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  • A novel system to map protein interactions reveals evolutionarily conserved immune evasion pathways on transmissible cancers.

A novel system to map protein interactions reveals evolutionarily conserved immune evasion pathways on transmissible cancers.

Science advances (2020-09-17)
Andrew S Flies, Jocelyn M Darby, Patrick R Lennard, Peter R Murphy, Chrissie E B Ong, Terry L Pinfold, Alana De Luca, A Bruce Lyons, Gregory M Woods, Amanda L Patchett
ABSTRACT

Around 40% of humans and Tasmanian devils (Sarcophilus harrisii) develop cancer in their lifetime, compared to less than 10% for most species. In addition, devils are affected by two of the three known transmissible cancers in mammals. Immune checkpoint immunotherapy has transformed human medicine, but a lack of species-specific reagents has limited checkpoint immunology in most species. We developed a cut-and-paste reagent development system and used the fluorescent fusion protein system to show that immune checkpoint interactions are conserved across 160,000,000 years of evolution, CD200 is highly expressed on transmissible tumor cells, and coexpression of CD200R1 can block CD200 surface expression. The system's versatility across species was demonstrated by fusing a fluorescent reporter to a camelid-derived nanobody that binds human programmed death ligand 1. The evolutionarily conserved pathways suggest that naturally occurring cancers in devils and other species can be used to advance our understanding of cancer and immunological tolerance.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
TEV Protease
Sigma-Aldrich
RPMI-1640 Medium, Modified, with sodium bicarbonate, without L-glutamine and phenol red, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Hygromycin B solution from Streptomyces hygroscopicus, ≥60% (HPLC), 45-60 mg/mL in H2O
SAFC
EX-CELL® CD CHO Serum-Free Medium for CHO Cells, Chemically Defined, Animal-component free, with hypoxanthine, with thymidine, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Histopaque®-1077, sterile-filtered, density: 1.077 g/mL
Sigma-Aldrich
Pur-A-Lyzer Maxi Dialysis Kit, Maxi 6000, capacity 0.1-3 mL, pkg of 5 Maxi dialysis tubes, MWCO 6-8 kDa
TPP® TubeSpin bioreactor tubes, capacity 50 mL
Sigma-Aldrich
DAPI, for nucleic acid staining