Feasibility of real-time in vivo 89Zr-DFO-labeled CAR T-cell trafficking using PET imaging

Autoři: Suk Hyun Lee aff001;  Hyunsu Soh aff003;  Jin Hwa Chung aff003;  Eun Hye Cho aff001;  Sang Ju Lee aff001;  Ji-Min Ju aff005;  Joong Hyuk Sheen aff005;  Hyori Kim aff004;  Seung Jun Oh aff001;  Sang-Jin Lee aff005;  Junho Chung aff006;  Kyungho Choi aff006;  Seog-Young Kim aff004;  Jin-Sook Ryu aff001
Působiště autorů: Department of Nuclear Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea aff001;  Department of Radiology, Division of Nuclear Medicine, Hallym University Kangnam Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Republic of Korea aff002;  Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of Korea aff003;  Convergence Medicine Research Center, Asan Medical Center, Seoul, Republic of Korea aff004;  Research Institute, National Cancer Center, Gyeonggi-do, Republic of Korea aff005;  Department of Biomedical Sciences, Seoul National University, Seoul, Republic of Korea aff006
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
prolekare.web.journal.doi_sk: 10.1371/journal.pone.0223814



Chimeric antigen receptor (CAR) T-cells have been recently developed and are producing impressive outcomes in patients with hematologic malignancies. However, there is no standardized method for cell trafficking and in vivo CAR T-cell monitoring. We assessed the feasibility of real-time in vivo 89Zr-p-Isothiocyanatobenzyl-desferrioxamine (Df-Bz-NCS, DFO) labeled CAR T-cell trafficking using positron emission tomography (PET).


The 89Zr-DFO radiolabeling efficiency of Jurkat/CAR and human peripheral blood mononuclear cells (hPBMC)/CAR T-cells was 70%–79%, and cell radiolabeling activity was 98.1–103.6 kBq/106 cells. Cell viability after radiolabeling was >95%. Cell proliferation was not significantly different during the early period after radiolabeling, compared with unlabeled cells; however, the proliferative capacity decreased over time (day 7 after labeling). IL-2 or IFN-γ secretion was not significantly different between unlabeled and labeled CAR T-cells. PET/magnetic resonance imaging in the xenograft model showed that most of the 89Zr-DFO-labeled Jurkat/CAR T-cells were distributed in the lung (24.4% ± 3.4%ID) and liver (22.9% ± 5.6%ID) by one hour after injection. The cells gradually migrated from the lung to the liver and spleen by day 1, and remained stable in these sites until day 7 (on day 7: lung 3.9% ± 0.3%ID, liver 36.4% ± 2.7%ID, spleen 1.4% ± 0.3%ID). No significant accumulation of labeled cells was identified in tumors. A similar pattern was observed in ex vivo biodistributions on day 7 (lung 3.0% ± 1.0%ID, liver 19.8% ± 2.2%ID, spleen 2.3% ± 1.7%ID). 89Zr-DFO-labeled hPBMC/CAR T-cells showed a similar distribution, compared with Jurkat/CAR T-cells, on serial PET images. CAR T cell distribution was cross-confirmed by flow cytometry, Alu polymerase chain reaction, and immunohistochemistry.


Real-time in vivo cell trafficking is feasible using PET imaging of 89Zr-DFO-labeled CAR T-cells. This can be used to investigate cellular kinetics, initial in vivo biodistribution, and safety profiles in future CAR T-cell development.

Klíčová slova:

Basic cancer research – Cancer immunotherapy – Liver – Polymerase chain reaction – Positron emission tomography – Radioactivity – Spleen – T cells


1. Brown CE, Alizadeh D, Starr R, Weng L, Wagner JR, Naranjo A, et al. Regression of glioblastoma after chimeric antigen receptor T-cell therapy. N Engl J Med. 2016; 375: 2561–2569. doi: 10.1056/NEJMoa1610497 28029927

2. Curran KJ, Pegram HJ, Brentjens RJ. Chimeric antigen receptors for T cell immunotherapy: current understanding and future directions. J Gene Med. 2012; 14: 405–415. doi: 10.1002/jgm.2604 22262649

3. Ho WY, Blattman JN, Dossett ML, Yee C, Greenberg PD. Adoptive immunotherapy: engineering T cell responses as biologic weapons for tumor mass destruction. Cancer Cell. 2003; 3: 431–437. doi: 10.1016/s1535-6108(03)00113-2 12781360

4. Sadelain M, Riviere I, Brentjens R. Targeting tumours with genetically enhanced T lymphocytes. Nat Rev Cancer. 2003; 3: 35–45. doi: 10.1038/nrc971 12509765

