Surface biotinylation of cytotoxic T lymphocytes for in vivo tracking of tumor immunotherapy in murine models

生物素 体内 生物素化 细胞毒性T细胞 CD8型 免疫疗法 癌症研究 化学 胶质瘤 CTL公司* 链霉亲和素 分子生物学 体外 免疫学 医学 生物 免疫系统 生物化学 生物技术
作者
Anning Li,Yue Wu,Jenny Linnoila,Benjamin Pulli,Cuihua Wang,Matthias W. G. Zeller,Muhammad Ali,Grant K. Lewandrowski,Jinghui Li,Benoit Tricot,Edmund J. Keliher,Gregory R. Wojtkiewicz,Giulia Fulci,Xiaoyuan Feng,Bakhos A. Tannous,Zhenwei Yao,John W. Chen
出处
期刊:Cancer Immunology, Immunotherapy [Springer Science+Business Media]
卷期号:65 (12): 1545-1554 被引量:10
标识
DOI:10.1007/s00262-016-1911-9
摘要

Currently, there is no stable and flexible method to label and track cytotoxic T lymphocytes (CTLs) in vivo in CTL immunotherapy. We aimed to evaluate whether the sulfo-hydroxysuccinimide (NHS)-biotin–streptavidin (SA) platform could chemically modify the cell surface of CTLs for in vivo tracking. CD8+ T lymphocytes were labeled with sulfo-NHS-biotin under different conditions and then incubated with SA–Alexa647. Labeling efficiency was proportional to sulfo-NHS-biotin concentration. CD8+ T lymphocytes could be labeled with higher efficiency with sulfo-NHS-biotin in DPBS than in RPMI (P < 0.05). Incubation temperature was not a key factor. CTLs maintained sufficient labeling for at least 72 h (P < 0.05), without altering cell viability. After co-culturing labeled CTLs with mouse glioma stem cells (GSCs) engineered to present biotin on their surface, targeting CTLs could specifically target biotin-presenting GSCs and inhibited cell proliferation (P < 0.01) and tumor spheres formation. In a biotin-presenting GSC brain tumor model, targeting CTLs could be detected in biotin-presenting gliomas in mouse brains but not in the non-tumor-bearing contralateral hemispheres (P < 0.05). In vivo fluorescent molecular tomography imaging in a subcutaneous U87 mouse model confirmed that targeting CTLs homed in on the biotin-presenting U87 tumors but not the control U87 tumors. PET imaging with 89Zr-deferoxamine-biotin and SA showed a rapid clearance of the PET signal over 24 h in the control tumor, while only minimally decreased in the targeted tumor. Thus, sulfo-NHS-biotin–SA labeling is an efficient method to noninvasively track the migration of adoptive transferred CTLs and does not alter CTL viability or interfere with CTL-mediated cytotoxic activity.
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