Complement-dependent immune complex-induced bronchial inflammation and hyperreactivity

Nicholas W. Lukacs1, M. Michael Glovsky2, and Peter A. Ward2

1 Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan 48109-0602; and 2 Asthma and Allergy Center, Huntington Hospital, Pasadena, California 91109-7013


    ABSTRACT
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

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 C5-deficient 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.

histamine; tumor necrosis factor; lung


    INTRODUCTION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

MULTIPLE PATHWAYS AND MECHANISMS 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 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
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

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 µg/mouse in 50 µl) 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.

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 (Delta Ptp) by the change in flow (Delta F; box pressure; Delta Ptp/Delta F in cmH2O · ml-1 · 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 µl 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-alpha 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 µl/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 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 µl/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
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

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 complement-dependent 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 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.


View larger version (25K):
[in this window]
[in a new window]
 
Fig. 1.   Immune complex-induced pulmonary inflammation mediated airway hyperreactivity. Mice were given intratracheal injections of anti-BSA (50 µg/mouse) and challenged with intravenous BSA (1 mg). The mice were subsequently examined for airway hyperreactivity responses after an intravenous injection of methacholine (100 µg/kg). Data are means ± SE from 5-6 mice/group and represent the change in airway resistance over background levels.



View larger version (136K):
[in this window]
[in a new window]
 
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 complex-induced 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.



View larger version (24K):
[in this window]
[in a new window]
 
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 µg/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.

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 anti-C5a-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.


View larger version (29K):
[in this window]
[in a new window]
 
Fig. 4.   Blockade of C5a decreased induction of airway hyperreactivity. Antibodies specific for C5a (50 µg/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 µg/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.

Association of airway hyperreactivity with TNF-alpha 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.


View larger version (25K):
[in this window]
[in a new window]
 
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.



View larger version (21K):
[in this window]
[in a new window]
 
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 C5-deficient 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-alpha 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 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 Fcgamma , have been engaged.


View larger version (37K):
[in this window]
[in a new window]
 
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.



View larger version (31K):
[in this window]
[in a new window]
 
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.


    DISCUSSION
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES

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-alpha 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 products. 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 beta 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-alpha , 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 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 C5-intact 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-alpha 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. Fcgamma 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.


    FOOTNOTES

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{at}umich.edu).

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.

Received 15 July 1999; accepted in final form 16 October 2000.


    REFERENCES
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
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[ISI][Medline].

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[ISI][Medline].

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[ISI][Medline].

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[ISI][Medline].

5.   Bertrand, C, and Geppetti P. Tachykinin and kinin receptor antagonists: therapeutic perspectives in allergic airway disease. Trends Pharmacol Sci 17: 255-259, 1996[ISI][Medline].

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[ISI][Medline].

7.   Busse, WW. Leukotrienes and inflammation. Am J Respir Crit Care Med 157: S210-S213, 1998[ISI].

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[Abstract/Free Full Text].

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[Abstract/Free Full Text].

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[Abstract/Free Full Text].

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[Abstract/Free Full Text].

12.   Cochrane, CG. The role of immune complexes and complement in tissue injury. J Allergy 42: 113-129, 1968[ISI][Medline].

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[ISI][Medline].

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[Abstract/Free Full Text].

15.   Curran, AD. The role of nitric oxide in the development of asthma. Int Arch Allergy Immunol 111: 1-4, 1996[ISI][Medline].

16.   Czermak, BJ, Sarma V, Pierson CL, Warner RL, Huber-Lang M, Bless NM, Schmal H, Friedl HP, and Ward PA. Protective effects of C5a blockade in sepsis. Nat Med 5: 788-792, 1999[ISI][Medline].

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[ISI][Medline].

18.   Drazen, JM. Leukotrienes as mediators of airway obstruction. Am J Respir Crit Care Med 158: S193-S200, 1998[Abstract/Free Full Text].

19.   FitzGerald, JM, and Macklem P. Fatal asthma. Annu Rev Med 47: 161-168, 1996[ISI][Medline].

20.   Ford-Hutchinson, AW. Activation of leukotriene production in granulocytes: effects on cell activation. Immunol Ser 57: 87-106, 1992[Medline].

21.   Gerard, C, and Gerard NP. C5A anaphylatoxin and its seven transmembrane-segment receptor. Annu Rev Immunol 12: 775-808, 1994[ISI][Medline].

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[ISI][Medline].

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[ISI][Medline].

24.   Hill, LD, Sun L, Leuschen MP, and Zach TL. C3 synthesis by A549 alveolar epithelial cells is increased by interferon-gamma and dexamethasone. Immunology 79: 236-240, 1993[ISI][Medline].

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[Abstract].