5. Kershaw MH, Westwood JA, Darcy PK. Gene-engineered T cells for cancer therapy. Nat Rev Cancer. 2013; 13: 525–541. doi: 10.1038/nrc3565 23880905

6. Brentjens RJ, Davila ML, Riviere I, Park J, Wang X, Cowell LG, et al. CD19-targeted T cells rapidly induce molecular remissions in adults with chemotherapy-refractory acute lymphoblastic leukemia. Sci Transl Med. 2013; 5: 177ra138. doi: 10.1126/scitranslmed.3005930 23515080

7. Grupp SA, Kalos M, Barrett D, Aplenc R, Porter DL, Rheingold SR, et al. Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. N Engl J Med. 2013; 368: 1509–1518. doi: 10.1056/NEJMoa1215134 23527958

8. Kalos M, Levine BL, Porter DL, Katz S, Grupp SA, Bagg A, et al. T cells with chimeric antigen receptors have potent antitumor effects and can establish memory in patients with advanced leukemia. Sci Transl Med. 2011; 3: 95ra73. doi: 10.1126/scitranslmed.3002842 21832238

9. De Oliveira SN, Wang J, Ryan C, Morrison SL, Kohn DB, Hollis RP. A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors. J Transl Med. 2013; 11: 23. doi: 10.1186/1479-5876-11-23 23360526

10. Teachey DT, Lacey SF, Shaw PA, Melenhorst JJ, Maude SL, Frey N, et al. Identification of predictive biomarkers for cytokine release syndrome after chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia. Cancer Discov. 2016; 6: 664–679. doi: 10.1158/2159-8290.CD-16-0040 27076371

11. Wang H, Du X, Chen WH, Lou J, Xiao HL, Pan YM, et al. Establishment of a quantitative polymerase chain reaction assay for monitoring chimeric antigen receptor T cells in peripheral blood. Transplant Proc. 2018; 50: 104–109. doi: 10.1016/j.transproceed.2017.11.028 29407291

12. Zheng Z, Chinnasamy N, Morgan RA. Protein L: a novel reagent for the detection of chimeric antigen receptor (CAR) expression by flow cytometry. J Transl Med. 2012; 10: 29. doi: 10.1186/1479-5876-10-29 22330761

13. Adonai N, Adonai N, Nguyen KN, Walsh J, Iyer M, Toyokuni T, et al. Ex vivo cell labeling with 64Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) for imaging cell trafficking in mice with positron-emission tomography. Proc Natl Acad Sci U S A. 2002; 99: 3030–3035. doi: 10.1073/pnas.052709599 11867752

14. Hofmann M, Wollert KC, Meyer GP, Menke A, Arseniev L, Hertenstein B, et al. Monitoring of bone marrow cell homing into the infarcted human myocardium. Circulation. 2005; 111: 2198–2202. doi: 10.1161/01.CIR.0000163546.27639.AA 15851598

15. Abou DS, Ku T, Smith-Jones PM. In vivo biodistribution and accumulation of 89Zr in mice. Nucl Med Biol. 2011; 38: 675–681. doi: 10.1016/j.nucmedbio.2010.12.011 21718943

16. Bansal A, Pandey MK, Demirhan YE, Nesbitt JJ, Crespo-Diaz RJ, Terzic A, et al. Novel 89Zr cell labeling approach for PET-based cell trafficking studies. EJNMMI Res. 2015; 5: 19. doi: 10.1186/s13550-015-0098-y 25918673

17. Roca M, de Vries EF, Jamar F, Israel O, Signore A. Guidelines for the labelling of leucocytes with 111In-oxine: inflammation/infection taskgroup of the European association of nuclear medicine. Eur J Nucl Med Mol Imaging. 2010; 37: 835–841. doi: 10.1007/s00259-010-1393-5 20198474

18. Sato N, Wu H, Asiedu KO, Szajek LP, Griffiths GL, Choyke PL. 89Zr-oxine complex PET cell imaging in monitoring cell-based therapies. Radiology. 2015; 275: 490–500. doi: 10.1148/radiol.15142849 25706654

19. Weist MR, Starr R, Aguilar B, Chea J, Miles JK, Poku E, et al. PET of adoptively transferred chimeric antigen receptor T cells with 89Zr-oxine. Journal of Nuclear Medicine. 2018; 59: 1531–1537. doi: 10.2967/jnumed.117.206714 29728514

20. Parente-Pereira AC, Burnet J, Ellison D, Foster J, Davies DM, van der Stegen S, et al. Trafficking of CAR-engineered human T cells following regional or systemic adoptive transfer in SCID beige mice. J Clin Immunol. 2011; 31: 710–718. doi: 10.1007/s10875-011-9532-8 21505816

