Download Complement-dependent immune complex-induced - AJP-Lung

Am J Physiol Lung Cell Mol Physiol
280: L512–L518, 2001.
Complement-dependent immune complex-induced
bronchial inflammation and hyperreactivity
NICHOLAS W. LUKACS,1 M. MICHAEL GLOVSKY,2 AND PETER A. WARD2
1
Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan 48109-0602;
and 2Asthma and Allergy Center, Huntington Hospital, Pasadena, California 91109-7013
Received 15 July 1999; accepted in final form 16 October 2000
histamine; tumor necrosis factor; lung
syndrome, significant recruitment and activation of
leukocytes, and damage of vascular endothelial cells
and alveolar epithelial cells (27, 49, 62). These types of
events are observed in many diseases including autoimmune diseases and specific types of immune-mediated diseases such as allergic aspergillosis. In these
diseases, deposition of immune complexes is often associated with a severe hemorrhagic alveolitis in association with intense infiltration of neutrophils into the
interstitial and intra-alveolar compartments within
1–4 h (26, 37, 50, 67). Subsequently, the inflammatory
response results in the loss of integrity of the alveolar
capillary wall. Neutrophils recruited to the site are
activated and release a number of mediators, further
damaging the inflamed tissue. The response is dependent on the activation of a number of complement
proteins (45, 46). Inhibition of complement components
or complement activation products attenuates tissue
damage. Although the response in animal models of
IgG immune complex-induced inflammation in the
lung has been extensively studied, the ability of immune complex-induced responses to alter lung function
has not been well analyzed. In the present study, we
demonstrate that IgG immune complex-induced pulmonary inflammation induces airway hyperreactivity
in murine lungs in a manner that is complement dependent.
MATERIALS AND METHODS
can mediate the
induction of airway hyperreactivity (7, 8, 18, 47, 52).
Classic investigations have described IgE-mediated
mast cell activation followed by an intense late-phase
inflammatory response that includes the presence of
lymphocytes, monocytes, and eosinophils. Although
the mediators for acute and chronic airway responses
have not been completely identified, it has been established that the severity of the airway response is a
function of the intensity of the inflammation (7, 8, 18,
47, 52). Several specific and nonspecific events are
likely involved and contribute to the induction of airway hyperreactivity.
Acute lung injury that is induced by IgG immune
complex deposition in the lung includes a vascular leak
MULTIPLE PATHWAYS AND MECHANISMS
Address for reprint requests and other correspondence: N. W.
Lukacs, Univ. of Michigan Medical School, Dept. of Pathology, 1301
Catherine, Ann Arbor, MI 48109-0602 (E-mail: nlukacs@umich.edu).
L512
Animals. Female CBA/J mice were purchased from Jackson Laboratory (Bar Harbor, ME) and maintained under
standard pathogen-free conditions. C5-deficient mice
(B10.D2 “old line”) and C5-sufficient mice (B10.D2 “new
line”) were also obtained from the Jackson Laboratory and
are congenic mouse strains.
Induction of pulmonary immune complex inflammation.
Polyclonal rabbit anti-BSA IgG antibody (50 ␮g/mouse in 50
␮l) was injected intratracheally into CBA/J mice followed by
an injection of BSA (1 mg) via the tail vein. The mice were
euthanized at various time points post-BSA injection (1, 4,
and 24 h). The parameters examined included bronchoalveolar lavage (BAL) fluid content such as leukocytes, tumor
necrosis factor (TNF), and histamine and quantitation of
airway physiological changes.
The costs of publication of this article were defrayed in part by the
payment of page charges. The article must therefore be hereby
marked ‘‘advertisement’’ in accordance with 18 U.S.C. Section 1734
solely to indicate this fact.
1040-0605/01 $5.00 Copyright © 2001 the American Physiological Society
http://www.ajplung.org
Downloaded from http://ajplung.physiology.org/ by 10.220.33.2 on June 18, 2017
Lukacs, Nicholas W., M. Michael Glovsky, and Peter A.
Ward. Complement-dependent immune complex-induced bronchial inflammation and hyperreactivity. Am J Physiol Lung
Cell Mol Physiol 280: L512–L518, 2001.—Bronchoconstriction
responses in the airway are caused by multiple insults and
are the hallmark symptom in asthma. In an acute lung injury
model in mice, IgG immune complex deposition elicited severe airway hyperreactivity that peaked by 1 h, was maintained at 4 h, and was resolved by 24 h. The depletion of
complement with cobra venom factor (CVF) markedly reduced the hyperreactive airway responses, suggesting that
complement played an important role in the response. Blockade of C5a with specific antisera also significantly reduced
airway hyperreactivity in this acute lung model. Complement depletion by CVF treatment significantly reduced tumor necrosis factor and histamine levels in bronchoalveolar
lavage fluids, correlating with reductions in airway hyperreactivity. To further examine the role of specific complement
requirement, we initiated the immune complex response in
C5-sufficient and C5-deficient congenic animals. The airway
hyperreactivity response was partially reduced in the C5deficient mice. Complement depletion with CVF attenuated
airway hyperreactivity in the C5-sufficient mice but had a
lesser effect on the airway hyperreactive response and histamine release in bronchoalveolar lavage fluids in C5-deficient
mice. These data indicate that acute lung injury in mice after
deposition of IgG immune complexes induced airway hyperreactivity that is C5 and C5a dependent.
