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…

Divulgations

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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Citer Cet Article
A Near-Infrared Photoimmunotherapy Technique to Treat Lung Cancer in a Murine Model. J. Vis. Exp. (Pending Publication), e22225, doi: (2024).

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