Purpose:To develop a clinically translatable method of cell labeling with zirconium 89 ( 89 Zr) and oxine to track cells with positron emission tomography (PET) in mouse models of cell-based therapy. Materials andMethods:This study was approved by the institutional animal care committee. 89Zr-oxine complex was synthesized in an aqueous solution. Cell labeling conditions were optimized by using EL4 mouse lymphoma cells, and labeling efficiency was examined by using dendritic cells (DCs) (n = 4), naïve (n = 3) and activated (n = 3) cytotoxic T cells (CTLs), and natural killer (NK) (n = 4), bone marrow (n = 4), and EL4 (n = 4) cells. The effect of ] cells, n = 4) and by measuring the tumor size (n = 6). Two-way analysis of variance was used to compare labeling conditions, the Wilcoxon test was used to assess cell survival and proliferation, and Holm-Sidak multiple tests were used to assess tumor growth and perform biodistribution analyses. Results:89 Zr-oxine complex was synthesized at a mean yield of 97.3% 6 2.8 (standard deviation). It readily labeled cells at room temperature or 4°C in phosphate-buffered saline (labeling efficiency range, 13.0%-43.9%) and was stably retained (83.5% 6 1.8 retention on day 5 in DCs). Labeling did not affect the viability of DCs and CTLs when compared with nonlabeled control mice (P . .05), nor did it affect functionality. 89Zr-oxine complex enabled extended cell tracking for 7 days. Labeled tumor-specific CTLs accumulated in the tumor (4.6% on day 7) and induced tumor regression (P , .05 on day 7). Conclusion:We have developed a 89Zr-oxine complex cell tracking technique for use with PET that is applicable to a broad range of cell types and could be a valuable tool with which to evaluate various cell-based therapies.q RSNA, 2015
Purpose The success of hematopoietic stem cell transplantation (HSCT) depends on donor cell homing to the bone marrow (BM). However, there is no reliable method of noninvasively monitoring the kinetics and distribution of transferred cells. Using Zirconium-89 (89Zr)-oxine cell labeling combined with positron emission tomography (PET) imaging, we sought to visualize and quantify donor cell homing in a mouse BM transplantation model. Experimental Design The effect of 89Zr-oxine labeling on BM cell viability and differentiation was evaluated in vitro. 89Zr-labeled BM cells (2×107 cells, 16.6 kBq/106 cells) were transferred intravenously and serial microPET images were obtained (n=5). The effect of a CXCR4 inhibitor, plerixafor, (5 mg/kg) and granulocyte-colony stimulation factor (G-CSF, 2.5 μg) on BM homing and mobilization were examined (n=4). Engraftment of the transferred 89Zr-labeled cells was evaluated (n=3). Results 89Zr-oxine-labeled BM cells showed delayed proliferation, but differentiated normally. Transferred BM cells rapidly migrated to the BM, spleen, and liver (n=5). Approximately 36% of donor cells homed to the BM within 4 h, irrespective of prior BM ablation. Inhibition of CXCR4 by plerixafor alone or with G-CSF significantly blocked the BM homing (p<0.0001, vs non-treated, at 2 h), confirming a crucial role of the CXCR4-CXCL12 system. Mobilization of approximately 0.64% of pre-transplanted BM cells induced a 3.8-fold increase of circulating BM cells. 89Zr-labeled donor cells engrafted as well as non-labeled cells. Conclusions 89Zr-oxine PET imaging reveals rapid BM homing of transferred BM cells without impairment of their stem cell functions, and thus, could provide useful information for optimizing HSCT.
