Group Title: Critical Care
Title: Mobilization of leukocytes in patients with severe sepsis and septic shock is associated with increased apoptosis, as detected by Annexin V binding
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Title: Mobilization of leukocytes in patients with severe sepsis and septic shock is associated with increased apoptosis, as detected by Annexin V binding
Series Title: Critical Care
Physical Description: Archival
Creator: Weiss,M.
Barth,E.
Fischer,G.
Elsharkawi,M.
Moldawer,L. L.
Georgieff,M.
Schneider,E. M.
Publication Date: 2001
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General Note: Start pageP064
General Note: M3: 10.1186/cc1131
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Bibliographic ID: UF00100066
Volume ID: VID00001
Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: Open Access: http://www.biomedcentral.com/info/about/openaccess/
Resource Identifier: issn - 1364-8535
http://ccforum.com/content/5/S1/P064

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extracellular glucose concentrations. If this occurs in vivo, micro- References:
bial killing by neutrophils may be impaired in patients with acute 1. Zhang et al: J Biol Chem (in press). http://jbc.org/
hyperglycemia. 2. Marklund et al: Eur J Biochem 1974, 47:469.

P64 Mobilization of leukocytes in patients with severe sepsis and septic shock is associated with increased apoptosis, as
detected by Annexin V binding

M Weiss*, E Barth*, G Fischer*, M Elsharkawi*, LL Moldawert, M Georgieff*, EM Schneider*
*Department of Anesthesiology, University Hospital, Steinhoevelstr 9, D-89070 Ulm, Germany; *Department of Surgery, University of Florida
College of Medicine, Gainesville, FL 32610, USA


Objectives: To clarify whether mobilization of leukocytes during
human severe sepsis and septic shock occurs independently of
apoptosis.




Methods: Over a 6 month period, 33 patients with severe sepsis
or septic shock were studied in the intensive care unit. Annexin V
binding by leukocytes was determined daily using flow cytometry
and FITC-labeled Annexin V. Transient leukocytosis, or peaks in
leukocyte counts, were defined as individual increase of at least
30% within 2 days, followed by a decrease of at least 30% within
the following 2 days.


Results: Nine, 14 and 10 peaks in neutrophil, monocyte and lym-
phocyte counts, respectively, were observed in 6, 9 and 10
patients; in all of these patients, increased Annexin V binding by
neutrophils, monocytes and lymphocytes, respectively, by 69% up
to 809% (median 2150/0), by 32% up to 973% (median 3300/0),
and by 32% up to 4713% (median 2240/0), respectively, paralleled
the neutrophil, monocyte and lymphocyte (in 13/14 episodes)
peaks, respectively. During periods in which neutrophil, monocyte
and lymphocyte numbers were stable, Annexin V binding was con-
stant as well.

Conclusions: In conclusion, mobilization of leukocytes during
severe sepsis and septic shock in critically ill patients is associated
with increased apoptosis, as determined by Annexin V.


P65 C5a receptor expression on leukocytes from patients with severe sepsis or septic shock

M Furebring*, L Hakansson*, P Venge*, A Siegbahn*, B Nilssont, J Sj6olin*
*Department of Medical Sciences, and *Department of Oncology, Radiology and Clinical Immunology, University Hospital, 751 85 Uppsala,
Sweden


Background: Recently animal studies have been presented sug-
gesting that complement factor C5a blockade might be of benefit
in patients with severe sepsis/septic shock. In one study the
expression of the C5a receptor (CD88) on the granulocytes from
septic animals was increased. The aim of the present investigation
was to study the CD 88 expression on leukocytes in human sepsis.


Methods: Twelve ICU patients fulfilling the ACCP/SCCM criteria
for severe sepsis/septic shock were prospectively included into the
study as early as possible in their septic course. Blood samples for
analyses of leukocyte receptor expression and complement factors
were taken on day 1, 3 and 15. The leukocytes were isolated from
heparinised whole blood and labelled with CD88 antibodies. As
controls leukocytes from 20 healthy individuals were used. The
samples were analysed by the use of flow cytometry and results
presented as mean fluorescence intensity (MFI). The complement
proteins C3a and terminal complement complex (TCC) were
analysed in EDTA plasma by capture ELISA techniques. Levels of
TNF-a, IL-6, IL-8, IL-10, IL-1ra, and MCP-1 were analysed in EDTA
plasma with ELISA technique.


Results: On day 1 10/11 patients had increased levels of C3a,
1144 138 ng/ml, (mean SE) (normal range: 92-268) and
11/11 of TCC 146 46 AU/ml (normal range: 12-56). CD88
expression on the granulocytes in the control group was 63 4. In
comparison with the controls, the patients with severe
sepsis/septic shock had significantly lowered values: on day 1
37 5 (P< 0.001), on day 3 45 8 (P < 0.05), and on day 15
51 8 (P < 0.05). The granulocyte expression of CD88 on day 1
correlated negatively to APACHE-II score at inclusion (r = -0.59,
P< 0.05). Besides a weak correlation to IL-1 ra, there were no sig-
nificant correlations to the other cytokines. In the patient group, the
CD88 expression on the monocytes did not change during the
observation time and did not differ from that in the control group.

Conclusion: Although a transient increase at an earlier stage of
the septic course cannot be excluded, our results demonstrate that
the expression of the C5a receptor on granulocytes at the time
when diagnosis can clinically be made is low, despite an activa-
tion of the complement system. Our result suggest that C5a block-
ade in human sepsis might be of a more limited value than that
found in animal experiments.


P66 Terminal complement complex in porcine septic shock with substantial capillary leak syndrome

M Cobas-Meyer*, G Marxt, F Kube*, B Vangerow*, T Schuerholz*, J Schmidtko*, M Winkler*, KF Gratz, M Leuwert, H Rueckoldt*
*Department of Anaesthesiology, *Department of Viceral- and Transplantsurgery, and Department of Diagnostic Radiology, Hannover
Medical School, Carl-Neuberg-Str 1, D-30625 Hannover, Germany; *Department of Anaesthesia, Royal Liverpool University Hospitals,
Liverpool L69 3GA, UK


Introduction: In septic shock with capillary leak syndrome (CLS), it
has been suggested that hemodilution, and capillary leakage of
protein may account in part for reduced levels of complement pro-


teins observed in sepsis Aim of this study was to determine com-
plement activation by terminal complement complex (TCC) in
septic shock under the conditions of substantial CLS.




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