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Updated: Aug 8, 2026
Flow Cytometric Analysis of Natural Killer Cell Lytic Activity in Human Whole Blood
Published on: March 17, 2017
Kinetic analysis of human IL-2 activated cytotoxic cells
A V LeFever1, V D Piaskowski, J T Casper
1Department of Pediatrics, Medical College of Wisconsin, MACC Fund Research Center, Milwaukee 53226.
Insights
Kinetic analysis quantifies lymphokine-activated killer (LAK) cell activity against tumor cells. Recombinant interleukin-2 (rIL-2) generated more effective LAK cells than lymphokine-conditioned medium (LCM).
Area of Science:
- Immunology
- Cellular Biology
- Cancer Research
Background:
- Peripheral blood mononuclear cells (PBMC) can be activated to become lymphokine-activated killer (LAK) cells.
- LAK cells are crucial in adoptive immunotherapy for cancer treatment.
- Understanding LAK cell lytic mechanisms is vital for optimizing cancer therapies.
Purpose of the Study:
- To define lytic events in LAK cells using kinetic analysis.
- To quantitatively assess LAK cell functional properties against tumor targets.
- To compare the efficacy of LAK cells activated by different methods.
Main Methods:
- Kinetic analysis was employed to determine Vmax and Km values.
- Lytic activity was assessed against NK-resistant and NK-sensitive tumor cell lines.
- LAK cells were generated using lymphokine-conditioned medium (LCM) and recombinant interleukin-2 (rIL-2).
Main Results:
- Kinetic parameters (Vmax, Km) were determined for LAK cell-mediated lysis.
- LAK cells activated by rIL-2 demonstrated higher lytic efficiency than those activated by LCM.
- T-depleted LAK cell populations exhibited the highest maximum rate of lysis (Vmax).
Conclusions:
- Kinetic analysis provides a quantitative method to evaluate LAK cell function.
- LAK cell generation using rIL-2 and T-depletion enhances lytic efficiency.
- This approach can assess the impact of various factors on LAK cell activity for therapeutic applications.
Abstract:
Kinetic analysis was used to define lytic events in peripheral blood mononuclear cells (PBMC) activated in lymphokine conditioned medium (LCM) and recombinant interleukin-2 (rIL-2). This analysis provided quantitative information on the functional properties of these lymphokine-activated killer (LAK) cells against NK-resistant and NK-sensitive tumor cell lines. The maximum rate of target cell lysis (Vmax) and Km (target cell number resulting in 1/2 Vmax) were determined. IL-2 activated effector cells that bound to target cells also lysed them (i.e., non-lytic bystander lymphocytes did not influence the determination of kinetic parameters) in contrast to lysis mediated by unactivated NK cells. The extent of LAK cell binding to tumor target cells was dependent upon the tumor type. LAK cell frequency determinations were calculated where Km approximated the concentration of LAK cells that were capable of killing a particular target. LAK cells generated in rIL-2 were lytically more efficient than those activated in LCM, and T-depletion resulted in a LAK population with the highest maximum rate of lysis. The use of kinetic analysis to evaluate LAK cell frequencies and quantitate lytic events will be useful in determining the effects of drugs, biological response modifiers and disease states on LAK cell function.