BackgroundBone fracture healing is dependent upon the rapid migration and engraftment of bone marrow (BM) progenitor and stem cells to the site of injury. Stromal cell-derived factor-1 plays a crucial role in recruiting BM cells expressing its receptor CXCR4. Recently, a CXCR4 antagonist, plerixafor, has been used to mobilize BM cells into the blood in efforts to enhance cell migration to sites of injury presumably improving healing. In this study, we employed zirconium-89 (89Zr)-oxine-labeled BM cells imaged with positron emission tomography (PET)/computed tomography (CT) to visualize and quantitate BM cell trafficking following acute bone injury and to investigate the effect of plerixafor on BM cell homing. Unilateral 1-mm incisions were created in the distal tibia of mice either on the same day (d0) or 24 h (d1) after 89Zr-oxine-labeled BM cell transfer (n = 4–6, 2–2.3 × 107 cells at 9.65–15.7 kBq/106 cells). Serial microPET/CT imaging was performed and migration of 89Zr-labeled cells to the bone injury was quantified. The effects of three daily doses of plerixafor on cell trafficking were evaluated beginning on the day of fracture generation (n = 4–6). The labeled cells localizing to the fracture were analyzed by flow cytometry and immunohistochemistry.ResultsIn d0- and d1-fracture groups, 0.7% and 1.7% of administered BM cells accumulated within the fracture, respectively. Plerixafor treatment reduced BM cell migration to the fracture by approximately one-third (p < 0.05 for both fracture groups). Flow cytometry analysis of donor cells collected from the injured site revealed a predominance of CD45+ stem/progenitor cell populations and subsequent histological analysis demonstrated the presence of donor cells engrafted within sites of fracture repair.Conclusion89Zr-oxine labeling enabled visualization and quantitation of BM cell recruitment to acute fractures and further demonstrated that plerixafor plays an inhibitory role in this recruitment.Electronic supplementary materialThe online version of this article (10.1186/s13550-018-0463-8) contains supplementary material, which is available to authorized users.
<p>Supplemental Materials and Methods, Supplemental Figures.</p>
<p>Supplemental Materials and Methods, Supplemental Figures.</p>
Systemic immune activation and memory cell generation is desired in dendritic cell (DC)-based therapies against cancers, especially with metastasis. Migration properties of DCs, however, could limit the responses regionally constrained. Clinically, DC-vaccines are often given subcutaneously (s.c.), intravenously (i.v.) or intradermally (i.d.), without monitoring the migration of DCs. In this study, we visualized trafficking of DCs labeled with In-111 oxine using a whole-body micro-SPECT imager after immunization via different routes and examined induction of immune activation and memory cell generation in mice. Bone marrow-derived DCs were used. Surprisingly, most of s.c. transferred DCs failed to migrate to the adjacent lymph nodes (LNs) and died at the injection site, irrespective of their maturation status. DCs injected i.v. were initially trapped in the lungs, but gradually migrated to the spleen, liver and, to a lesser extent, LNs. In contrast, DCs administered i.d. and intraperitoneally (i.p.) quickly entered lymphatic system and distributed to LNs. Proliferation of CMFDA labeled OT-1 CD8 T cells pre-transferred to mice receiving ovalbumin loaded DCs (OVA-DCs) indicated that i.v. and i.p. immunizations triggered stronger and more systemic expansion of effector T cells than s.c. and i.d. immunizations. To examine the establishment of long-term defense, we next immunized mice with OVA-DCs and challenged them with OVA-expressing tumors intramuscularly, contra-lateral to the s.c. and i.d. injection site. Intriguingly, i.v. immunization seemed less effective than the others in developing long-term antigen-specific immunity, despite its capability to induce an extensive acute responses. These results might suggest that difference in the microenvironment of organs where CD8 T cells encounter the antigen (i.e., spleen or LNs) controls the generation of long-lived memory cells. Collectively, immunization route would be a critical factor that determines the spatial distribution of immune activation and generation of functional memory cells in DC-based therapies. Citation Format: Noriko Sato, Kingsley Asiedu, Peter L. Choyke. Differential trafficking of dendritic cells after immunization determines establishment of systemic anti-tumor responses. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2541. doi:10.1158/1538-7445.AM2014-2541
