Dendritic cell-activated cytokine-induced killer cells enhance the anti-tumor effect of chemotherapy on non-small cell lung cancer in patients after surgery

doi:10.3109/14653240903121252

Cytotherapy

Volume 11, Issue 8, 2009, Pages 1076-1083

https://doi.org/10.3109/14653240903121252 Get rights and content

Background aims

Cytokine-induced killer (CIK) cells have shown cytolytic activity against several tumor cells in vitro and in animal tumor models. Furthermore, CIK cells activated by dendritic cell (DC) stimulation show increased anti-tumor activity. This study aimed to evaluate the clinical efficacy of DC-activated CIK cell treatment following regular chemotherapy and the effects of this therapy on immune responses in patients with non-small cell lung cancer (NSCLC) after surgery.

Methods

A paired study, with 42 patients in each group with stage I–IIIa NSCLC after surgery, was performed. Patients received chemotherapy alone (CT) or chemotherapy and DC-activated CIK cell treatment (immuno-CT). Disease-free survival (DFS) and overall survival were evaluated. CIK cell cytotoxicity against tumor cells was detected using a lactate dehydrogenase-based method. Serum cytokine levels in the immuno-CT group were detected using cytokine antibody arrays.

Results

The cytotoxicity of CIK cells was significantly enhanced by DC activation. The 2-year overall survival rate in the immuno-CT group was significantly improved compared with the CT group (94.7 ± 3.6% versus 78.8 ± 7.0%, P < 0.05). The 2-year DFS of these two groups showed no significant difference. DC-activated CIK cell treatment increased production of cytokines that have known anti-tumor effects, including IFN-γ, MIG, TNF-α and TNF-β, in patients who had no progression, but they were not found in patients who developed recurrence/metastasis.

Conclusions

This study suggests that the role of DC-activated CIK cells in improvement of chemotherapy for malignant tumor treatment is associated with up-regulation of the production of cytokines involved in the anti-tumor effect.

Introduction

Lung cancer is a leading cause of cancer-related death in both men and women world-wide. Surgical resection is a standard treatment for patients with early stages of lung cancer. However, it has been reported that tumors relapse in more than 50% patients after complete resection [1,2]. Thus post-operative chemotherapy is a required treatment for patients with lung cancer. Despite current advances in chemotherapy, the prognosis for patients with lung cancer is poor [3,4]. One limitation of current chemotherapy protocols is the lack of clinical evidence of an increase in patients' survival time. The largest meta-analysis published to date has shown that chemotherapy only increases the 1-year survival rate by 10% and median survival time by 6 weeks, compared with no chemotherapy treatment [5]. Therefore the development of effective therapies is clearly needed for treatment of lung cancer.

Recently, a number of cancer immunotherapy approaches for lung cancer have been developed and tested [6]. Increasing evidence suggests that adoptive immunotherapy reduces recurrence and metastasis rates of malignant tumors [7]. As a currently emerging adoptive immunotherapy, application of cytokine-induced killer (CIK) cells has shown significant anti-tumor activity in both clinical trials and animal studies, with cytotoxic effects against several tumor cells, including Hodgkin's disease, non-Hodgkin's lymphoma and hepatoma [1,8].

CIK cells are a unique population of cytotoxic T lymphocytes with a characteristic CD3⁺ CD56⁺ phenotype; they can be generated from cytokine cocktail-induced peripheral blood mononuclear cells (PBMC). These cells have shown higher proliferative and cytolytic activities compared with CD3⁻ CD56⁺ lymphokine-activated killer cells [2], for example CIK cells are cytolytic against chronic myeloid leukemia cells in vitro and in the SCID mouse tumor model. Interestingly, the anti-tumor activity of CIK cells can be enhanced by incubation with dendritic cells (DC), which are the most potent antigen (Ag)-presenting cells [5].

The purpose of this study was to evaluate the clinical efficacy of DC-activated CIK cell treatment following regular chemotherapy and the effects of this therapy on immune responses in patients with non-small cell lung cancer (NSCLC) after surgery. Our study shows that chemotherapy with DC-activated CIK cells significantly improves the overall survival rate and increases the production of cytokines that mediate anti-tumor effects. These findings suggest that DC-activated CIK cells may enhance the clinical efficacy of regular chemotherapy for malignant tumor treatment.

Section snippets

Patients

We performed a paired study to compare the clinical outcomes of patients with stage I–IIIa NSCLC after surgery receiving chemotherapy combined with adoptive immunotherapy containing natural killer (NK)-activated CIK cells (immuno-CT) or chemotherapy (CT) alone. The criteria for patient selection included being aged between 18 and 80 years, and having an expected survival duration of longer than 3 months, a Karnofsky performance status score higher than 40%, a white blood cell count higher than

Development of DC-activated CIK cells

To determine the immunologic effects of DC on activation of CIK cells, the phenotype and cytotoxicity of CIK cells before and after induction by DC were analyzed using flow cytometry and LDH, respectively. The population of CD3⁺ CD56⁺ CIK cells (19.02 ± 5.37%), which present higher proliferative and cytolytic activities compared with CD3⁻ CD56⁺ lymphokine-activated killer cells, was significantly increased by DC activation compared with CIK cells without DC activation (14.72 ± 5.03%, P < 0.05).

