Normal T-cell response and in vivo magnetic resonance imaging of T cells loaded with HIV transactivator-peptide-derived superparamagnetic nanoparticles

The present study analyzed the feasibility of using magnetic resonance imag ing (MRI) to monitor T-cell homing in vivo after loading T cells with super paramagnetic iron oxide (CLIO) nanoparticles derivatized with a peptide seq uence from the transactivator protein (Tat) of HIV-1. T cells were isolated from C57BL/6 (B6) mice and loaded with 0, 400, 800, 1600, or 8000 ng/ml of FITC conjugated CLIO-Tat (FITC-CLIO-Tat). There was a dose-dependent uptak e of FITC-CLIO-Tat by T cells. Stimulation of FITC-CLIO-Tat loaded T cells with anti-CD3 (0.1 mug/ml) plus IL-2 (5 ng/ml) elicited normal activation a nd activation-induced cell death (AICD) responses, and normal upregulation of CD69, ICAM-1 (CD54), L-selectin (CD62L), and Fas. The FITC-CLIO-Tat load ed T cells (3 X 10(7)) were transferred intravenously (i.v.) into B6 mice a nd the in vivo MRI of mice was acquired using a spin-echo pulse sequence at 4.7 T with a Bruker Biospec system. Homing of T cells into the spleen was observed by a decrease in MRI signal intensity within I It after the transf er, which remained decreased for 2-24 h after transfer. These homing data w ere confirmed by FACS analysis and biodistribution analysis using I-125-CLI O-Tat. Thus, T cells can be efficiently loaded with FITC-CLIO-Tat without i nterfering with their normal activation and AICD, or homing to the spleen, and the biodistribution of FITC-CLIO-Tat loaded T cells can be monitored in vivo over time by MRI. (C) 2001 Elsevier Science B.V. All rights reserved.