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Expansion, Purification, and Functional Assessment of Human Peripheral Blood NK Cells
Published on: February 2, 2011
Glutamine requirements in the generation of lymphokine-activated killer cells
A Juretic1, G C Spagnoli, H Hörig
1Departments of Surgery and Research, University of Basel, ZLF 401, Hebelstrasse 20, CH-4031 Basel, Switzerland.
Insights
Glutamine (GLN) supports lymphokine-activated killer (LAK) cell generation. GLN deficiency limits effector cell numbers, impacting LAK cell killing activity but not broad-range killing capacity.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Lymphokine-activated killer (LAK) cells are crucial for cellular immunotherapy.
- Glutamine (GLN) is a key nutrient for immune cell function.
- Understanding GLN's role in LAK cell generation is vital for optimizing immunotherapy protocols.
Purpose of the Study:
- To investigate the role of glutamine (GLN) in the generation of lymphokine-activated killer (LAK) cell activity.
- To determine how different concentrations of GLN affect LAK cell proliferation and cytotoxic potential.
- To elucidate the impact of GLN on LAK cell generation using both bulk and limiting dilution culture systems.
Main Methods:
- Peripheral blood mononuclear cells (PBMC) from healthy donors were cultured with recombinant interleukin-2 (rlL-2).
- PBMC were cultured in the presence of varying concentrations of glutamine (GLN).
- Cytotoxic assays were performed using NK-sensitive (K562) and NK-resistant (1301, U-937) target cells. Limiting dilution (LD) culture was also employed.
Main Results:
- Glutamine (GLN) significantly affected the lytic potential of cultured cells.
- Cell proliferation with IL-2 stimulation showed significant differences in cultures with and without GLN.
- The frequency of responding cells did not significantly differ across varying GLN concentrations.
Conclusions:
- Glutamine (GLN) plays a supportive role in the generation of LAK cells.
- GLN deficiency limits the number of effector cells generated, thereby affecting LAK cell killing activity.
- The acquisition of broad-range killing capacity in LAK cells was not affected by GLN levels.
Abstract:
The role of glutamine (GLN) in the generation of lymphokine-activated killer (LAK) cell activity was investigated. LAK cells were derived from healthy donors and peripheral blood mononuclear cells (PBMC) were obtained using either unseparated PMBC or DR(-) CD3(-) CD16(+) CD56(+) enriched cells. PBMC were cultured for 6 or 10 days in medium supplemented with recombinant interleukin-2 (rlL-2; 100 U/ml) in the presence of different concentrations of GLN. K562 (natural killer-NK-sensitive targets), 1301 and U-937 (NK-resistant targets) cells were used as targets in the cytotoxic assays. Furthermore, the limiting dilution (LD) culture system was applied as an alternative to the bulk cell culture system. It was found that GLN affects the lytic potential of cultured cells while the frequency of responding cells did not significantly differ between the compared cell cultures performed in the presence of different amounts of GLN. Data on cell proliferation with IL-2 stimulation showed significant differences in cultures performed in the presence or absence of GLN. The results of present investigation suggest a supportive role of GLN in the generation of LAK cells. GLN deficit affects LAK cell killing activity by limiting the number of generated effector cells while acquisition of broad-range killing capacity was not affected.
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