COMPLEMENT-DEPENDENT AIRWAY HYPERREACTIVITY
wash buffer, and cell-free supernatants were added (neat and
diluted 1:10) followed by incubation for 1 h at 37°C. Plates
were washed four times, streptavidin-peroxidase conjugate
(Bio-Rad, Richmond, CA) was added, and the plates were
incubated for 30 min at 37°C. Plates were washed again, and
chromogen substrate (Bio-Rad) was added and incubated at
room temperature to the desired extinction. The reaction was
terminated with 50 ␮l/well of 3 M H2SO4 solution, and the
plates were read at 490 nm in an ELISA reader. Standards
were 0.5 log dilutions of recombinant protein from 1 pg/ml to
100 ng/ml.
Statistical analysis. Significance was determined by
ANOVA, with P values ⬍ 0.05.
RESULTS
IgG immune complex-induced, complement-dependent airway hyperreactivity. Intrapulmonary deposition of IgG immune complexes can result in injury,
including a vascular leak syndrome, significant recruitment and activation of neutrophils, and damage to
lung tissue. To determine whether this complementdependent inflammatory response had a pathophysiological consequence, we examined lung function parameters in mice with IgG immune complex-induced
injury in the lung. Mice were injected intratracheally
with anti-BSA IgG followed by the intravenous injection of BSA (1 mg). As shown in Fig. 1, the induction of
airway hyperreactivity to a methacholine challenge
was most severe by 1 h, was still apparent but diminishing by 4 h, and had resolved by 24 h postimmune
complex deposition. Control mice given intratracheal
antibody with saline challenge instead of BSA showed
no significant increase in airway resistance. These
data demonstrated an early response to IgG immune
complexes that was characterized by increased airway
resistance. Histological changes in the lungs indicated
interstitial and intra-alveolar accumulations of neutrophils and were associated with the airway hyperreactivity changes observed (Fig. 2). Because the immune
complex-induced inflammation has been shown to be a
complement-mediated pathway, we examined mice
that had been complement depleted. This was accomplished with an injection of CVF 18 h before induction
Fig. 1. Immune complex-induced pulmonary inflammation mediated
airway hyperreactivity. Mice were given intratracheal injections of
anti-BSA (50 ␮g/mouse) and challenged with intravenous BSA (1
mg). The mice were subsequently examined for airway hyperreactivity responses after an intravenous injection of methacholine (100
␮g/kg). Data are means ⫾ SE from 5–6 mice/group and represent the
change in airway resistance over background levels.
Downloaded from http://ajplung.physiology.org/ by 10.220.33.2 on June 18, 2017
To deplete animals of complement, cobra venom factor
(CVF; 200 U/kg) was injected intraperitoneally the night
before surgery. This dose has previously been shown to reduce C3 levels in plasma to ⬍3% of normal levels (17, 34).
Measurement of airway hyperreactivity. Airway hyperreactivity was measured with a mouse plethysmograph specifically designed for the low tidal volumes (Buxco, Troy, NY) as
previously described (8–11). Briefly, the mouse to be tested
was anesthetized with pentobarbital sodium and intubated
via cannulation of the trachea with an 18-gauge metal tube.
The mouse was subsequently ventilated with a Harvard pump
ventilator (tidal volume ⫽ 0.4 ml, frequency ⫽ 120 breaths/
min, positive end-expiratory pressure ⫽ 2.5–3.0 cmH2O),
and the tail vein was cannulated with a 27-gauge needle for
injection of the methacholine challenge. The plethysmograph
was sealed, and readings were monitored by computer. Because the box was a closed system, a change in lung volume
was represented by a change in box pressure (Pbox) that was
measured by a differential transducer. The system was calibrated with a syringe that delivered a known volume of 2 ml.
A second transducer was used to measure the pressure swings
at the opening of the tracheal tube (Paw) referenced to the
body box (i.e., pleural pressure) and to provide a measure of
transpulmonary pressure (Ptp ⫽ Paw ⫺ Pbox). The tracheal
transducer was calibrated at a constant pressure of 20
cmH2O. Resistance was calculated with Buxco software by
dividing the change in pressure (⌬Ptp) by the change in flow
(⌬F; box pressure; ⌬Ptp/⌬F in cmH2O 䡠 ml⫺1 䡠 s⫺1) at two time
points from the volume curve based on a percentage of the
inspiratory volume. Once the mouse was hooked up to the box,
it was ventilated for 5 min before readings were acquired. Once
baseline levels were stabilized and initial readings were
taken, a methacholine challenge was given via the cannulated tail vein. After the determination of a dose-response
curve (0.001–0.5 mg), an optimal dose of 0.1 mg of methacholine was chosen. This dose was used throughout the rest of
the experiments in this study. After the methacholine challenge, the response was monitored, and the peak airway resistance was recorded as a measure of airway hyperreactivity.
