Elsevier

Surgery

Volume 142, Issue 5, November 2007, Pages 749-760
Surgery

Original communication
Synergistic effect of intratumoral IL-12 and TNF-α microspheres: systemic anti-tumor immunity is mediated by both CD8+ CTL and NK cells

https://doi.org/10.1016/j.surg.2007.05.008Get rights and content

Neoadjuvant immunotherapy with the combination of intratumoral IL-12 and TNF-α encapsulated in poly-lactic acid microspheres (PLAM) generate a greater systemic immune response than either cytokine alone. We sought to examine the effector cells responsible for this synergy using the poorly immunogenic B16 melanoma and MCA205 sarcoma cell lines. Splenocytes from MCA205 bearing mice treated with IL-12 and TNF-α PLAM contained significantly more tumor-specific IFN-γ secreting cells than IL-12 alone. Adoptive transfer of lymphocytes from mice treated by the combination mediated significant tumor regression in mice bearing established pulmonary metastases. In mice bearing bilateral tumors, treatment of the primary with IL-12 and TNF-α PLAM, resulted in suppression of contralateral tumor growth. Both the local and distant effects were absent in mice depleted of CD8+ T-cells. In B16 bearing mice with established pulmonary disease, only the combination of intratumoral IL-12 and TNF-α resulted in a significant reduction of lung nodules. Both the local and distant effects were eradicated in mice depleted of either CD8+ T-cells or NK cells. The local and sustained release of IL-12 and TNF-α using PLAM synergistically activate both a cytotoxic T-cell and NK cell response, although their impact varies with MHC class I expression.

Section snippets

Animals and tumors

Six- to 8-week old female C57Bl6 mice were purchased from the Jackson Laboratory (Bar Harbor, NE) and maintained in specific pathogen-free conditions at the Animal Maintenance Facility of the University of Michigan Medical Center. MCA205 is a poorly immunogenic fibrosarcoma cell lines induced by 3-methylcholanthrene and syngeneic to C57Bl6 mice.7, 8 B16-BL6 (B16) is also a poorly immunogenic cell line; a melanoma of spontaneous origin that has been studied extensively.9 Cell lines were

Intratumoral injection of IL-12 and TNF-α microspheres confers systemic Anti-Tumor immunity

The synergy between IL-12 and TNF-α in MCA205 bearing C57Bl6 mice in generating a systemic anti-tumor response was confirmed via ELISPOT assay of the splenocytes to quantify tumor-specific IFN-γ producing cells (Fig 1). IL-12 alone did result in a significant increase in tumor specific T-cells compared to mice treated with BSA loaded microspheres or surgical excision. However, splenocytes from mice treated intralesionally with IL-12 and TNF-α demonstrated a dramatic increase in MCA205 specific

Discussion

Research in immunotherapy has focused on the post-operative setting, which is caught in a catch-22. The most immunogenic approaches have been those that utilize autologous tumor or tumor-infiltrating lymphocytes, but these approaches require an adequate amount of harvestable tumor, limiting their applicability to those patients with advanced disease. Approaches that are more clinically feasible in patients with only microscopic residual disease, such as peptide or allogeneic cellular vaccines,

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    Funded in part by NIH Grant CA102602-01 and a grant from the Gillson-Longenbaugh Foundation.

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