Source: Liu, H. Y., et al. Experimental Melanoma Immunotherapy Model Using Tumor Vaccination with a Hematopoietic Cytokine. J. Vis. Exp. (2023).
This video demonstrates a cell-based tumor vaccination assay. A mouse, intradermally injected with B16-F10 melanoma cells for tumor formation, is vaccinated with B16-F10 cells secreting Flt3L cytokine near the developed tumor. The secreted Flt3L from the vaccine promotes the maturation and proliferation of dendritic cells that then present tumor antigens to T cells in lymph nodes, triggering a targeted anti-tumor immune response.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
All mice used in the study were maintained and housed in the vivarium of the La Jolla Institute for Immunology (LJI) under specific pathogen-free conditions with controlled temperature and humidity. Animal experiments were performed with 8-14 weeks-old female C57BL/6 mice according to guidelines and protocols approved by the LJI Animal Care Committee.
1. Preparation of cultured tumor cells for implantation
2. Tumor implantation
3. Vaccine preparation and injection of Flt3L-expressing B16-F10 (B16-Flt3L) cells
The authors have nothing to disclose.
0.25% trypsin-EDTA | Gibco | 25200-056 | |
10% heat-inactivated FBS | Omega Scientific | FB-02 | Lot# 209018 |
30G needle | BD Biosciences | 305106 | |
B16 cell line expressing Fms-like tyrosine kinase 3 ligand (B16-Flt3L) | Gift of Dr. Stephen Schoenberger, LJI | Flt3L cDNAs were cloned into the pMG-Lyt2 retroviral vector | |
B16-F10 cell lines | ATCC | CRL-6475 | |
Centrifuge 5810R | Eppendorf | ||
Dulbecco's Modified Eagle Medium (DMEM) | Corning | 10013CV | |
Electronic digital caliper | Fisherbrand | 14-648-17 | |
Iscove's modified Dulbecco's medium (IMDM) | Thermo Fisher | 12440053 | |
MEM nonessential amino acids | Gibco | 11140-050 | |
penicillin and streptomycin | Gibco | 15140-122 | |
RPMI 1640 medium | Corning | 10-040-CV | |
RS2000 X-ray Irradiator | Rad Source Technologies | ||
sodium pyruvate | Gibco | 11360-070 |
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