Collection of BAL fluid. Lungs from mice were perfused
with 1 ml of PBS via intratracheal injection with a 1-ml
syringe and a 26-gauge needle. After 30–40 s, the PBS was
collected by aspiration with the same syringe and needle.
Between 700 and 800 ␮l could routinely be re-collected from
the perfused lung. The cells were then collected by centrifugation, resuspended in fresh PBS, and cytospun onto a glass
slide. The cytospins were then differentially stained with
eosin and hematoxylin. The percentage of cells was then
determined by counting the number of eosinophils per 200
total cells. Histamine levels were measured in cell-free BAL
fluid by ELISA with commercially available kits (Amac,
Westbrook, MA). Leukotrienes (LTs) from the BAL fluid were
assessed with enzyme immunoassay kits (Cayman Chemical,
Ann Arbor, MI).
Cytokine analysis by ELISA. The levels of TNF-␣ in BAL
fluid were measured by specific ELISA with a modification of
a double-ligand method as previously described (6). Briefly,
lung tissue was homogenized on ice with a tissue tearor
(Biospec Products, Racine, WI) for 30 s in 1 ml of PBS
containing 0.05% Triton X-100. The resulting supernatant
was isolated after centrifugation (10,000 g). Flat-bottom,
96-well microtiter plates (Nunc, Roskilde, Denmark) were
coated with 50 ␮l/well of rabbit polyclonal antibodies specific
for the cytokine or chemokine in question for 16 h at 4°C and
then washed with PBS and 0.05% Tween 20. Nonspecific
binding sites were blocked with 2% BSA in PBS and incubated for 90 min at 37°C. Plates were rinsed four times with
L513
L514
COMPLEMENT-DEPENDENT AIRWAY HYPERREACTIVITY
of the immune complex-induced response. The histological results indicated reduced inflammation in the
complement-depleted animals after immune complex
deposition (Fig. 2C), whereas in complement-intact
mice, a marked neutrophil infiltration was detected in
the bronchial wall (Fig. 2, B and D). The data presented in Fig. 3 clearly demonstrate that when complement was inhibited by pretreatment with CVF, the
airway hyperreactivity response was nearly abolished.
These results are consistent with previous data (21, 43,
57) that indicated that the immune complex-induced
airway responses require complement-mediated activation events and suggest that airway hyperreactivity
requires at least C3.
To further define the complement products that may
be responsible for the responses during intrapulmonary immune complex deposition, we treated the challenged mice with specific anti-C5a antibody given intratracheally. The data in Fig. 4 indicate that the
blockade of C5a significantly attenuated airway hyperreactivity responses. We examined the neutrophil recruitment responses and found a decrease in the antiC5a-treated mice compared with the control mice at 4 h
postdeposition (12.6 ⫾ 2.0 ⫻ 104 and 18.8 ⫾ 1.8 ⫻
104/ml, respectively; n ⫽ 5/group). Thus the immune
complex-induced airway hyperreactivity is dependent
on the activity of the complement, specifically, C5a.
Fig. 3. Complement depletion blocked the immune complex induction of airway hyperreactivity. Animals were given CVF (200 U/kg)
and challenged with immune complex-induced inflammation the
next morning. The mice were subsequently examined for airway
hyperreactivity responses after an intravenous injection of methacholine (100 ␮g/kg) 4 h after immune complex challenge. Bkgd,
background. Data are means ⫾ SE from 5–6 mice/group and represent the change in airway resistance over background levels. * P ⬍
0.05 compared with BSA/anti-BSA positive control.
Downloaded from http://ajplung.physiology.org/ by 10.220.33.2 on June 18, 2017
Fig. 2. Histological examination of immune complex-induced inflammation
inhibited by cobra venom factor (CVF).
Animals were given CVF (200 U/kg)
and challenged with immune complexinduced inflammation the next morning. Normal animals (A) were used for
comparison, and complement-depleted
animals (C) demonstrated substantially less inflammation compared with
control immune complex-challenged
mice (B and D). Arrow, neutrophils;
arrowhead, edema or leak from the inflammatory response.
COMPLEMENT-DEPENDENT AIRWAY HYPERREACTIVITY
Association of airway hyperreactivity with TNF-␣
and histamine. Because the early time point (1 h)
showed peak airway hyperreactivity, we examined
common cytokine pathways. TNF was significantly reduced after CVF-mediated complement depletion 1 h
postchallenge (Fig. 5). Measurement of histamine levels in the BAL fluid demonstrated that the mediator
was released into the airways 1 and 4 h postchallenge
(Fig. 6), producing levels of 8–10 and 25–30 nM, respectively. When animals were complement depleted,
significantly less histamine was detected in the BAL
fluid; at 1 and 4 h, the level was ⬍1 nM. When LTE4
was examined, there was a significant increase in its
levels in BAL fluid 1 h after the immune complex
response compared with that in control animals (412 ⫾
112 and ⬍10 pg/ml, respectively). However, no significant decrease in LTE4 was observed after CVF treatment (422 ⫾ 116 pg/ml). Thus at least two important
mediators were significantly affected by depletion of
complement before immune complex-induced inflammation, whereas levels of LTE4 were unaffected.
