Tat-functionalized near-infrared emissive polymersomes for dendritic cell labeling.
Dendritic cells (DCs) play a pivotal role in both immune tolerance and the initiation of immunological responses. The ability to track DCs in vivo is imperative for the development of DC-based cellular therapies and to advance our understanding of DC function and pathophysiology. Here, we conjugate a cell permeable peptide, Tat, to near-infrared (NIR) emissive polymersomes in order to enable efficient intracellular delivery for future DC tracking with these optical probes. NIR imaging allows quantitative, repetitive, in vivo detection of fluorophore-laden cells, at centimeter tissue depths without disturbing cellular function. Flow cytometry and confocal microscopy results indicate that Tat-mediated polymersome delivery to DCs is concentration and time dependent, resulting in punctate intracellular localization. Further, loading cells with Tat NIR emissive polymersomes does not interfere with cytokine-induced DC maturation and has modest effects on DC viability, but has a significant effect on mature DC-induced activation of naive T cells. We observe significant uptake of NIR emissive polymersomes when conjugated to the peptide, with a lower detection limit of 5000 labeled DCs. The extent of polymersome delivery is estimated as 70 000 +/- 10 000 vesicles/cell, equivalent to 0.7 +/- 0.1 fmol of NIR fluorophore. Our studies will enable future in vivo tracking of ex vivo labeled DCs by NIR fluorescence based imaging.
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Related Subject Headings
- Time Factors
- Spectroscopy, Near-Infrared
- Phenotype
- Organic Chemistry
- Molecular Structure
- Microscopy, Electron, Transmission
- Lymphocyte Culture Test, Mixed
- Humans
- Gene Products, tat
- Dendritic Cells
Citation
Published In
DOI
EISSN
ISSN
Publication Date
Volume
Issue
Start / End Page
Related Subject Headings
- Time Factors
- Spectroscopy, Near-Infrared
- Phenotype
- Organic Chemistry
- Molecular Structure
- Microscopy, Electron, Transmission
- Lymphocyte Culture Test, Mixed
- Humans
- Gene Products, tat
- Dendritic Cells