Skip to main content
  • Oral presentation
  • Open access
  • Published:

Analysis of T and NK cells immune response in Ipilimumab treated Melanoma patients

Background

The most promising immunotherapeutics tested in metastatic melanoma patients are the monoclonal antibody blocking CTLA-4 (Ipilimumab), and those interfering with PD-1 and PD-L1. However, the lack of knowledge on predictive biomarkers that could assist the treatment remains a limiting factor. We speculate that, along with additional markers, the immunoscore [1] is fundamental as prognostic and predictive marker for response to immunotherapies in metastatic melanoma. Our previous data demonstrate that NK cells control the melanoma progression [2, 3]. Therefore we have analysed both T cells and NK cells subsets frequencies and receptors repertoire in the peripheral blood of Ipilimumab treated patients with Stage IV metastatic melanoma.

Material and methods

Peripheral blood mononuclear cells (PBMCs) from 12 different patients with stage IV metastatic melanoma were collected and analyzed. Each patient received 4 infusions of Ipilimumab each 21 days. Before each infusion we collected patients’ blood and isolated PBMCs to analyze the lymphocytes compartment.

We stained for the following antibodies: CD56 PE, CD3 Fitc, CD56 APC, CD4 PeCy7, CD8 APCCy7, CD152 (CTLA4) PE, CD279 (PD-1) PE, CCR7 PeCy7, CD158a/h(KIR2DL1/S1) PE, CD158b (KIR2DL2/DL3) PE, CD158e (KIR3DL1) PE, CD16 APCCy7, CD57 PE, CD69 PE, CD314 (NKG2D) PE, CD226 (DNAM-1) PE, CD337 (NKp30) PE, CD336 (NKp44) PE, CD335 (NKp46) PE (Miltenyi Biotech), CD192 (CCR2) AlexaFluor 647, CXCR2 (IL8RB) APC, 7-AADStaining Solution (BD Italia), TIM3 PE (ebioscience), NKG2C PE (R&D Systems). The analysis was performed with FACS CANTO II. Statistical analysis was performed with Anova and Student’s t-test.

Results

Our data indicate that, after the first Ipilimumab treatment, an inversion of CD4/CD8 ratio occurs with a concomitant increase in the CD56dim population and a higher expression of TIM-3 and NKp46 molecules on the surface of NK cells. Moreover, the frequency of NK and T cells expressing KIRs and CCR7 is reduced, while the mean fluorescence intensity of CD16 and PD1 is upregulated on both CD56bright and CD56dim NK cells.

Conclusions

These preliminary data indicate that early during Ipilimumab treatment, cytotoxic lymphocytes CD8+ T cells and CD56dim NK cells expand and become activated. NK cells seem to be polarized towards a CD56dimCD16bright KIRs+NKp46+TIM3+ phenotype. Ipilimumab treatment may induce NK cells maturation, which might in turn drive activation of CD8+ T cells.

References

  1. Galon J, Pagès F, Marincola FM, Thurin M, Trinchieri G, Fox BA, Gajewski TF, Ascierto PA: The immune score as a new possible approach for the classification of cancer. J Transl Med. 2012, 10: 1-10.1186/1479-5876-10-1.

    Article  PubMed Central  PubMed  Google Scholar 

  2. Lakshmikanth T, Burke S, Ali TH, Kimpfler S, Ursini F, Ruggeri L, Capanni M, Umansky V, Paschen A, Sucker A, Pende D, Groh V, Biassoni R, Höglund P, Kato M, Shibuya K, Schadendorf D, Anichini A, Ferrone S, Velardi A, Kärre K, Shibuya A, Carbone E, Colucci F: NCRs and DNAM-1 mediate NK cell recognition and lysis of human and mouse melanoma cell lines in vitro and in vivo. J Clin Invest. 2009, 119 (5): 1251-1263. 10.1172/JCI36022.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Burke S, Lakshmikanth T, Colucci F, Carbone E: New views on natural killer cell-based immunotherapy for melanoma treatment. Trends Immunol. 2010, 31 (9): 339-345. 10.1016/j.it.2010.06.003.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Open Access  This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.

The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this licence, visit https://creativecommons.org/licenses/by/4.0/.

The Creative Commons Public Domain Dedication waiver (https://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tallerico, R., Cristiani, C.M., Capone, M. et al. Analysis of T and NK cells immune response in Ipilimumab treated Melanoma patients. J Transl Med 13 (Suppl 1), O8 (2015). https://doi.org/10.1186/1479-5876-13-S1-O8

Download citation

  • Published:

  • DOI: https://doi.org/10.1186/1479-5876-13-S1-O8

Keywords