Fig. 5. CVF blockade of complement inhibited tumor necrosis factor
(TNF) production during immune complex-induced airway hyperreactivity. A 1-ml bronchoalveolar (BAL) fluid sample was harvested
from each mouse 1 h postchallenge, and the level of TNF was assayed
by specific ELISA. Data represent results from 5 different mice/
group. * P ⬍ 0.05.
Fig. 6. Immune complex-mediated inflammation induced histamine
release that is dependent on complement. Animals were given CVF
(200 U/kg) and challenged with immune complex-induced inflammation the next morning. Histamine levels were measured in cell-free
BAL fluid collected from individual mice. Data represent means ⫾
SE from 5–6 mice/group. * P ⬍ 0.05 compared with control.
Airway hyperreactivity in C5-deficient mice. We examined immune complex-induced inflammation in C5deficient mice compared with congenic C5-sufficient
mice. As shown in Fig. 7, airway hyperreactivity in
C5-deficient mice was similar to that in the C5-sufficient mice. The data indicated that the responses in the
B10 strain of mice shown in Fig. 7 were lower compared with those observed in the CBA/J strain (Fig. 1),
making comparisons between strains somewhat problematic. In these same studies, complement depletion
was carried out with an overnight treatment with CVF.
Induced complement depletion treatment significantly
lowered airway hyperreactivity responses in C5-sufficient mice, whereas the C5-deficient mice pretreated
with CVF showed no significant decrease (Fig. 7).
These data suggested that mechanisms other than
CVF-sensitive products were operative in the C5-deficient mice for the induction of airway hyperreactivity.
To correlate these results with previous observations
in Association of airway hyperreactivity with TNF-␣
and histamine, we also examined histamine levels in
the BAL fluids of these mice (Fig. 8). The C5-deficient
mice demonstrated a lower level of histamine in the
Fig. 7. Immune complex-induced pulmonary inflammation induced
airway hyperreactivity in C5-deficient mice that is not blocked by
CVF treatment. C5-sufficient and -deficient animals were given
CVF, and 18 h later, an immune complex challenge followed by
examination of the airway hyperreactivity response to methacholine.
Data are means ⫾ SE from 7–8 mice/group. * P ⬍ 0.05 compared
with BSA.
Downloaded from http://ajplung.physiology.org/ by 10.220.33.2 on June 18, 2017
Fig. 4. Blockade of C5a decreased induction of airway hyperreactivity. Antibodies specific for C5a (50 ␮g/mouse) were injected intratracheally along with the anti-BSA during the immune complex challenge. The mice were subsequently examined for airway
hyperreactivity responses after an intravenous injection of methacholine (100 ␮g/kg). Data are means ⫾ SE from 5–6 mice/group and
represent the change in airway resistance over background levels.
* P ⬍ 0.05 compared with BSA/anti-BSA positive control.
L515
L516
COMPLEMENT-DEPENDENT AIRWAY HYPERREACTIVITY
BAL fluid compared with the C5-sufficient mice after
immune complex responses. CVF treatment of C5-sufficient animals virtually abrogated the increase in histamine content in the BAL fluid, whereas CVF treatment only partially attenuated histamine release in
the BAL fluid of C5-deficient mice. These results partially correlate with the airway hyperreactivity data.
Collectively, the data suggest that IgG immune complex-induced airway hyperreactivity is C3, C5, and C5a
dependent. Increases in BAL fluid levels of TNF and
histamine (but not of LTE4) are complement dependent. In C5-deficient mice, there appears to be some C3
dependency, but other mediator pathways, such as
Fc␥, have been engaged.
DISCUSSION
Fig. 8. Histamine levels in the airway of C5-deficient mice were not
completely blocked by complement depletion with CVF. Histamine
levels were measured from cell-free BAL fluid collected in individual
mice 4 h after immune complex challenge. Data are means ⫾ SE
from 7–8 mice/group.
