Tuberc Respir Dis.  2010 Dec;69(6):456-464.

The Effect of Gefitinib on Immune Response of Human Peripheral Blood Monocyte-Derived Dendritic Cells

• Cho JH
• Kim MH
• Lee KH
• Kim KU
• Jeon DS
• Park HK
• Kim YS
• Lee MK
• Park SK

Abstract

BACKGROUND
Synergistic antitumor effects of the combined chemoimmunotherapy based on dendritic cells have been reported recently. The aim of this study is to search new applicability of gefitinib into the combination treatment through the confirmation of gefitinib effects on the monocyte derived dendritic cells (moDCs); most potent antigen presenting cell (APC).
METHODS
Immature and mature monocyte-derived dendritic cell (im, mMoDC)s were generated from peripheral blood monocyte (PBMC) in Opti-MEM culture medium supplemented with IL-4, GM-CSF and cocktail, consisting of TNF-alpha (10 ng/mL), IL-1beta (10 ng/mL), IL-6 (1,000 U/mL) and PGE2 (1 micro/mL). Various concentrations of gefitinib also added on day 6 to see the influence on immature and mature MoDCs. Immunophenotyping of DCs under the gefitinib was performed by using monoclonal antibodies (CD14, CD80, CD83, CD86, HLA-ABC, HLA-DR). Supernatant IL-12 production and apoptosis of DCs was evaluated. And MLR assay with [3H]-thymidine uptake assay was done.
RESULTS
Expression of CD83, MHC I were decreased in mMoDCs and MHC I was decreased in imMoDCs under gefitinib. IL-12 production from mMoDCs was decreased under 10 microM of gefitinib sinificantly. Differences of T cell proliferation capacity were not observed in each concentration of geftinib.
CONCLUSION
In spite of decreased expressions of some dendritic cell surface molecules and IL-12 production under 10 microM of gefitinib, significant negative influences of gefitinib in antigen presenting capacity and T cell stimulation were not observed.

Keyword

Dendritic cells; gefitinib; Interleukin-12; Apoptosis

MeSH Terms

Antibodies, Monoclonal
Apoptosis
Cell Proliferation
Dendritic Cells
Dinoprostone
Granulocyte-Macrophage Colony-Stimulating Factor
Humans
Immunophenotyping
Interleukin-12
Interleukin-4
Interleukin-6
Monocytes
Quinazolines
Tumor Necrosis Factor-alpha
Antibodies, Monoclonal
Dinoprostone
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-12
Interleukin-4
Interleukin-6
Quinazolines
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 Expression of cell surface markers before and after gefitinib treatment. CD14, CD80, CD83, CD86, MHC I and MHC II of mature monocyte-derived dendritic cells (A) and immature monocyte-derived dendritic cells (B). Grey histogram indicates the negative control staining and the number in each box represents mean fluorescence intensity (MFI).

• 

  Figure 2 2. IL-12 in supernatant of dendritic cells under various circumstance. IL-12 secretion of mature monocyte-derived dendritic cells shows decreasing tendency as increasing concentration of gefitnib. *p=0.036.

• 

  Figure 3 Apoptosis of monocyte-derived dendritic cells under the various concentration of gefitinib. The right two quadrants represent the apoptotic non-viable cells, positive for FITC-Annexin V binding. C: cocktail.

• 

  Figure 4 Mixed lymphocyte reaction assay. Irradiated DCs (104 cells/well) are cocultured with nylon wool-passed T cell (105 cells/well) ie, S : R=1 : 10. [3H]-thymidine uptake is represented as counts per minute (CPM) with seven wells from each seven donors. *p=0.924, †p=0.684.

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  Figure 5 Mixed lymphocyte reaction assay. The most striking increase of [3H]-thymidine uptake is shown in the ratio dendritic cell : T cell=1 : 10. S : R ratio rather than maturation status of dendritic cells exerts more profound influence on [3H]-thymidine uptake.

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