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Matched T cell repertoire analysis of peripheral blood and tumor-infiltrating lymphocytes (TILs) in early stage breast cancer (ESBC) patients (pts) treated with pre-operative cryoablation (cryo) and/or Ipilimumab (Ipi)
  1. David B Page1,
  2. Heather McArthur1,
  3. Zhiwan Dong2,
  4. Phillip Wong1,
  5. Ryan Emerson3,
  6. Zhenyu Mu2,
  7. Chunjun Zhao2,
  8. Christopher Comstock1,
  9. Elizabeth Morris1,
  10. Elizabeth Comen1,
  11. Alan Kotin1,
  12. Janice Sung1,
  13. Edi Brogi1,
  14. Monica Morrow1,
  15. Stephen Solomon1,
  16. Virgilio Sacchini1,
  17. Majid Maybody1,
  18. Deirdre Neville1,
  19. Adi Diab4,
  20. Padmanee Sharma4,
  21. Harlan Robins5,
  22. Sujata Patil1,
  23. Jedd D Wolchok1,
  24. Clifford Hudis1,
  25. Larry Norton1,
  26. James Allison4 and
  27. Jianda Yuan6
  1. Aff1 grid.51462.340000000121719952Memorial Sloan Kettering Cancer Center New York NY USA
  2. Aff2 grid.51462.340000000121719952Sloan Kettering Institute New York NY USA
  3. Aff3 grid.421940.aAdative Biotechnologies Seattle WA USA
  4. Aff4 grid.240145.60000000122914776MD Anderson Cancer Center Houston TX USA
  5. Aff5 grid.270240.30000000121801622Fred Hutchinson Cancer Center Seattle WA USA
  6. Aff6 grid.51462.340000000121719952Immune Monitoring FacilitySloan Kettering Institute New York NY USA

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Meeting abstracts


Cryo plus anti-CTLA-4 therapy induces antigen-specific clonal T cell expansion, enhanced survival, and long-term anti-tumor immunity in mice [1]. We recently demonstrated that pre-operative cryo and/or anti-CTLA-4 therapy with Ipi is well tolerated and clinically feasible in women with ESBC. Furthermore, cryo with or without Ipi generates a polyclonal influx of novel T cell clones within the tumor bed [2, 3]. Here, we utilize T cell repertoire analysis to explore the impact of cryo and/or Ipi on clonal expansion within peripheral blood and TILs.


In a pilot study, women with ESBC were treated with cryo 7-10d before mastectomy (6 pts), single-dose Ipi (10 mg/kg) 8-15d before mastectomy (6 pts), or cryo+Ipi (6 pts). Peripheral blood mononuclear cells (PBMCs) and tumor tissue were obtained pre-mastectomy (immediately preceding cryo and/or 1-5d after Ipi), and at mastectomy. T cell repertoire analysis was conducted on extracted DNA using an Illumina® DNA deep sequencing platform and ImmunoSEQ™ software. Clones comprising ≥0.01% of sample DNA were analyzed, and results are reported descriptively.


Cryo with or without Ipi was associated with decreases in absolute TIL count (median change: Ipi +6%, cryo -73%, cryo+Ipi -16%). However, cryo+Ipi was associated with the greatest expansion of TIL clones across the range of 102-104 amplicons (table 1), although no difference was observed by group in PBMC clones. Across all samples, a median of 523 TIL clones increased by ≥102 amplicons, and a median of 4 TIL clones increased by ≥103 amplicons. The Ipi/cryo group exceeded the median in 80% (4/5) of cases. 21% of all TIL clones were detectable in time-matched PBMC, whereas 16% of expanding (≥102) TIL clones were detectable in time-matched PBMC.

Therapy-associated T cell clonal expansion in TILs and PBMCs.


Cryo plus Ipi expands more TIL clones than either strategy alone. Therapy-associated clonal expansion may be difficult to detect in PBMCs. These data highlight the potential importance of TIL repertoire analysis for the monitoring of pts treated with cryo and/or Ipi in the preoperative setting. In a follow-up randomized study, we will evaluate whether TIL clonal expansion across the 102-104 range can be used to predict recurrence-free survival.


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