21. Fisher B, Packard BS, Read EJ, Carrasquillo JA, Carter CS, Topalian SL, et al. Tumor localization of adoptively transferred indium-111 labeled tumor infiltrating lymphocytes in patients with metastatic melanoma. J Clin Oncol. 1989; 7: 250–261. doi: 10.1200/JCO.1989.7.2.250 2644399

22. Pittet MJ, Grimm J, Berger CR, Tamura T, Wojtkiewicz G, Nahrendorf M, et al. In vivo imaging of T cell delivery to tumors after adoptive transfer therapy. Proc Natl Acad Sci U S A. 2007; 104: 12457–12461. doi: 10.1073/pnas.0704460104 17640914

23. Read EJ, Keenan AM, Carter CS, Yolles PS, Davey RJ. In vivo traffic of indium-111-oxine labeled human lymphocytes collected by automated apheresis. J Nucl Med. 1990; 31: 999–1006. 2112185

24. Smith ME, Ford WL. The recirculating lymphocyte pool of the rat: a systematic description of the migratory behaviour of recirculating lymphocytes. Immunology. 1983; 49: 83–94. 6840811

25. Wagstaff J, Gibson C, Thatcher N, Ford WL, Sharma H, Crowther D. Human lymphocyte traffic assessed by indium-111 oxine labelling: clinical observations. Clin Exp Immunol. 1981; 43: 443–449. 7285388

26. Charoenphun P, Meszaros LK, Chuamsaamarkkee K, Sharif-Paghaleh E, Ballinger JR, Ferris TJ, et al. [89Zr]oxinate4 for long-term in vivo cell tracking by positron emission tomography. Eur J Nucl Med Mol Imaging. 2015; 42: 278–287. doi: 10.1007/s00259-014-2945-x 25359636

27. Hamann A, Klugewitz K, Austrup F, Jablonski-Westrich D. Activation induces rapid and profound alterations in the trafficking of T cells. Eur J Immunol. 2000; 30: 3207–3218. doi: 10.1002/1521-4141(200011)30:11<3207::AID-IMMU3207>3.0.CO;2-L 11093136

28. Looney MR, Thornton EE, Sen D, Lamm WJ, Glenny RW, Krummel MF. Stabilized imaging of immune surveillance in the mouse lung. Nat Methods. 2011; 8: 91–96. doi: 10.1038/nmeth.1543 21151136

29. Morgan RA, Yang JC, Kitano M, Dudley ME, Laurencot CM, Rosenberg SA. Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol Ther. 2010; 18: 843–851. doi: 10.1038/mt.2010.24 20179677

30. Chen Y-P, Zhang Y, Lv J-W, Li Y-Q, Wang Y-Q, He Q-M, et al. Genomic analysis of tumor microenvironment immune types across 14 solid cancer types: immunotherapeutic implications. Theranostics. 2017; 7: 3585–3594. doi: 10.7150/thno.21471 28912897

31. Kircher MF, Gambhir SS, Grimm J. Noninvasive cell-tracking methods. Nat Rev Clin Oncol. 2011; 8: 677–688. doi: 10.1038/nrclinonc.2011.141 21946842

Článok vyšiel v časopise


2020 Číslo 1

Najčítanejšie v tomto čísle

Tejto téme sa ďalej venujú…


Zvýšte si kvalifikáciu online z pohodlia domova

nový kurz
Autori: MUDr. Petr Výborný, CSc., FEBO

Autori: MUDr. Jiří Horažďovský, Ph.D

Zánětlivá bolest zad a axiální spondylartritida – Diagnostika a referenční strategie
Autori: MUDr. Monika Gregová, Ph.D., MUDr. Kristýna Bubová

Krvácení v důsledku portální hypertenze při jaterní cirhóze – od pohledu záchranné služby až po závěrečný hepato-gastroenterologický pohled
Autori: PhDr. Petr Jaššo, MBA, MUDr. Hynek Fiala, Ph.D., prof. MUDr. Radan Brůha, CSc., MUDr. Tomáš Fejfar, Ph.D., MUDr. David Astapenko, Ph.D., prof. MUDr. Vladimír Černý, Ph.D.

Rozšíření možností lokální terapie atopické dermatitidy v ordinaci praktického lékaře či alergologa
Autori: MUDr. Nina Benáková, Ph.D.

Všetky kurzy
Zabudnuté heslo

Nemáte účet?  Registrujte sa

Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.


Nemáte účet?  Registrujte sa