In vivo imaging and quantitation of adoptively transferred human antigen-specific T cells transduced to express a human norepinephrine transporter gene Journal Article
| Authors: | Doubrovin, M. M.; Doubrovina, E. S.; Zanzonico, P.; Sadelain, M.; Larson, S. M.; O'Reilly, R. J. |
| Article Title: | In vivo imaging and quantitation of adoptively transferred human antigen-specific T cells transduced to express a human norepinephrine transporter gene |
| Abstract: | Sequential imaging of genetically marked effector cells after adoptive transfer in vivo has greatly enhanced analyses of their biodistribution, growth, and activity both in animal models and in clinical trials of cellular immunotherapy. However, the immunogenicity of cells expressing xenogeneic reporter constructs limits their survival and clinical utility. To address this limitation, we have evaluated a human norepinephrine transporter (hNET) permitting imaging of transduced cells in vivo with a previously approved clinical grade radiolabeled probe, metaiodobenzylguanidine (MIBG). The hNET gene cDNA was cloned from the SK-N-SH cell line and inserted into a bicistronic retroviral vector also encoding green fluorescent protein. Following transfection, human EBV-specific T lymphocytes were fully functional in vitro and also selectively accumulated [123I]MIBG. In nonobese diabetic/severe combined immunodeficient mice bearing human EBV lymphoma xenografts, as few as 104 transduced T cells injected into the tumors could be imaged by single-photon emission computed tomography (SPECT) or positron emission tomography (PET) after i.v. infusion of [123I]MIBG or [124I]MIBG, respectively. When hNET+ EBV-specific T cells were infused i.v., their migration and specific accumulation in EBV + tumors expressing their restricting HLA allele could be imaged by SPECT or PET over 28 days. Image intensity was closely correlated with the number of T cells accumulated in targeted tumors. The use of two reporter probes (MIBG and 2′-deoxy-2′-fluoro-β-D-arabinofuranosyl-5- iodouracil) permitted independent contemporaneous tracking of two distinct EBV-specific T-cell subpopulations expressing different reporter genes (hNET-CD4+ T cells and HSV-TK-CD8+ T cells) in the same animal using three-dimensional nuclear modalities (SPECT and PET). The hNET-based system described may thus have significant potential as a nonimmunogenic reporter for extended repeated quantitative in vivo imaging of transduced cells in man. ©2007 American Association for Cancer Research. |
| Keywords: | controlled study; unclassified drug; human cell; nonhuman; flow cytometry; positron emission tomography; cd8 antigen; cd8+ t lymphocyte; t lymphocyte; t-lymphocytes; mouse; animals; mice; allele; cell survival; cancer immunotherapy; gene expression; green fluorescent protein; animal experiment; animal model; in vivo study; tumor xenograft; mice, scid; cell line, tumor; transfection; imaging system; molecular cloning; cloning, molecular; genetic transfection; antigen specificity; neuroblastoma; reverse transcriptase polymerase chain reaction; quantitative analysis; immunogenicity; nucleotide sequence; cd4+ t lymphocyte; recombinant proteins; cell subpopulation; lymphoma; retrovirus vector; (3 iodobenzyl)guanidine; (3 iodobenzyl)guanidine i 123; (3 iodobenzyl)guanidine i 124; noradrenalin transporter; reporter gene; norepinephrine plasma membrane transport proteins; transplantation, heterologous; adoptive transfer; effector cell; thymidine kinase; cd4 antigen; gene insertion; single photon emission computer tomography; herpes simplex virus; hla antigen; scid mouse; epstein barr virus; complementary dna; tomography, emission-computed, single-photon; 2' deoxy 2' fluoro beta dextro arabinofuranosyl 5 iodoracil; uracil derivative |
| Journal Title: | Cancer Research |
| Volume: | 67 |
| Issue: | 24 |
| ISSN: | 0008-5472 |
| Publisher: | American Association for Cancer Research |
| Date Published: | 2007-12-15 |
| Start Page: | 11959 |
| End Page: | 11969 |
| Language: | English |
| DOI: | 10.1158/0008-5472.can-07-1250 |
| PUBMED: | 18089827 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 18" - "Export Date: 17 November 2011" - "CODEN: CNREA" - "Molecular Sequence Numbers: GENBANK: M65105, NM_001043;" - "Source: Scopus" |
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