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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)

Background

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.

Methods

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.

Results

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.

Table 1 Therapy-associated T cell clonal expansion in TILs and PBMCs.

Conclusion

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.

References

  1. 1.

    Waitz R, Solomon S: Potent induction of tumor immunity by combining tumor cryoablation with anti-CTLA-4 therapy. Cancer Res. 72: 430-9.

  2. 2.

    Diab A, McArthur H: A pilot study of preoperative (Pre-op), single-dose Ipilimumab (Ipi) and/or cryoablation (Cryo) in women (pts) with early-stage/resectable breast cancer (ESBC). ASCO Annual Meeting. 2014, Abs #1098, Chicago, IL

  3. 3.

    Page D, Yuan J: T cell receptor (TCR) DNA deep sequencing to evaluate clonality of tumor-infiltrating lymphocytes (TILs) in early-stage breast cancer patients (pts) receiving preoperative cryoablation (cryo) and/or Ipilimumab (Ipi). ASCO Annual Meeting. 2014, Chicago, IL, , Abs #3021

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Author information

Correspondence to David B Page.

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This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Keywords

  • Peripheral Blood Mononuclear Cell
  • Cell Clone
  • Ipilimumab
  • Deep Sequencing
  • Early Stage Breast Cancer