Elsevier

Vaccine

Volume 27, Issue 44, 19 October 2009, Pages 6210-6216
Vaccine

Improved therapeutic efficacy using vaccination with glioma lysate-pulsed dendritic cells combined with IP-10 in murine glioma

https://doi.org/10.1016/j.vaccine.2009.08.002Get rights and content

Abstract

The purpose of the present study was to evaluate the therapeutic efficacy of glioma lysate-pulsed DCs in combination with plasmid DNA vector encoding the murine interferon-induced protein of 10 kDa (IP-10 or CXCL10) gene. Mouse models of brain glioma (GL261) were treated with combining glioma lysate-pulsed DCs with direct intratumoral injection of a nonviral plasmid DNA vector encoding the murine IP-10 gene. The survival of mice bearing GL261 glioma was observed. Enzyme-linked immuno-spot assay was used to determine the frequency of brain-infiltrating lymphocytes (BILs) capable of responding to GL261. Cytolytic T lymphocyte (CTL) response was measured by cytotoxic assay, vessel density and tumor cell proliferation were observed by immunostaining, and tumor apoptosis was determined by TUNEL staining. The results revealed that the combination therapy groups showed more significantly enhanced anti-tumor activity, attraction of lymphocytes into tumor tissues, apoptosis of tumor cells, and reduced neovascularization, cell proliferation, and developed a strong CTL response in these mice. In summary, the therapy of glioma lysate-pulsed DCs combined with the IP-10 gene has significant synergistic effect against glioma.

Introduction

Malignant gliomas are the most common primary brain tumor of the central nervous system in adults [1]. The prognosis of malignant glioma is usually poor, with life expectancy <1 year from time of diagnosis and a 5-year survival rate of <5.5% [1], [2], [3], [4], [5]. Complete surgical resection remains the primary mode of therapy for localized lesions, whereas numerous adjuvant modalities have been investigated for benefit in metastatic disease. So far, mainly the immune-based modalities, such as dendritic cells (DCs)-based vaccine, cytotoxic T lymphocyte (CTL), lymphokine-activated killer cells (LAK), natural killer cells (NK) and cytokines, still hold some promise. However, these strategies require further study to improve consistent tumor destruction, extended life span, safety and feasibility for cancer patients.

DCs are unique highly potent antigen-presenting cells capable of sensitizing CD4+ T cells and CD8+ T cells [6]. With the availability of isolation and bulk propagation of DCs in vitro, many methods for antigen priming aimed at inducing anti-tumor immune responses by DC-based vaccination have been attempted. For example, the DCs were either transduced by viral vectors encoding the tumor antigens [7] or pulsed with a tumor cell lysate [8], [9], apoptotic tumor cells [10], synthetic peptide [11], [12], tumor RNA [13] and DNA [14], [15]. Although anti-tumor cellular immune responses could be induced by DC vaccination, clinical objective responses were limited. Thus, a new strategy for DC-based tumor vaccines is expected to improve the clinical effectiveness of this treatment.

Interferon (IFN)-γ-inducible protein-10 (IP-10) [16] belongs to the CXC chemokine family known to stimulate the IP-10 receptor CXCR3 [17]. IP-10 binds to a seven-transmembrane G protein-coupled receptor, CXCR3, expressed on activated T cells, leading to chemotaxis [17]. Recently Nishimura et al reported that IP-10 plays a critical role in the homing of Tc1 into central nervous system (CNS) tumors [18]. In addition, it can also inhibit tumoral angiogenesis [19], [20]. Therefore, IP-10 has been involved in the anti-tumor immune responses by recruitment of T cells to the malignancies and the nonimmune responses by its anti-angiogenesis effect.

In the present study, we have demonstrated that IP-10 can potentiate the anti-tumor effects of glioma lysate-pulsed DCs in vivo during treatment of mouse glioma. This study suggests that immunization of tumor lysate-pulsed DCs in combination with IP-10 may be useful for suppressing the growth of recurring tumors after primary therapy of human glioma.

Section snippets

Plasmids

Total RNA was extracted from IFN-γ (1000 U/ml)-stimulated L929 cell line using Trizol (Invitrogen, Carlsbad, CA). The oligonucleotides used for RT-PCR were as follows: IP-10 forward (5′-CGGAATTCATCAGCACCATGAACCCAAGT-3′), and backward (5′-GCGTGGCTTCTCTCCAGTT-3′), and GAPDH forward (5′-CTGCACCACCAACTGCTTAG-3′) and backward (5′-GTCTGGGARGGAAARRGRGA-3′). IP-10 cDNA was amplified with RT-PCR and subsequently subcloned into the EcoRI and XhoI sites of the ampicillin-selectable mammalian expression

Anti-tumor efficacy of combination GL261 lysate-pulsed DCs and IP-10

We tested the effects of GL261 lysate-pulsed DCs (subcutaneously with 1 × 106 GL261 lysate-pulsed DCs on day 1, 8 and 15 after intracranial inoculation of GL261 cells) and IP-10 (three treatments in the same inoculation location on 5, 10 and 15 days after the i.c. tumor challenge), alone or together (subcutaneously with 1 × 106 GL261 lysate-pulsed DCs on day 1, 8 and 15 after intracranial inoculation of GL261 cells and three treatments in the same inoculation location on 5, 10 and 15 days after the

Discussion

The potential of DC-based vaccines to eradicate unresectable tumors has stimulated intense investigation. Most studies have focused on improving vaccine efficacy using adjunct treatment or the expression of cytokines and costimulatory molecules [25], [26], [27], [28], [29], [30], [31], [32], [33]. Little is known about the impact of IP-10 on the efficacy of the DC vaccine. Considering that IP-10 is a potent inhibitor of angiogenesis and plays a critical role in the homing of Tc1 into CNS tumors

Acknowledgments

This work was supported by the grant from the National Natural Science Foundation of China (No. 30672151) and the National Natural Science Foundation of China (No. 30801355).

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    These authors contributed equally to this work.

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