Inhibition of neutrophil apoptosis after severe trauma is NFkappabeta dependent.
BACKGROUND
Systemic inflammation may inhibit neutrophil (PMN) apoptosis and promote multiple organ dysfunction syndrome. We hypothesize that severe trauma causes dysregulation of PMN apoptosis.


METHODS
Neutrophils were isolated from trauma patients (24-72 hours after injury; n = 16) and controls (healthy volunteers) and incubated for 18 hours. In separate experiments, control cells were treated +/- the nuclear factor kappa beta (NFkappabeta) inhibitor pyrrolidinithiocarbamate then incubated with 25% patient or control plasma. Apoptosis was quantified by enzyme-linked immunosorbent assay for histone-associated DNA and annexin V fluorescence-activated cell sorter. NFkappabeta activation was determined by Western blot for phosphorylated I kappabeta.


RESULTS
Apoptosis was inhibited in trauma patient PMN. Neutrophil apoptosis correlated with multiple organ dysfunction syndrome score, Acute Physiology and Chronic Health Evaluation II, and platelet count. Patient plasma inhibited apoptosis and induced phosphorylation of I kappabeta in control cells. Inhibition of PMN apoptosis by patient plasma was blocked by pretreatment with pyrrolidinithiocarbamate.


CONCLUSIONS
NFkappabeta-dependent inhibition of neutrophil apoptosis occurs after trauma. Early inhibition of PMN apoptosis is dependent on the magnitude of injury.
Nolan
B
B
Department of Surgery, University of Massachusetts Medical School, Worcester 01655, USA.
Collette
H
H
Baker
S
S
Duffy
A
A
De
M
M
Miller
C
C
Bankey
P
P
D017209:Apoptosis / Q000502:physiology*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D006657:Histones / Q000032:analysis; D006801:Humans; D009102:Multiple Organ Failure / Q000097:blood; D016328:NF-kappa B / Q000032:analysis / Q000037:antagonists & inhibitors / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology*; D010766:Phosphorylation; D011759:Pyrrolidines / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014947:Wounds and Injuries / Q000097:blood*
D006657:Histones; D016328:NF-kappa B; D011759:Pyrrolidines; D013859:Thiocarbamates; C020972:pyrrolidine dithiocarbamic acid