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Tumor-infiltrating immune repertoires captured by single-cell barcoding in emulsion

By Adrian W Briggs, Stephen J Goldfless, Sonia Timberlake, Brian J Belmont, Christopher R Clouser, David Koppstein, Devin Sok, Jason Vander A Heiden, Manu V Tamminen, Steven H Kleinstein, Dennis R Burton, George M Church, Francois Vigneault

Posted 05 May 2017
bioRxiv DOI: 10.1101/134841

Tumor-infiltrating lymphocytes (TILs) are critical to anti-cancer immune responses, but their diverse phenotypes and functions remain poorly understood and challenging to study. We therefore developed a single-cell barcoding technology for deep characterization of TILs without the need for cell-sorting or culture. Our emulsion-based method captures full-length, natively paired B-cell and T-cell receptor (BCR and TCR) sequences from lymphocytes among millions of input cells. We validated the method with 3 million B-cells from healthy human blood and 350,000 B-cells from an HIV elite controller, before processing 400,000 cells from an unsorted dissociated ovarian adenocarcinoma and recovering paired BCRs and TCRs from over 11,000 TILs. We then extended the barcoding method to detect DNA-labeled antibodies, allowing ultra-high throughput, simultaneous protein detection and RNA sequencing from single cells.

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