Downloaded from http://ajplung.physiology.org/ by 10.220.33.2 on June 18, 2017
Bronchial asthma is a disorder associated with airway inflammation and characterized pathophysiologically by airway hyperreactivity that is induced by a
variety of stimuli (5, 15, 32, 46, 59, 64). Classically,
asthmatic responses have been related to IgE-mediated pathways that induce mast cell and basophil activation and degranulation that lead to mediator release. In these present studies, we have demonstrated
that IgG immune complex deposition in the lung can
induce severe airway hyperreactivity in the mouse, a
finding that has not been previously reported. The
extensive body of information related to the pathophysiological mechanisms involved in this model of lung
injury has allowed us to obtain further information on
events that are triggered by tissue deposition of IgG
immune complexes. These reactions are accompanied
by TNF-␣ production, histamine release, and neutrophil accumulation. The hyperreactivity response
peaked early (by 1 h) and had resolved by 24 h. Interestingly, the airway permeability observed in this
model had previously been shown to peak at 4 h (42, 43,
63), suggesting that the hyperreactivity may be a related but separate event. Previous studies (21, 43, 57)
have clearly demonstrated that this inflammatory response is dependent on complement activation prod-
ucts. Depletion of complement by CVF inhibited the
development of airway hyperreactivity.
The complement activation products C3a and C5a
(anaphylatoxins) are known to produce airway smooth
muscle constriction in guinea pigs and humans (48, 54,
55, 60). C3a and C5a receptors have been found on
eosinophils, basophils, mast cells, monocytes, neutrophils, activated lymphocytes, and numerous nonmyeloid cell populations (21, 22). Anaphylatoxins can
cause histamine and mediator release from basophils
and mast cells (23, 25, 28, 31, 53, 65, 66). In addition,
C5a is a chemotactic and activating factor for effector
cells such as neutrophils and eosinophils (29, 36, 45,
57, 61). Finally, C3a and C5a have been shown to
upregulate the expression of selectins and ␤2-integrin
molecules (2, 30, 35, 41, 42). Endothelial cells respond
directly to C5a with increased expression of P-selectin
on their surfaces. In the present studies, it appears
that complement activation products lead to stimulation of mast cell/basophil populations, resulting in the
release of histamine, LTE4, and other acute mediators.
TNF-␣, which can be readily released from mast cells,
was also affected by depletion of the complement. Because histamine levels were more elevated at 4 h compared with 1 h, it is possible that multiple factors and
cell populations are involved in the progression of the
airway hyperreactivity responses. Taken together,
these studies suggest a significant role for complement
activation in the lung that leads to alteration of lung
function.
CVF (cobra C3b) is known to bind to factor B of the
alternative complement pathway, forming a stable
C3bBb complex and a markedly depleted C3, resulting
in blockade of all three complement pathways (3, 12,
13, 44). Pretreatment of mice with CVF to inhibit
complement-mediated pathways attenuates the immune complex-induced response within the lung. After
CVF injection, the hyperreactivity response to the
bronchospastic mediator methacholine was nearly
completely attenuated. This was further supported by
the histopathology in mice treated with CVF, which
demonstrated decreased injury, inflammation, and reduced airway changes. In addition, the depletion of
C5a with specific antisera further suggests that this
complement component is one of the key factors involved in the development of airway hyperreactivity
with this model. The data in this report demonstrate
that complement may be an essential pathway in the
hyperreactive airway response.
In an attempt to examine the requirements for specific complement components, we used congenic mice
that were either deficient or sufficient in C5. Although
there were some differences between the two groups in
their hyperreactive airway responses, the differences
were not significant. However, when these animals
were complement depleted with CVF, a distinct difference was observed. As noted above, treatment with
CVF attenuated the hyperreactivity response and reduced histamine levels in C5-sufficient but, surprisingly, not in C5-deficient mice. These results suggest
that although C3 and C5 play an important role in the
COMPLEMENT-DEPENDENT AIRWAY HYPERREACTIVITY
REFERENCES
1. Alpert SE, Pennington JE, and Colten HR. Synthesis of
complement by guinea pig bronchoalveolar macrophages. Effect
of acute and chronic infection with Pseudomonas aeruginosa. Am
Rev Respir Dis 129: 66–71, 1984.
2. Argenbright LW and Barton RW. Interactions of leukocyte
integrins with intercellular adhesion molecule-1 in the production of inflammatory vascular injury in vivo. The Shwartzman
reaction revisited. J Clin Invest 89: 259–272, 1992.
3. Ballow M and Cochrane CG. Two anticomplementary factors
in cobra venom: hemolysis of guinea pig erythrocytes by one of
them. J Immunol 103: 944–952, 1969.
4. Behera AK, Kumar M, Matsuse H, Lockey RF, and Mohapatra SS. Respiratory syncytial virus induces the expression of
5-lipoxygenase and endothelin-1 in bronchial epithelial cells.
Biochem Biophys Res Commun 251: 704–709, 1998.
5. Bertrand C and Geppetti P. Tachykinin and kinin receptor
antagonists: therapeutic perspectives in allergic airway disease.
Trends Pharmacol Sci 17: 255–259, 1996.
6. Burdick MD, Kunkel SL, Lincoln PM, Wilke CA, and Strieter RM. Specific ELISAs for the detection of human macrophage inflammatory protein-1 alpha and beta. Immunol Invest
22: 441–449, 1993.
7. Busse WW. Leukotrienes and inflammation. Am J Respir Crit
Care Med 157: S210–S213, 1998.
