Immune Netw.  2012 Dec;12(6):269-276. 10.4110/in.2012.12.6.269.

Dendritic Cell (DC) Vaccine in Mouse Lung Cancer Minimal Residual Model; Comparison of Monocyte-derived DC vs. Hematopoietic Stem Cell Derived-DC

• Baek S
• Lee SJ
• Kim MJ
• Lee H

• KMID: 2150759
• DOI: http://doi.org/10.4110/in.2012.12.6.269

Abstract

The anti-tumor effect of monocyte-derived DC (MoDC) vaccine was studied in lung cancer model with feasible but weak Ag-specific immune response and incomplete blocking of tumor growth. To overcome this limitation, the hematopoietic stem cell-derived DC (SDC) was cultured and the anti-tumor effect of MoDC & SDC was compared in mouse lung cancer minimal residual model (MRD). Therapeutic DCs were cultured from either CD34+ hematopoietic stem cells with GM-CSF, SCF and IL-4 for 14 days (SDC) or monocytes with GM-CSF and IL-4 for 7 days (MoDC). DCs were injected twice by one week interval into the peritoneum of mice that are inoculated with Lewis Lung Carcinoma cells (LLC) one day before the DC injection. Anti-tumor responses and the immune modulation were observed 3 weeks after the final DC injection. CD11c expression, IL-12 and TGF-beta secretion were higher in SDC but CCR7 expression, IFN-gamma and IL-10 secretion were higher in MoDC. The proportion of CD11c+CD8a+ cells was similar in both DC cultures. Although both DC reduced the tumor burden, histological anti-tumor effect and the frequencies of IFN-gamma secreting CD8+ T cells were higher in SDC treated group than in MoDC. Conclusively, although both MoDC and SDC can induce the anti-tumor immunity, SDC may be better module as anti-tumor vaccine than MoDC in mouse lung cancer.

Keyword

Dendritic cell; Lung cancer; Anti-tumor immunity

MeSH Terms

Animals
Carcinoma, Lewis Lung
Dendritic Cells
Granulocyte-Macrophage Colony-Stimulating Factor
Hematopoietic Stem Cells
Interleukin-10
Interleukin-12
Interleukin-4
Lung
Lung Neoplasms
Mice
Monocytes
Peritoneum
T-Lymphocytes
Transforming Growth Factor beta
Tumor Burden
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-10
Interleukin-12
Interleukin-4
Transforming Growth Factor beta

Figure

• 

  Figure 1 Treatment schedule. D-0: LLC cells were inoculated 1×105/mouse i.v. D-1 and 8: In order to minimal residual model, therapeutic-DCs were injected twice by one week interval into the peritoneum of tumor cell inoculated-mice. D-29: Mouse sacrifice and immune monitoring. Each experimental group was composed of 5~7 mice.

• 

  Figure 2 Characterization of cultured DCs. DCs cultured from either CD34+ cells with GM-CSF, SCF and IL-4 for 14 days (SDC) or antibody-panned monocytes with GM-CSF and IL-4 for 7 days (MoDC). Each DC type was pulsed with LLC lysate (SDC/lysate and MoDC/lysate). Cytokines IL-12, IL-10, IFN-γ and TGF-β produced into the culture supernatant were measured by ELISA and expressed as pg/ml/1×106 cells. Statistical significance (p<0.05) was compared by asterisks between the SDC and MoDC. Statistical differences (p<0.05) between the lysate pulsed DCs were expressed by + signs.

• 

  Figure 3 Immunogenic phenotypes of cultured DCs were analyzed by flow cytometry. Dendritic cell marker CD11c as well as MHC II, CD8a and CCR7 with CD54 were measured to characterize and differentiate the cultured SDC and MoDC.

• 

  Figure 4 Anti-tumor effect of DCs for LLC formed in lung. Tumor bearing mice were sacrificed after three weeks from the last DC injection. (A) Circle represents number of tumor size over 0.2 mm in a mouse lung. Number and red line show mean value±standard error (SE). Same value represents one circle. Each experimental group was composed of 5~7 mice. (B) Pulmonary tissues of tumor-bearing mice were H&E stained.

• 

  Figure 5 Induction of tumor antigen-specific IFN-secreting cells. Tumor antigen-specific alterations in immunological parameters were analyzed with splenic lymphocytes obtained from the tumor-bearing mice treated with DC vaccine. As an effector molecule of therapeutic response, the proportion of IFN-secreting CD8+ T cells was observed by ELISPOT assay. Asterisk indicate the statistical significance (p<0.05) compared with MoDC.

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