<div>Abstract<p><b>Purpose:</b> The success of hematopoietic stem cell transplantation (HSCT) depends on donor cell homing to the bone marrow. However, there is no reliable method of noninvasively monitoring the kinetics and distribution of transferred cells. Using zirconium-89 (<sup>89</sup>Zr)-oxine cell labeling combined with PET imaging, we sought to visualize and quantify donor cell homing in a mouse bone marrow transplantation model.</p><p><b>Experimental Design:</b> The effect of <sup>89</sup>Zr-oxine labeling on bone marrow cell viability and differentiation was evaluated <i>in vitro</i>. <sup>89</sup>Zr-labeled bone marrow cells (2 × 10<sup>7</sup> cells, 16.6 kBq/10<sup>6</sup> cells) were transferred intravenously, and serial microPET images were obtained (<i>n</i> = 5). The effect of a CXCR4 inhibitor, plerixafor (5 mg/kg) and G-CSF (2.5 μg) on bone marrow homing and mobilization were examined (<i>n</i> = 4). Engraftment of the transferred <sup>89</sup>Zr-labeled cells was evaluated (<i>n</i> = 3).</p><p><b>Results:</b> <sup>89</sup>Zr-oxine–labeled bone marrow cells showed delayed proliferation, but differentiated normally. Transferred bone marrow cells rapidly migrated to the bone marrow, spleen, and liver (<i>n</i> = 5). Approximately 36% of donor cells homed to the bone marrow within 4 hours, irrespective of prior bone marrow ablation. Inhibition of CXCR4 by plerixafor alone or with G-CSF significantly blocked the bone marrow homing (<i>P</i> < 0.0001, vs. nontreated, at 2 hours), confirming a crucial role of the CXCR4–CXCL12 system. Mobilization of approximately 0.64% of pretransplanted bone marrow cells induced a 3.8-fold increase of circulating bone marrow cells. <sup>89</sup>Zr-labeled donor cells engrafted as well as nonlabeled cells.</p><p><b>Conclusions:</b> <sup>89</sup>Zr-oxine PET imaging reveals rapid bone marrow homing of transferred bone marrow cells without impairment of their stem cell functions, and thus, could provide useful information for optimizing HSCT. <i>Clin Cancer Res; 23(11); 2759–68. ©2016 AACR</i>.</p></div>
<div>Abstract<p><b>Purpose:</b> The success of hematopoietic stem cell transplantation (HSCT) depends on donor cell homing to the bone marrow. However, there is no reliable method of noninvasively monitoring the kinetics and distribution of transferred cells. Using zirconium-89 (<sup>89</sup>Zr)-oxine cell labeling combined with PET imaging, we sought to visualize and quantify donor cell homing in a mouse bone marrow transplantation model.</p><p><b>Experimental Design:</b> The effect of <sup>89</sup>Zr-oxine labeling on bone marrow cell viability and differentiation was evaluated <i>in vitro</i>. <sup>89</sup>Zr-labeled bone marrow cells (2 × 10<sup>7</sup> cells, 16.6 kBq/10<sup>6</sup> cells) were transferred intravenously, and serial microPET images were obtained (<i>n</i> = 5). The effect of a CXCR4 inhibitor, plerixafor (5 mg/kg) and G-CSF (2.5 μg) on bone marrow homing and mobilization were examined (<i>n</i> = 4). Engraftment of the transferred <sup>89</sup>Zr-labeled cells was evaluated (<i>n</i> = 3).</p><p><b>Results:</b> <sup>89</sup>Zr-oxine–labeled bone marrow cells showed delayed proliferation, but differentiated normally. Transferred bone marrow cells rapidly migrated to the bone marrow, spleen, and liver (<i>n</i> = 5). Approximately 36% of donor cells homed to the bone marrow within 4 hours, irrespective of prior bone marrow ablation. Inhibition of CXCR4 by plerixafor alone or with G-CSF significantly blocked the bone marrow homing (<i>P</i> < 0.0001, vs. nontreated, at 2 hours), confirming a crucial role of the CXCR4–CXCL12 system. Mobilization of approximately 0.64% of pretransplanted bone marrow cells induced a 3.8-fold increase of circulating bone marrow cells. <sup>89</sup>Zr-labeled donor cells engrafted as well as nonlabeled cells.</p><p><b>Conclusions:</b> <sup>89</sup>Zr-oxine PET imaging reveals rapid bone marrow homing of transferred bone marrow cells without impairment of their stem cell functions, and thus, could provide useful information for optimizing HSCT. <i>Clin Cancer Res; 23(11); 2759–68. ©2016 AACR</i>.</p></div>
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.