8. Campbell EM, Charo IF, Kunkel SL, Strieter RM, Boring
L, Gosling J, and Lukacs NW. Monocyte chemoattractant
protein-1 mediates cockroach allergen-induced bronchial hyperreactivity in normal but not CCR2-/- mice: the role of mast cells.
J Immunol 163: 2160–2167, 1999.
9. Campbell E, Hogaboam C, Lincoln P, and Lukacs NW.
Stem cell factor-induced airway hyperreactivity in allergic and
normal mice. Am J Pathol 154: 1259–1265, 1999.
10. Campbell E, Kunkel SL, Strieter RM, and Lukacs NW.
Temporal role of chemokines in a murine model of cockroach
allergen-induced airway hyperreactivity and eosinophilia. J Immunol 161: 7047–7053, 1998.
11. Campbell E, Kunkel SL, Strieter RM, and Lukacs NW.
Differential roles of IL-18 in allergic airway disease: induction of
eotaxin by resident cell populations exacerbates eosinophil accumulation. J Immunol 164: 1096–1102, 2000.
12. Cochrane CG. The role of immune complexes and complement
in tissue injury. J Allergy 42: 113–129, 1968.
13. Cochrane CG, Muller-Eberhard HJ, and Aikin BS. Depletion of plasma complement in vivo by a protein of cobra venom:
its effect on various immunologic reactions. J Immunol 105:
55–69, 1970.
14. Cole FS, Matthews WJ Jr, Marino JT, Gash DJ, and Colten
HR. Control of complement synthesis and secretion in bronchoalveolar and peritoneal macrophages. J Immunol 125: 1120–
1124, 1980.
15. Curran AD. The role of nitric oxide in the development of
asthma. Int Arch Allergy Immunol 111: 1–4, 1996.
16. Czermak BJ, Sarma V, Pierson CL, Warner RL, HuberLang M, Bless NM, Schmal H, Friedl HP, and Ward PA.
Protective effects of C5a blockade in sepsis. Nat Med 5: 788–792,
1999.
17. Dehring DJ, Steinberg SM, Wismar BL, Lowery BD, Carey
LC, and Cloutier CT. Complement depletion in a porcine
model of septic acute respiratory disease. J Trauma 27: 615–
625, 1987.
18. Drazen JM. Leukotrienes as mediators of airway obstruction.
Am J Respir Crit Care Med 158: S193–S200, 1998.
19. FitzGerald JM and Macklem P. Fatal asthma. Annu Rev Med
47: 161–168, 1996.
20. Ford-Hutchinson AW. Activation of leukotriene production in
granulocytes: effects on cell activation. Immunol Ser 57: 87–106,
1992.
21. Gerard C and Gerard NP. C5A anaphylatoxin and its seven
transmembrane-segment receptor. Annu Rev Immunol 12: 775–
808, 1994.
22. Glovsky MM, Hugli TE, Ishizaka T, Lichtenstein LM, and
Erickson BW. Anaphylatoxin-induced histamine release with
human leukocytes: studies of C3a leukocyte binding and histamine release. J Clin Invest 64: 804–811, 1979.
23. Hartman CT Jr and Glovsky MM. Complement activation
requirements for histamine release from human leukocytes: influence of purified C3ahu and C5ahu on histamine release. Int
Arch Allergy Appl Immunol 66: 274–281, 1981.
24. Hill LD, Sun L, Leuschen MP, and Zach TL. C3 synthesis by
A549 alveolar epithelial cells is increased by interferon-␥ and
dexamethasone. Immunology 79: 236–240, 1993.
25. Hook WA, Siraganian RP, and Wahl SM. Complement-induced histamine release from human basophils. I. Generation of
activity in human serum. J Immunol 114: 1185–1190, 1975.
Downloaded from http://ajplung.physiology.org/ by 10.220.33.2 on June 18, 2017
responses to immune complexes in complement-intact
mice, other pathways appear to compensate for the
absence of C5. It remains to be determined how this
compensation occurs, but it may involve several other
mediator pathways (1, 33, 58). Anti-C5a treatment
significantly blocked the airway hyperreactivity in C5intact mice, suggesting that this component plays a
significant role in the development of the responses.
This is in keeping with recently published results of a
study (16) that used anti-C5a antibody for the treatment of acute septic responses. The connection between mast cell/basophil activation and airway hyperreactivity is well accepted. Taken together, these
studies connect the interacting effects of complement
activation, mast cell/basophil degranulation, and
early-phase airway hyperreactivity, at least in complement-intact mice.
The bronchoconstrictive responses observed in
asthma are multifactorial and are triggered by numerous factors including allergens, cold air, exercise, pollutants, and viral infections (19, 38–40). Thus it is not
surprising that multiple mediators are involved in
pathophysiological responses during the evolution of
bronchospastic disease. The results from these studies
indicate that bronchospastic mediators that may be
common to allergen-induced responses are also produced during an immune complex-induced response.
