Volume 13 Supplement 1

Melanoma Bridge 2014: meeting abstracts

Open Access

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

  • Rossana Tallerico1,
  • Costanza M Cristiani1,
  • Mariaelena Capone2,
  • Gabriele Madonna2,
  • Domenico Mallardo2,
  • Ester Simeone2,
  • Andrea Dominijanni3,
  • Antonio M Grimaldi2,
  • Francesco Colucci4,
  • Paolo A Ascierto2 and
  • Ennio Carbone1, 5
Journal of Translational Medicine201513(Suppl 1):O8

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

Published: 15 January 2015

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.

Authors’ Affiliations

(1)
Tumorimmunology and Immunopathology Laboratory, Department of Experimental and Clinic Medicine, University "Magna Graecia" of Catanzaro
(2)
Melanoma. CancerImmunotherapy and Innovative Therapy Unit, Istituto Nazionale Tumori Fondazione "G. Pascale"
(3)
Immunohaematological Transfusion Medicine Centre “Pugliese-Ciaccio” Hospital of Catanzaro
(4)
Department of Obstetrics and Gynaecology, University of Cambridge Clinical School
(5)
Department of Microbiology Cell and Tumorbiology (MTC), Karolinska Institutet

References

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Copyright

© Tallerico et al; licensee BioMed Central Ltd. 2015

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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