title: Surface biotinylation of cytotoxic T lymphocytes for in vivo tracking of tumor immunotherapy in murine models.
abstract: 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 with89Zr-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.
authors.affiliation: Department of Radiology, Huashan Hospital, Fudan University, 12 Urumchi Road, Shanghai, 200040, China.|Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.|Department of Radiology, Qilu Hospital of Shandong University, 107 West Wenhua Road, Jinan, 250012, China.
authors.forename: Anning
authors.initials: A
authors.lastname: Li
authors.affiliation: Department of Radiology, Huashan Hospital, Fudan University, 12 Urumchi Road, Shanghai, 200040, China.|Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Yue
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Jenny
authors.initials: J
authors.lastname: Linnoila
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.|Department of Radiology, Institute for Innovation in Imaging, Massachusetts General Hospital, Boston, MA, 02114, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Pulli
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Cuihua
authors.initials: C
authors.lastname: Wang
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Matthias
authors.initials: M
authors.lastname: Zeller
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Ali
authors.affiliation: Molecular Neurogenetics Unit, Neuroscience Center, 149 13th St., Charlestown, MA, 02129, USA.
authors.forename: Grant K
authors.initials: GK
authors.lastname: Lewandrowski
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Jinghui
authors.initials: J
authors.lastname: Li
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Benoit
authors.initials: B
authors.lastname: Tricot
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Edmund
authors.initials: E
authors.lastname: Keliher
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA.
authors.forename: Gregory R
authors.initials: GR
authors.lastname: Wojtkiewicz
authors.affiliation: Brain Tumor Research Center, Simches Research Building, Neurosurgery Service, Massachusetts General Hospital, Boston, MA, 02114, USA.
authors.forename: Giulia
authors.initials: G
authors.lastname: Fulci
authors.affiliation: Department of Radiology, Huashan Hospital, Fudan University, 12 Urumchi Road, Shanghai, 200040, China.
authors.forename: Xiaoyuan
authors.initials: X
authors.lastname: Feng
authors.affiliation: Molecular Neurogenetics Unit, Neuroscience Center, 149 13th St., Charlestown, MA, 02129, USA.
authors.forename: Bakhos A
authors.initials: BA
authors.lastname: Tannous
authors.affiliation: Department of Radiology, Huashan Hospital, Fudan University, 12 Urumchi Road, Shanghai, 200040, China. aocnhnr@126.com.
authors.forename: Zhenwei
authors.initials: Z
authors.lastname: Yao
authors.affiliation: Center for Systems Biology, Massachusetts General Hospital and Harvard Medical School, 185 Cambridge Street, Boston, MA, 02114, USA. jwchen@mgh.harvard.edu.|Department of Radiology, Institute for Innovation in Imaging, Massachusetts General Hospital, Boston, MA, 02114, USA. jwchen@mgh.harvard.edu.
authors.forename: John W
authors.initials: JW
authors.lastname: Chen
mesh_terms: D000818:Animals; D019921:Biotinylation / Q000379:methods*; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D051379:Mice; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism*
keywords: Biotin; Cytotoxic T lymphocytes; Immunotherapy; Tumor