Histamine and leukotrienes (LTC4, LTE4) as well as
TNF-␣ are produced at significant levels within the
airway during the immune complex response. Although the exact source of these mediators was not
identified in these studies, all of these can be produced
by mast cell/basophil populations within the lung.
Other cell populations, such as epithelial cells and
recruited inflammatory cell populations, have the ability to produce leukotrienes (4, 20, 51). Interestingly,
the C5-deficient mouse studies indicated that immune
complex-induced airway hyperreactivity is not completely dependent on this pathway. Fc␥ receptors are
also relevant and may be important in the airway
hyperreactivity response. Several studies (14, 24, 56)
have indicated that local lung populations may be a
source of complement proteins relevant in this injury.
In a global sense, this model demonstrates that both
complement-dependent and complement-independent
pathways may be relevant in acute bronchial hyperreactivity responses induced by IgG immune complexes.
Whether these responses translate directly to human
asthma needs to be considered and is worthy of further
investigation.
L517
L518
COMPLEMENT-DEPENDENT AIRWAY HYPERREACTIVITY
46.
47.
48.
49.
50.
51.
52.
53.
54.
55.
56.
57.
58.
59.
60.
61.
62.
63.
64.
65.
66.
67.
and -independent mast cell involvement. J Immunol 152: 1380–
1384, 1994.
Regal JF. Role of the complement system in pulmonary disorders. Immunopharmacology 38: 17–25, 1997.
Rossi GL and Olivieri D. Does the mast cell still have a key
role in asthma? Chest 112: 523–529, 1997.
Schellenberg RR and Foster A. In vitro responses of human
asthmatic airway and pulmonary vascular smooth muscle. Int
Arch Allergy Appl Immunol 75: 237–241, 1984.
Scherzer H and Ward PA. Lung injury produced by immune
complexes of varying composition. J Immunol 121: 947–952,
1978.
Segal AM, Calabrese LH, Ahmad M, Tubbs RR, and White
CS. The pulmonary manifestations of systemic lupus erythematosus. Semin Arthritis Rheum 14: 202–224, 1985.
Sjolander A, Schippert A, and Hammarstrom S. A human
epithelial cell line, intestine 407, can produce 5-hydroxyeicosatetraenoic acid and leukotriene B4. Prostaglandins 45: 85–96,
1993.
Steinhilber D. 5-Lipoxygenase: a target for antiinflammatory
drugs revisited. Curr Med Chem 6: 71–85, 1999.
Stephan V, Kuhr J, and Urbanek R. Anti-IgE- and complement-induced histamine release from peripheral leukocytes in
normals and atopics. Int Arch Allergy Appl Immunol 90: 326–
330, 1989.
Stimler NP, Bloor CM, and Hugli TE. C3a-induced contraction of guinea pig lung parenchyma: role of cyclooxygenase
metabolites. Immunopharmacology 5: 251–257, 1983.
Stimler NP, Hugli TE, and Bloor CM. Pulmonary injury
induced by C3a and C5a anaphylatoxins. Am J Pathol 100:
327–338, 1980.
Strunk RC, Eidlen DM, and Mason RJ. Pulmonary alveolar
type II epithelial cells synthesize and secrete proteins of the
classical and alternative complement pathways. J Clin Invest 81:
1419–1426, 1988.
Till GO, Johnson KJ, Kunkel R, and Ward PA. Intravascular activation of complement and acute lung injury; dependency
on neutrophils and toxic oxygen metabolites. J Clin Invest 69:
1126–1135, 1982.
Tulunay M, Demiralp S, Tastan S, Akalin H, Ozyurda U,
Corapcioglu T, and Akarsu ES. Complement (C3, C4) and
C-reactive protein responses to cardiopulmonary bypass and
protamine administration. Anaesth Intensive Care 21: 50–55,
1993.
Umetsu DT and DeKruyff RH. Th1 and Th2 CD4⫹ cells in the
pathogenesis of allergic diseases. Proc Soc Exp Biol Med 215:
11–20, 1997.
Vogt W. Anaphylatoxins: possible roles in disease. Complement
3: 177–188, 1986.
Ward PA. Leukotaxis and leukotactic disorders. A review. Am J
Pathol 77: 520–538, 1974.
Warren JS, Kunkel SL, Johnson KJ, and Ward PA. In
vitro activation of rat neutrophils and alveolar macrophages
with IgA and IgG immune complexes. Implications for immune complex-induced lung injury. Am J Pathol 129: 578–
588, 1987.
Warren JS, Yabroff KR, Remick DG, Kunkel SL, Chensue
SW, Kunkel RG, Johnson KJ, and Ward PA. Tumor necrosis
factor participates in the pathogenesis of acute immune complex
alveolitis in the rat. J Clin Invest 84: 1873–1882, 1989.
Wasserman SI. Mast cells and airway inflammation in asthma.
Am J Respir Crit Care Med 150: S39–S41, 1994.
