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A Cell Free Assay System Estimating the Neutralizing Capacity of GM-CSF Antibody using Recombinant Soluble GM-CSF Receptor
Published on: June 27, 2011
The effects of recombinant human granulocyte-macrophage colony-stimulating factor on the induction of
S Okamura1, T Tanaka, S Yamaga
1Cancer Center, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Insights
Adding granulocyte-macrophage colony-stimulating factor (GM-CSF) to interleukin 2 (IL-2) cultures significantly increases lymphokine-activated killer (LAK) cell numbers. This enhances the generation of effector cells with potent cytotoxic activity against tumor cells.
Area of Science:
- Immunology
- Cell Biology
- Cancer Research
Background:
- Lymphokine-activated killer (LAK) cells are crucial for adoptive immunotherapy.
- Interleukin 2 (IL-2) is a key cytokine for LAK cell induction.
- Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine with known immunomodulatory effects.
Purpose of the Study:
- To investigate the effect of GM-CSF on LAK cell induction in vitro.
- To determine if GM-CSF enhances the number and cytotoxic activity of LAK cells generated with IL-2.
Main Methods:
- Human peripheral blood mononuclear cells were cultured with IL-2 and varying concentrations of GM-CSF.
- Viable cell counts were assessed using the trypan blue dye exclusion test.
- Cytotoxic effects against NK-resistant (Daudi) and NK-sensitive (K562) tumor cells were measured via 51Cr release assay.
Main Results:
- GM-CSF alone did not affect cell cytotoxicity or surface phenotype.
- GM-CSF significantly increased viable cell counts when combined with IL-2 (P < 0.01).
- The combination of GM-CSF and IL-2 led to a greater number of effector cells with substantial cytotoxic activity.
Conclusions:
- GM-CSF in combination with IL-2 enhances LAK cell expansion.
- This combination is beneficial for generating a larger pool of potent LAK effector cells for potential therapeutic applications.
- GM-CSF plays a supportive role in IL-2-mediated LAK cell induction.
Abstract:
Granulocyte-macrophage colony-stimulating factor (GM-CSF) was added to a culture of human peripheral blood mononuclear cells in the presence of interleukin 2 (IL-2) in vitro to elucidate its effect on the induction of lymphokine-activated killer (LAK) cells. Viable cell counts of cultured cells and their cytotoxic effects against natural killer (NK) cell-resistant Daudi cells and NK cell-sensitive K562 cells were measured using the trypan blue dye exclusion test and a 51Cr release assay from the tumor cells, respectively. Although GM-CSF alone did not influence either the cytotoxicities or the surface phenotypes of the cultured cells, the viable cell counts were significantly increased by the addition of GM-CSF in the presence of IL-2 (P less than 0.01). These findings indicate that the addition of GM-CSF in the presence of IL-2 during the induction of LAK cells is useful for obtaining a larger number of effector cells which possess substantial cytotoxic activity.
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