Differentiation of glioma and radiation injury in rats using in vitro produce magnetically labeled cytotoxic T-cells and MRI

• Published in: PloS one 2010-02, Vol.5 (2), p.e9365
• Main Authors: Arbab, Ali S, Janic, Branislava, Jafari-Khouzani, Kourosh, Iskander, A S M, Kumar, Sanath, Varma, Nadimpalli R S, Knight, Robert A, Soltanian-Zadeh, Hamid, Brown, Stephen L, Frank, Joseph A
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Blood; Brain cancer; Brain Neoplasms - immunology; Brain Neoplasms - pathology; Brain Neoplasms - therapy; Brain tumors; Cancer therapies; CAT scans; CD14 antigen; CD25 antigen; CD4 antigen; CD4 Antigens - immunology; CD8 antigen; CD8 Antigens - immunology; CD83 antigen; CD86 antigen; Cell death; Cell Line, Tumor; Computed tomography; Cord blood; Cytokines; Cytotoxicity; Data processing; Dendritic cells; Dendritic Cells - cytology; Dendritic Cells - immunology; Diagnosis, Differential; Fetal Blood - cytology; Glioma; Glioma - immunology; Glioma - pathology; Glioma - therapy; Glioma cells; Gliomas; Histocompatibility antigen HLA; Hospitals; Humans; Identification methods; Image acquisition; Immunology/Antigen Processing and Recognition; Immunotherapy, Adoptive - methods; In vitro methods and tests; Injuries; Interleukin-2 Receptor alpha Subunit - immunology; Iron; L-Lactate dehydrogenase; Labeling; Laboratories; Lactate dehydrogenase; Ligands; Lymphocytes T; Magnetic resonance; Magnetic resonance imaging; Magnetic Resonance Imaging - methods; Magnetics; Medical imaging; Medical prognosis; Nanoparticles; Necrosis; Neoplasms - etiology; Neoplasms - immunology; Neoplasms - therapy; Neoplasms, Radiation-Induced - immunology; Neoplasms, Radiation-Induced - pathology; Neoplasms, Radiation-Induced - therapy; NMR; Nuclear magnetic resonance; Oncology; Oncology/Neuro-Oncology; Permeability; Positron emission; Probes; Radiation; Radiation (Physics); Radiation injuries; Radiation therapy; Radiology and Medical Imaging/Magnetic Resonance Imaging; Radiotherapy; Radiotherapy - adverse effects; Rats; Rodents; Spectrum analysis; Spinal cord injuries; Stem cells; Studies; Surgery; T cells; T-Lymphocytes, Cytotoxic - cytology; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - transplantation; Tomography; Toxicity; Tumors; Vaccination; Xenograft Model Antitumor Assays - methods
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0009365

A limitation with current imaging strategies of recurrent glioma undergoing radiotherapy is that tumor and radiation injury cannot be differentiated with post contrast CT or MRI, or with PET or other more complex parametric analyses of MRI data. We propose to address the imaging limitation building on emerging evidence indicating that effective therapy for recurrent glioma can be attained by sensitized T-cells following vaccination of primed dendritic cells (DCs). The purpose of this study was to determine whether cord blood T-cells can be sensitized against glioma cells (U-251) and if these sensitized cytotoxic T-cells (CTLs) can be used as cellular magnetic resonance imaging probes to identify and differentiate glioma from radiation necrosis in rodent models. Cord blood T and CD14+ cells were collected. Isolated CD14+ cells were then converted to dendritic cells (DCs), primed with glioma cell lysate and used to sensitize T-cells. Phenotypical expression of the generated DCs were analyzed to determine the expression level of CD14, CD86, CD83 and HLA-DR. Cells positive for CD25, CD4, CD8 were determined in generated CTLs. Specificity of cytotoxicity of the generated CTLs was also determined by lactate dehydrogenase (LDH) release assay. Secondary proliferation capacity of magnetically labeled and unlabeled CTLs was also determined. Generated CTLs were magnetically labeled and intravenously injected into glioma bearing animals that underwent MRI on days 3 and 7 post- injection. CTLs were also administered to animals with focal radiation injury to determine whether these CTLs accumulated non-specifically to the injury sites. Multi-echo T2- and T2*-weighted images were acquired and R2 and R2* maps created. Our method produced functional, sensitized CTLs that specifically induced U251 cell death in vitro. Both labeled and unlabeled CTLs proliferated equally after the secondary stimulation. There were significantly higher CD25 positive cells (p =