1214 Crit Care Med 2012 Vol. 40, No. 4
C
erebral dysfunction is a com-
mon complication of sepsis,
and it has been shown to be as-
sociated with adverse outcomes
(1). Although the exact pathophysiol-
ogy remains poorly understood, cerebral
endothelial activation and the alteration
of the blood-brain barrier (BBB) have
been documented in in vivo and in vitro
models of the septic brain (2–4). However,
the morphologic changes in brain struc-
ture associated with sepsis, and the exact
cellular pathology underlying BBB dys-
function, have not been clearly identified
(5, 6).
Manipulating the inflammation caus-
ing endothelial activation and BBB leak-
age has been investigated as a therapeutic
target in sepsis-induced brain dysfunc-
tion. Different pharmacologic approaches
on lipopolysaccharide (LPS)-induced
changes in the BBB permeability have
previously been investigated in vitro (4).
Endothelial cells that had been pretreated
with dexamethasone were shown to be
more resistant to LPS-induced disruption
of the BBB (4). In another trial, LPS has
been targeted as a therapy option, and the
binding of circulating LPS by the serum
amyloid P component was shown to have
protective benefits in mice (7). Another
therapeutic option for the prevention
of sepsis-induced BBB breakdown was
demonstrated through the inhibition of
complement activation using an anti-C5a
antibody (8).
The targeting of complex immune ac-
tivation in neuroinflammation through
the use of immunoglobulin therapy for
immunomodulation has been used to
treat a variety of inflammatory and au-
toimmune diseases (9). Immunoglobulin
therapy has become an established treat-
ment for an array of neurologic dis-
orders, such as chronic inflammatory
demyelinating polyneuropathy, multi-
focal motor neuropathy, and Guillain-
Barré syndrome (10, 11). The effects of
Interventions: The effects of immunoglobulin G and immuno-
globulins enriched with immunoglobulin A and immunoglobulin M
on blood-brain barrier integrity and survival rates in septic rats
were comparatively investigated.
Measurements: Sepsis was induced by cecal ligation and per-
foration in Sprague-Dawley rats. The animals were divided into
the following groups: sham, cecal ligation and perforation, ce-
cal ligation and perforation plus immunoglobulin G (250 mg/kg,
intravenous), and cecal ligation and perforation plus immuno-
globulins enriched with immunoglobulin A and immunoglobulin M
(250 mg/kg, intravenous). Immunoglobulins were administered
5 mins before cecal ligation and perforation and the animals were
observed for behavioral changes for 24 hrs following cecal ligation
and perforation. Blood-brain barrier permeability was function-
ally and structurally evaluated by determining the extravasation
of Evans Blue and horseradish peroxidase tracers, respectively.
Immunohistochemistry and Western blotting for occludin were
performed.
Main Results: The high mortality rate (34%) noted in the sep-
tic rats was decreased to 15% and 3% by immunoglobulin G and
immunoglobulins enriched with immunoglobulin A and immuno-
globulin M, respectively (p , .01). Both immunoglobulin G and im-
munoglobulins enriched with immunoglobulin A and immunoglob-
ulin M alleviated the symptoms of sickness behavior in the septic
rats, with the animals becoming healthy and active. Increased
extravasation of Evans Blue into the brain tissue of the septic rats
was markedly decreased with the administration of both immuno-
globulin G and immunoglobulins enriched with immunoglobulin A
and immunoglobulin M (p , .01). Occludin expression remained
essentially unchanged in all groups, including the cecal ligation
and perforation group. In the cecal ligation and perforation group,
increased luminal and abluminal vesicles containing electron-
dense horseradish peroxidase-reaction product were noted in the
cytoplasm of endothelial cells located in the hippocampus and
the cerebral cortex. Tight junction was ultrastructurally intact,
suggesting that the transcellular pathway is responsible for the
blood-brain barrier breakdown in sepsis. Following immunoglob-
ulin G or immunoglobulins enriched with immunoglobulin A and
immunoglobulin M treatment, no ultrastructural evidence of leaky
capillaries in the brain was observed in the septic rats, indicat-
ing the blockade of the transcellular pathway by immunoglobulins
administration.
Conclusions: Our study suggests that immunoglobulin G
and immunoglobulins enriched with immunoglobulin A and
immunoglobulin M improve the integrity of the blood-brain
barrier and inhibits cecal ligation and perforation-induced
symptoms of sickness behavior in rats. (Crit Care Med 2012;
40:1214–1220)
KEY WORDS: blood-brain barrier; immunoglobulin A; immuno-
globulin G; immunoglobulin M; sepsis
Laboratory Investigations
Intravenous immunoglobulins prevent the breakdown of the blood-
brain barrier in experimentally induced sepsis*
Figen Esen, MD; Evren Senturk, MD; Perihan E. Ozcan, MD; Bulent Ahishali, MD; Nadir Arican, MD;
Nurcan Orhan; Oguzhan Ekizoglu, MD; Mutlu Kucuk, PhD; Mehmet Kaya, PhD
*See also p. 1368.
From the Departments of Anesthesiology (FE, ES,
PEO), Histology and Embryology (BA), Forensic Medicine
(NA), and Physiology (MK), Institute of Experimental
Medicine (NO, MK), Istanbul University, and Dr. SadiKonuk
Training and Research Hospital (OE), Istanbul, Turkey.
Supported, in part, by grants from the Research
Foundation of Istanbul University. Project number
2865/2008.
The authors have not disclosed any potential con-
flicts of interest.
For information regarding this article, E-mail:
evrensenturk2@yahoo.com
Copyright © 2012 by the Society of Critical Care
Medicine and Lippincott Williams & Wilkins
DOI: 10.1097/CCM.0b013e31823779ca