, California, USA

    Arun Prakash

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Contributions

Experimental design and concepts were devised by N.Z. and K.P.F., D.A. conducted all the experiments except the Mpo−/− study, which was performed by D.A., A.P. and K.P.F., D.A. and N.Z. performed the data analysis. N.Z. wrote the manuscript.

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Correspondence to Ning Zhang.

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The authors declare no competing financial interests.

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Zhang, N., Francis, K., Prakash, A. et al. Enhanced detection of myeloperoxidase activity in deep tissues through luminescent excitation of near-infrared nanoparticles. Nat Med 19, 500–505 (2013). https://doi.org/10.1038/nm.3110

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