Background: Cytokine-induced killer (CIK) cells are typically differentiated in vitro with interferon (IFN)-gamma and alpha CD3 monoclonal antibodies (mAb), followed by the repeated provision of interleukin (IL)-2. It is presently unknown whether thymoglobulin (TG), a preparation of polyclonal rabbit gamma immunoglobulins directed against human thymocytes, can improve the generation efficiency of CIK cells compared with alpha CD3 mAb in a clinical-grade culture protocol. Methods: Peripheral blood mononuclear cells (PBMC) from 10 healthy donors and 4 patients with solid cancer were primed with IFN-gamma on day 0 and low (50 ng/ml), intermediate (250 ng/ml) and high (500 ng/ml) concentrations of either alpha CD3 mAb or TG on day 1, and were fed with IL-2 every 3 days for 21 days. Aliquots of cells were harvested weekly to monitor the expression of representative members of the killer-like immunoglobulin receptor (KIR), NK inhibitory receptor, NK activating receptor and NK triggering receptor families. We also quantified the frequency of bona fide regulatory T cells (Treg), a T-cell subset implicated in the down-regulation of anti-tumor immunity, and tested the in vitro cytotoxic activity of CIK cells against NK-sensitive, chronic myeloid leukaemia K562 cells. Results: CIK cells expanded more vigorously in cultures supplemented with intermediate and high concentrations of TG compared with 50 ng/ml alpha CD3 mAb. TG-driven CIK cells expressed a constellation of NK activating/inhibitory receptors, such as CD158a and CD158b, NKp46, NKG2D and NKG2A/CD94, released high quantities of IL-12p40 and efficiently lysed K562 target cells. Of interest, the frequency of Treg cells was lower at any time-point compared with PBMC cultures nurtured with alpha CD3 mAb. Cancer patient-derived CIK cells were also expanded after priming with TG, but they expressed lower levels of the NKp46 triggering receptor and NKG2D activating receptor, thus manifesting a reduced ability to lyse K562 cells. Conclusions: TG fosters the generation of functional CIK cells with no concomitant expansion of tumor-suppressive Treg cells. The culture conditions described herein should be applicable to cancer-bearing individuals, although the differentiation of fully functional CIK cells may be hindered in patients with advanced malignancies.

Thymoglobulin, interferon-γ and interleukin-2 efficiently expand cytokine-induced killer (CIK) cells in clinical-grade cultures / Giuseppina, Bonanno; Paola, Iudicone; Andrea, Mariotti; Annabella, Procoli; Annino, Pandolfi; Daniela, Fioravanti; Maria, Corallo; Alessandro, Perillo; Giovanni, Scambia; Pierelli, Luca; Sergio, Rutella. - In: JOURNAL OF TRANSLATIONAL MEDICINE. - ISSN 1479-5876. - 8:1(2010), p. 129. [10.1186/1479-5876-8-129]

Thymoglobulin, interferon-γ and interleukin-2 efficiently expand cytokine-induced killer (CIK) cells in clinical-grade cultures

PIERELLI, LUCA;
2010

Abstract

Background: Cytokine-induced killer (CIK) cells are typically differentiated in vitro with interferon (IFN)-gamma and alpha CD3 monoclonal antibodies (mAb), followed by the repeated provision of interleukin (IL)-2. It is presently unknown whether thymoglobulin (TG), a preparation of polyclonal rabbit gamma immunoglobulins directed against human thymocytes, can improve the generation efficiency of CIK cells compared with alpha CD3 mAb in a clinical-grade culture protocol. Methods: Peripheral blood mononuclear cells (PBMC) from 10 healthy donors and 4 patients with solid cancer were primed with IFN-gamma on day 0 and low (50 ng/ml), intermediate (250 ng/ml) and high (500 ng/ml) concentrations of either alpha CD3 mAb or TG on day 1, and were fed with IL-2 every 3 days for 21 days. Aliquots of cells were harvested weekly to monitor the expression of representative members of the killer-like immunoglobulin receptor (KIR), NK inhibitory receptor, NK activating receptor and NK triggering receptor families. We also quantified the frequency of bona fide regulatory T cells (Treg), a T-cell subset implicated in the down-regulation of anti-tumor immunity, and tested the in vitro cytotoxic activity of CIK cells against NK-sensitive, chronic myeloid leukaemia K562 cells. Results: CIK cells expanded more vigorously in cultures supplemented with intermediate and high concentrations of TG compared with 50 ng/ml alpha CD3 mAb. TG-driven CIK cells expressed a constellation of NK activating/inhibitory receptors, such as CD158a and CD158b, NKp46, NKG2D and NKG2A/CD94, released high quantities of IL-12p40 and efficiently lysed K562 target cells. Of interest, the frequency of Treg cells was lower at any time-point compared with PBMC cultures nurtured with alpha CD3 mAb. Cancer patient-derived CIK cells were also expanded after priming with TG, but they expressed lower levels of the NKp46 triggering receptor and NKG2D activating receptor, thus manifesting a reduced ability to lyse K562 cells. Conclusions: TG fosters the generation of functional CIK cells with no concomitant expansion of tumor-suppressive Treg cells. The culture conditions described herein should be applicable to cancer-bearing individuals, although the differentiation of fully functional CIK cells may be hindered in patients with advanced malignancies.
2010
methods; cells; killer cells; neoplasms; cytotoxicity; cytology/drug effects; pharmacology; antilymphocyte serum; young adult; immunology; regulatory; cytokine-induced killer cells; cell culture techniques; interleukin-2; animals; rabbits; middle aged; humans; phenotype; adult; drug effects; cell proliferation; female; t-lymphocytes; interferon-gamma; tissue donors; cultured; cd8-positive t-lymphocytes; natural; immunologic
01 Pubblicazione su rivista::01a Articolo in rivista
Thymoglobulin, interferon-γ and interleukin-2 efficiently expand cytokine-induced killer (CIK) cells in clinical-grade cultures / Giuseppina, Bonanno; Paola, Iudicone; Andrea, Mariotti; Annabella, Procoli; Annino, Pandolfi; Daniela, Fioravanti; Maria, Corallo; Alessandro, Perillo; Giovanni, Scambia; Pierelli, Luca; Sergio, Rutella. - In: JOURNAL OF TRANSLATIONAL MEDICINE. - ISSN 1479-5876. - 8:1(2010), p. 129. [10.1186/1479-5876-8-129]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/385812
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