Isolation of human eosinophils: microbead method has no impact on IL-5 sustained viability.
OBJECTIVE
In a recent issue of Experimental Dermatology (18, 2009, 654), Schefzyk et al. concluded that multi-antibody eosinophil isolation (Miltenyi) should be abandoned, as differential purity was minimal, and eosinophils underwent accelerated apoptosis when compared with those isolated with traditional anti-CD16 microbeads. Our intent was to investigate the universality of these findings.


METHODS
We isolated eosinophils from normal donor granulocyte packs using two methods, and evaluated purity, viability and annexin-V/propidium-iodide staining.


RESULTS
Purity was substantially greater when multi-antibody isolation was used for eosinophil isolation from granulocyte packs (98% vs 69%). No differential survival was detected when eosinophils were maintained in culture with or without interleukin-5.


CONCLUSIONS
Multi-antibody eosinophil isolation represents a substantial advantage over anti-CD-16 microbeads when isolating large numbers of eosinophils from concentrated leucocyte preparations. No differential survival was observed. While appropriate consideration of methods is always crucial, multi-antibody eosinophil isolation should not be abandoned completely.
Percopo
Caroline M
CM
Dyer
Kimberly D
KD
Killoran
Kristin E
KE
Rosenberg
Helene F
HF
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology; D017209:Apoptosis / Q000187:drug effects; D019921:Biotinylation; D002452:Cell Count; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D004804:Eosinophils / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D015848:Interleukin-5 / Q000494:pharmacology*; D008863:Microspheres; D009000:Monocytes / Q000166:cytology; D009504:Neutrophils / Q000166:cytology
D000906:Antibodies; D015703:Antigens, CD; C508599:IL5 protein, human; D015848:Interleukin-5