In vivo monitoring the fate of Cy5.5-Tat labeled T lymphocytes by quantitative near-infrared fluorescence imaging during acute brain inflammation in a rat model of experimental autoimmune encephalomyelitis.
T cells and macrophages directed against myelin proteins orchestrate the inflammation process in multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). So far, assessment of macrophages infiltration or structural alterations has been achieved by in vivo imaging. In this work, we show the infiltration of Cy5.5-labeled T lymphocytes into the brains of EAE rats by reflectance near-infrared fluorescence imaging. T lymphocytes were labeled with Cy5.5-Tat and administered intravenously to naïve or EAE animals. The highest fluorescence signal was observed for EAE animals, which received myelin-activated T cells during the acute phase of the disease. The temporal profile of fluorescence in this group paralleled the pattern of neurological impairment during the acute phase, the remittance and first relapses of EAE. No disease specific fluorescence pattern was observed for EAE animals, which received naïve T cells. However, uptake of Cy5.5-Tat by scavenger cells (e.g. macrophages) following death of labeled T cells in vivo prevents prolonged longitudinal studies. Our work demonstrates that Cy5.5-Tat labeling of T cells is suitable for in vivo fluorescence imaging of inflammation initiation in the EAE model. This approach may particularly be useful for evaluation of novel anti-inflammatory therapies.
Berger
Cedric
C
Novartis Institutes for Biomedical Research, Basel, Switzerland.
Gremlich
Hans-Ulrich
HU
Schmidt
Philipp
P
Cannet
Catherine
C
Kneuer
Rainer
R
Hiestand
Peter
P
Rausch
Martin
M
Rudin
Markus
M
D000818:Animals; D002232:Carbocyanines; D004660:Encephalitis / Q000276:immunology / Q000473:pathology*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000276:immunology / Q000473:pathology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique / Q000379:methods; D015696:Gene Products, tat; D006678:HIV / Q000235:genetics; D007158:Immunologic Techniques; D018613:Microscopy, Confocal; D051381:Rats; D011917:Rats, Inbred Lew; D019265:Spectroscopy, Near-Infrared; D013601:T-Lymphocytes / Q000276:immunology*; D054322:tat Gene Products, Human Immunodeficiency Virus
C098793:CY5.5 cyanine dye; D002232:Carbocyanines; D015696:Gene Products, tat; D054322:tat Gene Products, Human Immunodeficiency Virus