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Expansion, Purification, and Functional Assessment of Human Peripheral Blood NK Cells
Published on: February 3, 2011
Quantitative analysis of the effect of CD16 ligation on human NK cell proliferation
1Cancer Research Unit, Canberra Hospital, Australia.
Insights
CD16 ligation on natural killer (NK) cells stimulates proliferation but can cause progenitor cell loss. However, costimulation via melanoma cells enhances NK cell division without progenitor loss, indicating CD16
Area of Science:
- Immunology
- Cell Biology
Background:
- CD16 (Fc gammaRIIIA) is a low-affinity IgG receptor found on most human peripheral blood NK cells.
- CD16 ligation typically activates NK cell cytotoxicity, cytokine secretion, and apoptosis.
- The dual role of CD16 in NK cell activation and death requires further investigation.
Purpose of the Study:
- To quantitatively assess the impact of CD16 ligation on NK cell division and survival.
- To investigate how cellular costimulation influences CD16-mediated NK cell responses.
Main Methods:
- Human peripheral blood NK cells were labeled with carboxyfluorescein diacetate succinimidyl ester for quantitative tracking.
- NK cells were cultured with recombinant IL-2 (rIL-2) and subjected to CD16 ligation.
- NK cell proliferation and progenitor cell survival were analyzed under different stimulation conditions, including coculture with gamma-irradiated MM-170 melanoma cells.
Main Results:
- CD16 ligation in the presence of rIL-2 stimulated NK cell division but also led to significant NK progenitor cell loss.
- When NK cell proliferation was induced by coculture with MM-170 cells and rIL-2, CD16 ligation enhanced cell division.
- In the context of MM-170 cell coculture, CD16 ligation did not result in NK progenitor cell loss, and in some cases, was essential for proliferation.
Conclusions:
- CD16 functions as an activation receptor promoting NK cell proliferation.
- Cellular costimulation appears to modulate the balance between CD16-induced NK cell death and proliferation.
- These findings highlight the context-dependent effects of CD16 signaling on NK cell fate.
Abstract:
CD16 (Fc gammaRIIIA), the low affinity receptor for IgG, is expressed on the majority of human peripheral blood NK cells. Ligation of CD16 with mAb or immune complexes activates NK cell cytotoxicity and cytokine secretion, and stimulates death of activated NK cells by apoptosis. This study uses NK cells labeled with the stable intracytoplasmic fluorescent dye 5- and 6-carboxyfluorescein diacetate succinimidyl ester to provide quantitative data on the effect of CD16 ligation on NK cell division and NK cell survival. When NK cells are cultured with rIL-2 and CD16 is ligated, NK cell division is stimulated, but there also is a substantial loss of NK progenitor cells. When NK cell proliferation is stimulated by coculture with gamma-irradiated MM-170 malignant melanoma cells and rIL-2, CD16 ligation enhances entry of NK cells into division. In some cases, CD16 ligation is essential for NK cell proliferation stimulated by MM-170 cells. In these cultures, there is no loss of NK progenitor cells. This study demonstrates that CD16 is an activation receptor for NK cell proliferation, and suggests that cellular costimulation alters the balance between NK cell death and NK cell proliferation stimulated by CD16 ligation.
