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Molecular Medicine

, Volume 23, Issue 1, pp 235–246 | Cite as

Analytic and Dynamic Secretory Profile of Patient-Derived Cytokine-lnduced Killer Cells

  • Giulia Mesiano
  • Roberta Zini
  • Giulia Montagner
  • Nicoletta Bianchi
  • Rossella Manfredini
  • Antonella Chillemi
  • Massimo Aglietta
  • Giovanni Grignani
  • Ilaria Lampronti
  • Erika Fiorino
  • Fabio Malavasi
  • Dario Sangiolo
  • Roberto Gambari
  • Davide Ferrari
Research Article

Abstract

Adoptive immunotherapy with cytokine induced killer (CIK) cells has shown antitumor activity against several kinds of cancer in preclinical models and clinical trials. CIK cells are a subset of ex vivo expanded T lymphocytes with T-NK phenotype and MHC-unrestricted antitumor activity. The literature provides scant information on cytokines, chemokines and growth factors secreted by CIK cells. Therefore, we investigated the secretory profile of CIK cells generated from tumor patients. The secretome analysis was performed at specific time points (d 1, d 14 and d 21) of CIK cell expansion. Mature CIK cells (d 21) produce a great variety of interleukins and secreted proteins that can be divided into three groups based on their secretion quantity: high (interleukin [IL]-13, regulated on activation normal T cell expressed and secreted [RANTES] chemokine, MIP-1α and 1β), medium (IL-1Ra, IL-5, IL-8, IL-10, IL-17, IP-10, INF-γ, vascular endothelial growth factor [VEGF] and granulocyte-macrophage colony-stimulating factor [GM-CSF]) and low (IL-1β, IL-4, IL-6, IL-7, IL-9, IL-12, IL-15, eotaxin, platelet-derived growth factor-bb, basic fibroblast growth factor, G-CSF and monocyte chemoattractant protein [MCP]-1). Moreover, comparing peripheral blood mononuclear cells (PBMCs) (d 1) and mature CIK cells (d 14 and 21) secretomes, we observed that IL-5, IL-10, IL-13, GM-CSF and VEGF were greatly upregulated, while IL-1β, IL-6, IL-8, IL-15, IL-17, eotaxin, MCP-1 and RANTES were downregulated. We also performed a gene expression profile analysis of patient-derived CIK cells, showing that mRNA for the different cytokines and secreted proteins was modulated during PBMC-to-CIK differentiation. We highlight previously unknown secretory properties and provide, for the first time, a comprehensive molecular characterization of CIK cells. Our findings provide a rationale to explore the functional implications and possible therapeutic modulation of CIK secretome.

Notes

Acknowledgments

This work was supported in part by FPRC ONLUS 5 × 1000, Ministero della Salute 2012; Associazione Italiana per la Ricerca sul Cancro (AIRC), project no. 10005 Special Program Molecular Clinical Oncology 5 × 1000 to AIRC-Gruppo Italiano Malattie Mieloproliferative, AIRC project no. 15337. DS is the recipient of a grant from Ricerca Finalizzata-Giovani Ricercatori Ministero della Salute (GR-2011-02349197).

Supplementary material

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Authors and Affiliations

  • Giulia Mesiano
    • 1
  • Roberta Zini
    • 3
  • Giulia Montagner
    • 4
  • Nicoletta Bianchi
    • 4
  • Rossella Manfredini
    • 3
  • Antonella Chillemi
    • 5
  • Massimo Aglietta
    • 1
    • 2
  • Giovanni Grignani
    • 1
    • 2
  • Ilaria Lampronti
    • 4
  • Erika Fiorino
    • 2
  • Fabio Malavasi
    • 5
  • Dario Sangiolo
    • 1
    • 2
  • Roberto Gambari
    • 4
  • Davide Ferrari
    • 4
  1. 1.Division of Medical Oncology, Experimental Cell TherapyCandiolo Cancer Institute, FPO-IRCCSTorinoItaly
  2. 2.Department of OncologyUniversity of TorinoTorinoItaly
  3. 3.Centre for Regenerative Medicine “Stefano Ferrari,” Department of Life SciencesUniversity of Modena and Reggio EmiliaModenaItaly
  4. 4.Department of Life Science and Biotechnology, Sections of Microbiology and Applied Pathology; Biochemistry and Molecular BiologyUniversity of FerraraFerraraItaly
  5. 5.Laboratory of Immunogenetics and CeRMS, Department of Medical SciencesUniversity of TorinoTorinoItaly

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