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Am J Physiol Cell Physiol 293: C1698-C1708, 2007. First published September 26, 2007; doi:10.1152/ajpcell.00215.2007
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VASCULAR BIOLOGY

In vivo leukocyte labeling with intravenous ferumoxides/protamine sulfate complex and in vitro characterization for cellular magnetic resonance imaging

Y. Jeffrey Wu,2 Leslie L. Muldoon,2,3 Csanad Varallyay,2 Sheila Markwardt,1 Richard E. Jones,1,2 and Edward A. Neuwelt1,2,4

1Research Service, Veterans Administration Medical Center, and Departments of 2Neurology, 3Cell and Developmental Biology, and 4Neurosurgery, Oregon Health and Sciences University, Portland, Oregon

Submitted 24 May 2007 ; accepted in final form 21 September 2007

Cellular labeling with ferumoxides (Feridex IV) superparamagnetic iron oxide nanoparticles can be used to monitor cells in vivo by MRI. The objective of this study was to use histology and MRI to evaluate an in vivo, as opposed to in vitro, technique for labeling of mononuclear leukocytes as a means of tracking inflammatory processes in the brain. Long-Evans rats were intravenously injected with 20 mg/kg ferumoxides, ferumoxtran-10, or ferumoxytol with or without protamine sulfate. Leukocytes and splenocytes were evaluated by cell sorting and iron histochemistry or were implanted into the brain for MRI. Injection of ferumoxides/protamine sulfate complex IV resulted in iron labeling of leukocytes (ranging from 7.4 ± 0.5% to 12.5 ± 0.9% with average 9.2 ± 0.8%) compared with ferumoxides (ranging from 3.9 ± 0.4% to 6.3 ± 0.5% with average 5.0 ± 0.5%) or protamine sulfate alone (ranging from 0% to 0.9 ± 0.7% with average 0.3 ± 0.3%). Cell sorting analysis indicated that iron-labeled cells were enriched for cell types positive for the myelomonocytic marker (CD11b/c) and the B lymphocyte marker (CD45RA) and depleted in the T cell marker (CD3). Neither ferumoxtran-10 nor ferumoxytol with protamine sulfate labeled leukocytes. In vivo ferumoxides/protamine sulfate-loaded leukocytes and splenocytes were detected by MRI after intracerebral injection. Ferumoxides/protamine complex labeled CD45RA-positive and CD11b/c-positive leukocytes in vivo without immediate toxicity. The dose of feumoxides in this report is much higher than the approved human dose, so additional animal studies are required before this approach could be translated to the clinic. These results might provide useful information for monitoring leukocyte trafficking into the brain.

leukocyte trafficking; brain; B lymphocyte; rat


Address for reprint requests and other correspondence: E. A. Neuwelt, Dept. of Neurology and Blood-Brain Barrier Program, Oregon Health and Sciences Univ., 3181 Sam Jackson Pkwy., L603, Portland, OR 97239 (e-mail: neuwelte{at}ohsu.edu)






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