Cancer Immunology, Immunotherapy

, Volume 61, Issue 10, pp 1683–1697 | Cite as

Tumor-associated myeloid cells can be activated in vitro and in vivo to mediate antitumor effects

  • Alexander L. Rakhmilevich
  • Mark J. Baldeshwiler
  • Tyler J. Van De Voort
  • Mildred A. R. Felder
  • Richard K. Yang
  • Nicholas A. Kalogriopoulos
  • David S. Koslov
  • Nico Van Rooijen
  • Paul M. Sondel
Original article

Abstract

Tumor growth is often accompanied by the accumulation of myeloid cells in the tumors and lymphoid organs. These cells can suppress T cell immunity, thereby posing an obstacle to T cell-targeted cancer immunotherapy. In this study, we tested the possibility of activating tumor-associated myeloid cells to mediate antitumor effects. Using the peritoneal model of B16 melanoma, we show that peritoneal cells (PEC) in tumor-bearing mice (TBM) had reduced ability to secrete nitric oxide (NO) following in vitro stimulation with interferon gamma and lipopolysaccharide, as compared to PEC from control mice. This reduced function of PEC was accompanied by the influx of CD11b+ Gr-1+ myeloid cells to the peritoneal cavity. Nonadherent PEC were responsible for most of the NO production in TBM, whereas in naïve mice NO was mainly secreted by adherent CD11b+ F4/80+ macrophages. Sorted CD11b+ Gr-1 monocytic and CD11b+ Gr-1+ granulocytic PEC from TBM had a reduced ability to secrete NO following in vitro stimulation (compared to naïve PEC), but effectively suppressed proliferation of tumor cells in vitro. In vivo, treatment of mice bearing established peritoneal B16 tumors with anti-CD40 and CpG resulted in activation of tumor-associated PEC, reduction in local tumor burden and prolongation of mouse survival. Inhibition of NO did not abrogate the antitumor effects of stimulated myeloid cells. Taken together, the results indicate that in tumor-bearing hosts, tumor-associated myeloid cells can be activated to mediate antitumor effects.

Keywords

Myeloid cells Anti-CD40 CpG Immunotherapy 

Abbreviations

IFN-γ

Interferon gamma

Macrophages

PEC

Peritoneal cells

TLR

Toll-like receptor

TAM

Tumor-associated macrophages

TNFα

Tumor necrosis factor alpha

TBM

Tumor-bearing mice

Supplementary material

262_2012_1236_MOESM1_ESM.doc (163 kb)
Supplemental figure. Effect of B16 cells in vivo and in vitro on PEC function and phenotype. A. C57BL/6 mice were injected i.p. with 1.6x107 B16 cells or left untreated. PEC were obtained 4 days later, placed in 96-well plate, and plastic-adherent cells were stimulated with IFN-γ (10 U/ml) and LPS (1 ng/ml). The supernatants were collected 48 hr later and NO activity was determined by nitrite levels. The results are presented as the Mean ± SEM nitrite concentration from 4 mice per group. B. PEC were obtained from naïve C57BL/6 mice, and 2x105 PEC were allowed to adhere to plastic for 2 hr followed by removal of nonadherent cells before adding B16 cells at indicated doses to adherent PEC in the presence of IFN-γ and LPS. NO activity in supernatants was determined 48 hr later. The results are presented as the Mean ± SEM nitrite concentration from triplicates. C. PEC obtained from naïve C57BL/6 mice (5x106) and RAW 247.7 macrophage cell line cells (2.5x106) were placed in the wells of the 6-well plates. PEC were allowed to adhere to plastic for 2 hr followed by removal of nonadherent cells. The cells were incubated in medium with or without gamma-irradiated (200 Gy) B16 cells (2.5x106) for 3 days, after which the cells were analyzed for co-expression of F4/80 and Gr-1 by flow cytometry. The representative dot plots are shown. (DOC 163 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Alexander L. Rakhmilevich
    • 1
    • 2
  • Mark J. Baldeshwiler
    • 1
  • Tyler J. Van De Voort
    • 1
  • Mildred A. R. Felder
    • 3
  • Richard K. Yang
    • 1
  • Nicholas A. Kalogriopoulos
    • 1
  • David S. Koslov
    • 1
  • Nico Van Rooijen
    • 5
  • Paul M. Sondel
    • 1
    • 2
    • 4
  1. 1.Departments of Human OncologyUniversity of WisconsinMadisonUSA
  2. 2.Paul P. Carbone Comprehensive Cancer CenterUniversity of WisconsinMadisonUSA
  3. 3.Department of Obstetrics and GynecologyUniversity of WisconsinMadisonUSA
  4. 4.Department of PediatricsUniversity of WisconsinMadisonUSA
  5. 5.Department of Molecular Cell BiologyVrije Universiteit Medical CentreAmsterdamThe Netherlands

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