A Near-Infrared Photoimmunotherapy Technique to Treat Lung Cancer in a Murine Model
Published: April 30, 2024
Abstract
Source: Yasui, H., et al. Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination. J. Vis. Exp. (2021)
This video demonstrates a near-infrared photoimmunotherapy technique to treat lung cancer in an engineered mouse model exhibiting luciferase-expressing tumor cells. Upon injecting an antibody-photosensitizer conjugate (APC) that binds to specific antigens on the tumor cells, the mouse is exposed to near-infrared radiation, inducing photochemical changes in the APC and leading to necrotic death of the bound tumor cells. The effectiveness of the treatment is monitored by injecting luciferin, a luciferase substrate, and measuring the bioluminescence over time, correlating with tumor growth.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Conjugation of IR700 with mAb Incubate monoclonal antibody (mAb) (1 mg, 6.8 nmol) with IR700 N-Hydroxysuccinimide (NHS) ester (66.8 mg, 34.2 nmol, 5 mmol/L in dimethyl sulfoxide [DMSO]) in 0.1 mol/L Na2HPO4 (pH 8.6) at 15-25 °C for 1 h. Purify the mixture using a column (e.g., Sephad…
Representative Results
Figure 1: Easy hand-made device for cell transplantation. Attach the stopper made with polystyrene foam to the 30G needle so that the tip remains at 5 mm. The tip of the needle should be bent to avoid pneumothorax. Figure 2: Injection of target cells in…
Divulgaciones
The authors have nothing to disclose.
Materials
| 0.25w/v% Trypsin-1mmol/l EDTA 4Na Solution with Phenol Red | Wako | 209-016941 | for cell culture |
| 1mL syringe | TERUMO | SS-01T | for mice experiment |
| 30G needle | Nipro | 1907613 | for mice experiment |
| BALB/cSlc-nu/nu | Japan SLC | ||
| Collidal Blue Staining Kit | Invitrogen | LC6025 | use for gel protein staining |
| Coomassie (bradford) Plus protein assay | Thermo Fisher Scientific Inc (Waltham, MA, USA) | PI-23200 | for measuring the APC concentration |
| Dimethyl sulfoxide (DMSO) | Wako | 043-07216 | use for conjugation of IR700 |
| D-Luciferin (potassium salt) | Cayman Chemical | 14681 | for bioluminescence imaging and DLIT |
| GraphPad Prism7 | GraphPad software | for statistical analysis | |
| Image Studio | Li-Cor Biosciences | for analyzing 700 nm fluorescent image | |
| IRDye 700DX Ester Infrared Dye | LI-COR Bioscience (Lincoln, NE, USA) | 929-70011 | |
| Isoflurane | Wako | 095-06573 | for mice anesthesia |
| IVIS Spectrum CT | PerkinElmer | for capturing bioluminescent image and DLIT | |
| Living Image | PerkinElmer | for analyzing bioluminescent image and DLIT | |
| Na2HPO4 | SIGMA-ALDRICH (St. Louis, MO, USA) | S9763 | use for conjugation of IR700 |
| NIR Laser | Changchun New Industries Optoelectronics Technology | MRL-III-690R | for NIR irradiation |
| Novex WedgeWell 4 to 20%, Tris-Glycine, 1.0 mm, Mini Protein Gel, 12 well | Invitrogen | XP04202BOX | use for SDS-PAGE |
| NuPAGE LDS Sample Buffer (x4) | Invitrogen | NP0007 | use for SDS-PAGE |
| Optical power meter | Thorlabs (Newton, NJ, USA) | PM100 | for measuring the output of the NIR laser |
| PBS(-) | Wako | 166-23555 | |
| Pearl Trilogy imaging system | Li-Cor Biosciences | for capturing 700 nm fluorecent image | |
| Penicilin-Streptomycin Solution (x100) | Wako | 168-23191 | for cell culture |
| Puromycin Dihydrochloride | ThermoFisher | A1113803 | for luciferase transfection |
| RediFect Red-Fluc-Puromycin Lentiviral Prticles | PerkinElmer | CLS960002 | for luciferase transfection |
| RPMI-1640 with L-glutamine and Phenol Red | Wako | 189-02025 | for cell culture |
| Sephadex G25 column (PD-10) | GE Healthcare (Piscataway, NJ, USA) | 17-0851-01 | use for conjugation of IR700 |
| UV-1900i | Shimadzu | for measuring the APC concentration |
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Citar este artículo
A Near-Infrared Photoimmunotherapy Technique to Treat Lung Cancer in a Murine Model. J. Vis. Exp. (Pending Publication), e22225, doi: (2024).