| |
Mean count of neutrophils attached to one melanoma cell by mechanism of frustrated phagocytosis
|
|---|
|
Ligand
|
20 min.
|
30 min.
|
40 min.
|
|---|
|
Mannan-BAM
|
0.333
|
0.556
|
0.733
|
|
Mannan free
|
0.111
|
0.000
|
0.000
|
|
f-MLFKK-BAM
|
0.589
|
0.311
|
0.360
|
|
f-MLF free
|
0.480
|
0.111
|
0.000
|
|
PBS (control)
|
0.000
|
0.000
|
0.000
|
- B16-F10 melanoma cells were incubated (30 min, 37 °C) with 0.02 mM mannan-BAM or 0.05 mM f-MLFKK-BAM in culture medium and then subsequently washed. Suspension of bone marrow neutrophils (90% purity) primed with GM-CSF + TNF-alpha (+ laminarin in case of mannan-BAM) in culture medium was added to B16-F10 cells. Free ligands were added in a concentration of 0.02 mM (mannan) and 0.05 mM (f-MLFKK). Neutrophils and melanoma cells were incubated at a 2:1 ratio
- The rate of frustrated phagocytosis events was estimated by light microscopy: Frustrated phagocytosis was defined as neutrophil/melanoma cell contact, where neutrophil adhere tightly to B16-F10 cell. Such contact is further characterized by neutrophil flattening and gaining of waning moon shape [5]
