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Table 2 Interactions between neutrophils and melanoma cells with or without anchored ligands of phagocytic receptors

From: Innate immunity based cancer immunotherapy: B16-F10 murine melanoma model

 

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

  1. 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
  2. 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]