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SPECT/CT Imaging of Mycobacterium tuberculosis Infection with [125I]anti-C3d mAb

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Abstract

Purpose

Diagnosis and therapeutic monitoring of chronic bacterial infection requires methods to detect and localize sites of infection accurately. Complement C3 activation fragments are generated and covalently bound to selective bacterial pathogens during the immune response and can serve as biomarkers of ongoing bacterial infection. We have developed several probes for detecting tissue-bound C3 deposits, including a monoclonal antibody (mAb 3d29) that recognizes the tissue-bound terminal processing fragments iC3b and C3d but does not recognize native circulating C3 or tissue-bound C3b.

Procedures

To determine whether mAb 3d29 could be used to detect chronic Mycobacterium tuberculosis infection non-invasively, aerosol-infected female C3HeB/FeJ mice were injected with [125I]3d29 mAb and either imaged using single-photon emission computed tomography (SPECT)/X-ray computed tomography (CT) imaging at 24 and 48 h after radiotracer injection or being subjected to biodistribution analysis.

Results

Discrete lesions were detected by SPECT/CT imaging in the lungs and spleens of infected mice, consistent with the location of granulomas in the infected animals as detected by CT. Low-level signal was seen in the spleens of uninfected mice and no signal was seen in the lungs of healthy mice. Immunofluorescence microscopy revealed that 3d29 in the lungs of infected mice co-localized with aggregates of macrophages (detected with anti-CD68 antibodies). 3d29 was detected in the cytoplasm of macrophages, consistent with the location of internalized M. tuberculosis. 3d29 was also present within alveolar epithelial cells, indicating that it detected M. tuberculosis phagocytosed by other CD68-positive cells. Healthy controls showed very little retention of fluorescent or radiolabeled antibody across tissues. Radiolabeled 3d29 compared with radiolabeled isotype control showed a 3.5:1 ratio of increased uptake in infected lungs, indicating specific uptake by 3d29.

Conclusion

3d29 can be used to detect and localize areas of infection with M. tuberculosis non-invasively by 24 h after radiotracer injection and with high contrast.

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Acknowledgements

We would like to acknowledge Mariah Klunk for operating the scanner.

Funding

The authors would like to acknowledge funding from the following sources: JHU Musculoskeletal Research Award (CAF), R01 EB020539 (SKJ), P41 EB024495 (MGP), The Stabler Foundation (MGP), and the Alliance for Lupus Research (VMH).

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Authors and Affiliations

  1. The Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins University, 1550 Orleans St. CRB2 493, Baltimore, MD, 21228, USA

    Catherine A. Foss & Martin G. Pomper

  2. Center for Infection and Inflammation Imaging Research, Department of Pediatrics, Johns Hopkins University, Baltimore, MD, 21228, USA

    Catherine A. Foss, Alvaro A. Ordonez, Sanjay K. Jain & Martin G. Pomper

  3. Department of Medicine, University of Colorado Denver, Aurora, CO, USA

    Liudmila Kulik, V. Michael Holers & Joshua M. Thurman

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  1. Catherine A. Foss
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  2. Liudmila Kulik
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Correspondence to Catherine A. Foss.

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Foss, C.A., Kulik, L., Ordonez, A.A. et al. SPECT/CT Imaging of Mycobacterium tuberculosis Infection with [125I]anti-C3d mAb. Mol Imaging Biol 21, 473–481 (2019). https://doi.org/10.1007/s11307-018-1228-5

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