Discussion

Increasing clinical studies suggest that immunotherapy may serve as a strategy to improve cancer treatment. Adoptive immunotherapy using CIK cells has shown significant anti-tumor activity in pre-clinical experiments and animal tumor models [1,2,8]. Our study shows that Ag-loaded DC enhance the cytotoxicity of CIK cells against tumor cells. DC increased the population of the hallmark effector CD3⁺ CD56⁺ cells and decreased the proportion of CD4⁺ CD25⁺ cells that had potent immune suppressive

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Immune checkpoint inhibitor-based immunotherapy has revolutionized the treatment approach of non-small cell lung cancer (NSCLC). Monoclonal antibodies against programmed cell death-1 (PD-1) and PD-ligand 1 (PD-L1) are widely used in clinical practice, but other antibodies that can circumvent innate and acquired resistance are bound to undergo preclinical and clinical studies. However, tumor cells can develop and facilitate the tolerogenic nature of the tumor microenvironment (TME), resulting in tumor progression. Therefore, the immune escape mechanisms exploited by growing lung cancer involve a fine interplay between all actors in the TME. A better understanding of the molecular biology of lung cancer and the cellular/molecular mechanisms involved in the crosstalk between lung cancer cells and immune cells in the TME could identify novel therapeutic weapons in the old war against lung cancer. This article discusses the role of TME in the progression of lung cancer and pinpoints possible advances and challenges of immunotherapy for NSCLC.
CIK as therapeutic agents against tumors
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Cytokine Induced Killer (CIK) cells are ex vivo expanded and activated T lymphocytes obtained by sequential incubation of Peripheral Blood Mononuclear cells (PBMNC) with Interferon γ (IFNG), anti CD3 monoclonal antibody OKT3 and IL2.
These cells, while retaining few characteristics of the Effector memory T cells subpopulation, acquired during culture CD56 expression, as well as non specific, Natural Killer like, anti tumoral cytotoxicity.
CIK cells from human are equivalent to expanded NKT cells in mouse.
More interestingly, CIK cells show a potent intratumoral homing in several experimental models, followed by anti tumoral clinical activity in mice and humans.
In spite of extensive in vivo permanence and proliferation, CIK cells do not show cytotoxicity against normal targets and, particularly important, do not show Graft versus host disease when tested in allogeneic combinations (donor versus host) even in the haploidentical matching.
For the easiness of the laboratory preparations, the availability of clinical grade reagents, the production of Good Manufacturing Practice compliant methods, CIK cells have been extensively used for the treatment of cancer patients, in both hematologic and solid tumors, in both autologous and allogeneic combinations.
Several clinical protocol will be here discussed and summarised to show the feasibility of these passive transfer approaches, and also their very limited toxicity. Finally, preliminary indications on clinical efficacy, particularly in hematologic malignancies and against minimal residual disease, will be shown and discussed, as well as the future perspectives to optimize this adoptive passive cell immunotherapy strategy by gene transfer technology or bispecific monoclonal antibodies addition.
Influence of the number and interval of treatment cycles on cytokine-induced killer cells and their adjuvant therapeutic effects in advanced non-small-cell lung cancer (NSCLC)
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Cytokine-induced killer (CIK) cells have important therapeutic effects in adoptive cell transfer (ACT) for the treatment of various malignancies. In this study, we focused on in vitro expansion of CIK cells and their clinical efficacy in combination with chemotherapy in patients with advanced non-small-cell lung cancer (NSCLC).
A total of 64 patients with NSCLC (enrolled from 2011 to 2012), including 32 patients who received chemotherapy alone or with sequential radiotherapy (conventional treatment, control group) and 32 patients who received conventional treatment and sequential CIK infusion (study group), were retrospectively analyzed. The time to progression (TTP), overall survival (OS) and adverse effects were analyzed and the phenotype of lymphocytes in CIK population was also determined by flow cytometry.
After in vitro expansion, the average percentage of CIK cells was 26.35%. During the 54-month follow up, the median OS and TTP were significantly longer in the study group than in the control group (P = 0.0189 and P = 0.0129, respectively). The median OS of the ACT ≥ 4 cycles subgroup was significantly longer than that of the ACT < 4 cycles subgroup (P = 0.0316). The percentage of CIK cells in patients who received ≥ 4 cycles of ACT was higher than that in patients treated with < 4 cycles of ACT (P = 0.0376). Notably, CIK cells were difficult to expand in vitro in some patients after the first ACT cycle but became much easier as the treatment cycles increased monthly. Longer treatment interval negatively impacted the expansion of CIK cells.
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^*: These authors contributed equally

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Dendritic cell-activated cytokine-induced killer cells enhance the anti-tumor effect of chemotherapy on non-small cell lung cancer in patients after surgery

Background aims

Methods

Results

Conclusions

Introduction

Section snippets

Patients

Development of DC-activated CIK cells

Discussion

Semin Radiat Oncol

Clin Lung Cancer

Blood

Biol Blood Marrow Transplant

J Biol Chem

Unexpected long-term survival after low-dose palliative radiotherapy for non-small cell lung cancer

Cancer

CIK cells from patients with HCC possess strong cytotoxicity to multidrug-resistant cell line Bel-7402/R.World

Gastroenterol

Th1 bias in PBMC induced by multicycles of auto-CIKs infusion in malignant solid tumor patients

Cancer Biotherapy & Radiopharmaceuticals

Interactions between dendritic cells and cytokine-induced killer cells lead to an activation of both populations

J Immunother

Regulatory T cells and tumour immunity - observations in mice and men

Immunology