Williams TJ, Hellewell PG, and Jose PJ. Inflammatory mechanisms in the Arthus reaction. Agents Actions 19: 66–72, 1986.
Williams TJ and Jose PJ. Mediation of increased vascular
permeability after complement activation. Histamine-independent action of rabbit C5a. J Exp Med 153: 136–153, 1981.
Wollina U. Immune complexes—pathogenetic factors of autoimmune systemic lupus erythematosus. Allerg Immunol (Leipz) 30:
3–13, 1984.
Downloaded from http://ajplung.physiology.org/ by 10.220.33.2 on June 18, 2017
26. Inman RD and Day NK. Immunologic and clinical aspects of
immune complex disease. Am J Med 70: 1097–1106, 1981.
27. Johnson KJ and Ward PA. Role of oxygen metabolites in
immune complex injury of lung. J Immunol 126: 2365–2369,
1981.
28. Jose PJ, Forrest MJ, and Williams TJ. Human C5a des Arg
increases vascular permeability. J Immunol 127: 2376–2380,
1981.
29. Kay AB. Studies on eosinophil leucocyte migration. II. Factors
specifically chemotactic for eosinophils and neutrophils generated from guinea-pig serum by antigen-antibody complexes. Clin
Exp Immunol 7: 723–737, 1970.
30. Kilgore KS, Shen JP, Miller BF, Ward PA, and Warren JS.
Enhancement by the complement membrane attack complex of
tumor necrosis factor-␣-induced endothelial cell expression of
E-selectin and ICAM-1. J Immunol 155: 1434–1441, 1995.
31. Kings M, Nydegger UE, and de Weck AL. Control of immune
complex and zymosan-mediated anaphylatoxin generation by
proteins B and H of the alternative complement pathway. Immunology 51: 123–131, 1984.
32. Kraneveld AD, Folkerts G, Van Oosterhout AJ, and
Nijkamp FP. Airway hyperresponsiveness: first eosinophils
and then neuropeptides. Int J Immunopharmacol 19: 517–
527, 1997.
33. Langlois PF and Gawryl MS. Accentuated formation of the
terminal C5b-9 complement complex in patient plasma precedes
development of the adult respiratory distress syndrome. Am Rev
Respir Dis 138: 368–375, 1988.
34. Larsen GL, Presley DM, Graves JP, and Giclas PC. The
effect of intravascular complement activation and brief episodes
of hypoxia on protein in bronchoalveolar lavage fluid in C5
sufficient and deficient mice. Pediatr Pulmonol 11: 302–309,
1991.
35. Lozada C, Levin RI, Huie M, Hirschhorn R, Naime D,
Whitlow M, Recht PA, Golden B, and Cronstein BN. Identification of C1q as the heat-labile serum cofactor required for
immune complexes to stimulate endothelial expression of the
adhesion molecules E-selectin and intercellular and vascular cell
adhesion molecules 1. Proc Natl Acad Sci USA 92: 8378–8382,
1995.
36. Lucchesi BR. Complement activation, neutrophils, and oxygen
radicals in reperfusion injury. Stroke 24, Suppl 12: I41–I47,
1993.
37. Lukacs NW and Ward PA. Inflammatory mediators, cytokines,
and adhesion molecules in pulmonary inflammation and injury.
Adv Immunol 62: 257–304, 1996.
38. Manian P. Genetics of asthma: a review. Chest 112: 1397–1408,
1997.
39. McFadden ER Jr. Exercise-induced airway obstruction. Clin
Chest Med 16: 671–682, 1995.
40. Meza C and Gershwin ME. Why is asthma becoming more of
a problem? Curr Opin Pulm Med 3: 6–9, 1997.
41. Mulligan MS, Johnson KJ, Todd RF III, Issekutz TB, Miyasaka M, Tamatani T, Smith CW, Anderson DC, and
Ward PA. Requirements for leukocyte adhesion molecules in
nephrotoxic nephritis. J Clin Invest 91: 577–587, 1993.
42. Mulligan MS, Smith CW, Anderson DC, Todd RF III, Miyasaka M, Tamatani T, Issekutz TB, and Ward PA. Role of
leukocyte adhesion molecules in complement-induced lung injury. J Immunol 150: 2401–2406, 1993.
43. Mulligan MS, Varani J, Warren JS, Till GO, Smith CW,
Anderson DC, Todd RF III, and Ward PA. Roles of ␤2
integrins of rat neutrophils in complement- and oxygen radicalmediated acute inflammatory injury. J Immunol 148: 1847–
1857, 1992.
44. Newman S, Glovsky MM, Ghekiere L, and Alenty A. Quantitative requirements for C3 to induce Forssman systemic shock
and cutaneous hemorrhagic vasculitis in guinea pigs. J Allergy
Clin Immunol 59: 327–333, 1977.
45. Ramos BF, Zhang Y, and Jakschik BA. Neutrophil elicitation
in the reverse passive Arthus reaction. Complement-dependent