Advances in imaging of new targets for pharmacological intervention in stroke: real-time tracking of T-cells in the ischaemic brain

Ortolano, F., Maffia, P. , Dever, G., Rodolico, G. , Millington, O.R., De Simoni, M.G., Brewer, J.M. , Bushell, T.J., Garside, P. and Carswell, H.V. (2010) Advances in imaging of new targets for pharmacological intervention in stroke: real-time tracking of T-cells in the ischaemic brain. British Journal of Pharmacology, 159(4), pp. 808-811. (doi: 10.1111/j.1476-5381.2009.00527.x) (PMID:20015295) (PMCID:PMC2829206)

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Abstract

Background and purpose: T‐cells may play a role in the evolution of ischaemic damage and repair, but the ability to image these cells in the living brain after a stroke has been limited. We aim to extend the technique of real‐time in situ brain imaging of T‐cells, previously shown in models of immunological diseases, to models of experimental stroke. Experimental approach: Male C57BL6 mice (6–8 weeks) (n= 3) received a total of 2–5 × 106 carboxyfluorescein diacetate succinimidyl ester (CFSE)‐labelled lymphocytes from donor C57BL6 mice via i.v. injection by adoptive transfer. Twenty‐four hours later, recipient mice underwent permanent left distal middle cerebral artery occlusion (MCAO) by electrocoagulation or by sham surgery under isoflurane anaesthesia. Female hCD2‐green fluorescent protein (GFP) transgenic mice that exhibit GFP‐labelled T‐cells underwent MCAO. At 24 or 48 h post‐MCAO, a sagittal brain slice (1500 µm thick) containing cortical branches of the occluded middle cerebral artery (MCA) was dissected and used for multiphoton laser scanning microscopy (MPLSM). Key results: Our results provide direct observations for the first time of dynamic T‐cell behaviour in living brain tissue in real time and herein proved the feasibility of MPLSM for ex vivo live imaging of immune response after experimental stroke. Conclusions and Implications: It is hoped that these advances in the imaging of immune cells will provide information that can be harnessed to a therapeutic advantage.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Maffia, Professor Pasquale and Millington, Dr Owain and Rodolico, Dr Gabriella and Garside, Professor Paul and Brewer, Professor James
Authors: Ortolano, F., Maffia, P., Dever, G., Rodolico, G., Millington, O.R., De Simoni, M.G., Brewer, J.M., Bushell, T.J., Garside, P., and Carswell, H.V.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
College of Social Sciences > School of Education
Journal Name:British Journal of Pharmacology
Journal Abbr.:BJP
Publisher:Wiley
ISSN:0007-1188
ISSN (Online):1476-5381
Published Online:10 December 2009

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
404921Investigating the role of T cells in vascular pathologyPaul GarsideBritish Heart Foundation (BHF)PG/6/083/211998Infection Immunity and Inflammation Medicine