26.   Inman, RD, and Day NK. Immunologic and clinical aspects of immune complex disease. Am J Med 70: 1097-1106, 1981[ISI][Medline].

27.   Johnson, KJ, and Ward PA. Role of oxygen metabolites in immune complex injury of lung. J Immunol 126: 2365-2369, 1981[Free Full Text].

28.   Jose, PJ, Forrest MJ, and Williams TJ. Human C5a des Arg increases vascular permeability. J Immunol 127: 2376-2380, 1981[Abstract/Free Full Text].

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[Medline].

30.   Kilgore, KS, Shen JP, Miller BF, Ward PA, and Warren JS. Enhancement by the complement membrane attack complex of tumor necrosis factor-alpha -induced endothelial cell expression of E-selectin and ICAM-1. J Immunol 155: 1434-1441, 1995[Abstract].

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[ISI][Medline].

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[ISI][Medline].

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[ISI][Medline].

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[ISI][Medline].

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[Abstract].

36.   Lucchesi, BR. Complement activation, neutrophils, and oxygen radicals in reperfusion injury. Stroke 24, Suppl12: I41-I47, 1993[ISI][Medline].

37.   Lukacs, NW, and Ward PA. Inflammatory mediators, cytokines, and adhesion molecules in pulmonary inflammation and injury. Adv Immunol 62: 257-304, 1996[ISI][Medline].

38.   Manian, P. Genetics of asthma: a review. Chest 112: 1397-1408, 1997[Free Full Text].

39.   McFadden, ER, Jr. Exercise-induced airway obstruction. Clin Chest Med 16: 671-682, 1995[ISI][Medline].

40.   Meza, C, and Gershwin ME. Why is asthma becoming more of a problem? Curr Opin Pulm Med 3: 6-9, 1997[Medline].

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[ISI][Medline].

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[Abstract/Free Full Text].

43.   Mulligan, MS, Varani J, Warren JS, Till GO, Smith CW, Anderson DC, Todd RF, III, and Ward PA. Roles of beta 2 integrins of rat neutrophils in complement- and oxygen radical-mediated acute inflammatory injury. J Immunol 148: 1847-1857, 1992[Abstract/Free Full Text].

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[ISI][Medline].

45.   Ramos, BF, Zhang Y, and Jakschik BA. Neutrophil elicitation in the reverse passive Arthus reaction. Complement-dependent and -independent mast cell involvement. J Immunol 152: 1380-1384, 1994[Abstract/Free Full Text].

46.   Regal, JF. Role of the complement system in pulmonary disorders. Immunopharmacology 38: 17-25, 1997[ISI][Medline].

47.   Rossi, GL, and Olivieri D. Does the mast cell still have a key role in asthma? Chest 112: 523-529, 1997[Abstract/Free Full Text].

48.   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[ISI][Medline].

49.   Scherzer, H, and Ward PA. Lung injury produced by immune complexes of varying composition. J Immunol 121: 947-952, 1978[Abstract].

50.   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[ISI][Medline].

51.   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[Medline].

52.   Steinhilber, D. 5-Lipoxygenase: a target for antiinflammatory drugs revisited. Curr Med Chem 6: 71-85, 1999[ISI][Medline].

53.   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[ISI][Medline].

54.   Stimler, NP, Bloor CM, and Hugli TE. C3a-induced contraction of guinea pig lung parenchyma: role of cyclooxygenase metabolites. Immunopharmacology 5: 251-257, 1983[ISI][Medline].

55.   Stimler, NP, Hugli TE, and Bloor CM. Pulmonary injury induced by C3a and C5a anaphylatoxins. Am J Pathol 100: 327-338, 1980[Abstract].

56.   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[ISI][Medline].

57.   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[ISI][Medline].

58.   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[ISI][Medline].

59.   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[Abstract].

60.   Vogt, W. Anaphylatoxins: possible roles in disease. Complement 3: 177-188, 1986[Medline].

61.   Ward, PA. Leukotaxis and leukotactic disorders. A review. Am J Pathol 77: 520-538, 1974[Medline].

62.   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[Abstract].

63.   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[ISI][Medline].

64.   Wasserman, SI. Mast cells and airway inflammation in asthma. Am J Respir Crit Care Med 150: S39-S41, 1994[ISI][Medline].

65.   Williams, TJ, Hellewell PG, and Jose PJ. Inflammatory mechanisms in the Arthus reaction. Agents Actions 19: 66-72, 1986[ISI][Medline].

66.   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[Abstract].

67.   Wollina, U. Immune complexes---pathogenetic factors of autoimmune systemic lupus erythematosus. Allerg Immunol (Leipz) 30: 3-13, 1984[Medline].


Am J Physiol Lung Cell Mol Physiol 280(3):L512-L518
1040-0605/01 $5.00 Copyright © 2001 the American Physiological Society