Non-invasive tracking of CD4+ T cells with a paramagnetic and fluorescent nanoparticle in brain ischemia

J Cereb Blood Flow Metab. 2016 Aug;36(8):1464-76. doi: 10.1177/0271678X15611137. Epub 2015 Oct 19.


Recent studies have demonstrated that lymphocytes play a key role in ischemic brain injury. However, there is still a lack of viable approaches to non-invasively track infiltrating lymphocytes and reveal their key spatiotemporal events in the inflamed central nervous system (CNS). Here we describe an in vivo imaging approach for sequential monitoring of brain-infiltrating CD4(+) T cells in experimental ischemic stroke. We show that magnetic resonance imaging (MRI) or Xenogen imaging combined with labeling of SPIO-Molday ION Rhodamine-B (MIRB) can be used to monitor the dynamics of CD4(+) T cells in a passive transfer model. MIRB-labeled CD4(+) T cells can be longitudinally visualized in the mouse brain and peripheral organs such as the spleen and liver after cerebral ischemia. Immunostaining of tissue sections showed similar kinetics of MIRB-labeled CD4(+) T cells when compared with in vivo observations. Our results demonstrated the use of MIRB coupled with in vivo imaging as a valid method to track CD4(+) T cells in ischemic brain injury. This approach will facilitate future investigations to identify the dynamics and key spatiotemporal events for brain-infiltrating lymphocytes in CNS inflammatory diseases.

Keywords: CD4+ T cells; In vivo imaging; Ischemic stroke; USPIO; lymphocytes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain Ischemia / blood*
  • Brain Ischemia / diagnostic imaging
  • Brain Ischemia / immunology
  • CD4-Positive T-Lymphocytes / cytology*
  • Cell Tracking / methods*
  • DNA-Binding Proteins / genetics
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • Magnetic Resonance Imaging / methods*
  • Magnetite Nanoparticles / chemistry*
  • Male
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Rhodamines / chemistry
  • Staining and Labeling / methods


  • DNA-Binding Proteins
  • Fluorescent Dyes
  • Magnetite Nanoparticles
  • Rag2 protein, mouse
  • Rhodamines
  • rhodamine B