Inactivation of Chemotactic Factor Inactivator by Cigarette Smoke A Potential Mechanism of Modulating Neutrophil Recruitment to the Lung 1 - 3

RICHARD A. ROBBINS, GAIL L. GOSSMAN, KENNETH J. NELSON, SEKIYA KOYAMA, AUSTIN B. THOMPSON, and STEPHEN I. RENNARD

Introduction

Cigarette smoking is associated with an influx of neutrophils into the lung (1-8). Current concepts suggest that these neutrophils contribute to the lung damage secondary to cigarette smoking by releasing proteases or oxygen radicals that damage lung tissue (9-11). Thus, the mechanism(s) accounting for the recruitment of neutrophils into the lung is likely important in understanding the pathogenesis of smoke-induced lung disease. One potential mechanism for recruitment of neutrophils is activation of the complement system. In support of the importance of complement activation in the neutrophil recruitment to the lung, smoke-exposed mice deficient in the fifth component of complement (C5) recruit fewer neutrophils to the lung than do C5sufficient mice (12). In addition, cigarette smoke can activate the complement system (13-15). Activated complement components can also activate alveolar macrophages to release additional chemotactic factors for neutrophils, thus amplifying neutrophil recruitment (16-18). Complement activation results in chemotactic activity by cleavage of C5, releasing the potent neutrophil chemotactic factor C5a (19-21). C5a can be converted into the less potent chemotactic factor C5a des Arg by the action of carboxypeptidase N (22). Recent evidence suggests that the chemotactic activity of C5a or C5a des Arg can be modulated by GcGlobulin (GcG), a vitamin-Dbinding protein (23, 24). GcG can function as a cochemotaxin for C5a or C5a des Arg by binding to the chemotactic factors and enhancing their chemotactic potency. Chemotactic factor inactivator (CFI) can function as an inhibitor of C5a-directed neutrophil chemotaxis by binding to GcG and preventing GcG from binding to C5a or C5a des Arg, thus

SUMMARY Activation of the complement system with generation of the potent neutrophil chemotactic factor C5a has been proposed to play a significant role in the neutrophil accumulation in the lungs of cigarette smokers. Chemotactic factor inactivator (CFI) can inhibit C5a-directed neutrophil chemotaxis by binding to the C5a cochemotaxin GcGlobulin (GcG), a vitamin-D-binding protein, and inhibiting the capacity of GcG to enhance the chemotactic activity of C5a. Because cigarette smoke can inhibit the function of some proteins, a loss of CFI functional activity induced by cigarette smoke would allow an increased caPacity of GcGto augment C5a-directed neutrophil chemotaxis. In order to test this hypothesis, cigarette smoke was bubbled through a CFI solution, and the solution was evaluated for its ability to inhibit the chemotactic activity of C5a and GcG. Smoke-treated CFI inhibited only 36% of the C5a-GcG chemotactic activity. In contrast, a CFI solution treated with air inhibited 62% of the chemotactic activity (p < 0.001).Consistent with these observations, smoketreated CFI exhibited a decreased capacity to bind to GcG and a decreased capacity to inhibit the binding of C5a des Arg to GcG. CFI contained in the bronchial lavage fluids obtained from patients with chronic obstructive pulmonary disease secondary to cigarette smoking and asymptomatic smokers exhibited a decreased capacity to inhibit C5a-GcG neutrophil chemotaxis and to bind to GcG (p < 0.05, both comparisons). Furthermore, smoke bubbled through normal bronchial lavage fluid decreased the capacity of CFI to bind to GcG. These data suggest that one mechanism accounting for an influx of neutrophils into the lungs of cigarette smokers may be a loss of functional activity of CFI secondary to cigarette smoke. AM REV RESPIR DIS 1990; 142:763-768

decreasing the potency of the chemotactic factors (25). CFI is a heat- and pH-labile protein with an approximate molecular weight of 90 kD (26, 27). CFI has also been previously identified in the lower respiratory tract (28). Because cigarette smoke can functionally inactivate other proteins in the lower respiratory tract (29), we hypothesized that cigarette smoke could inactivate CFI. Presumably, this would lead to an increased accumulation of neutrophils within the lung by allowing enhanced neutrophil chemotaxis secondary to complement activation. In this report, evidence is presented that cigarette smoke can inactivate CFI and that CFI is inactivated in the lower respiratory tract of cigarette smokers. Methods CFI Preparation For use in the binding experiments with GcG, highly purified CFI was prepared from normal human serum, as previously described,

by sequential ammonium sulfate precipitation, gel filtration (Sephacryl S-200; Pharmacia, Piscataway, NJ), chromatofocusing (PBE 94; Pharmacia), and hydrophobic column chromatography (eo-hexyl agarose; Miles, Inc., Naperville, IL) (28). The CFI preparation yielded a single band on a reduced 7.5070 polyacrylamide gel stained with Coomassie brilliant blue. The amount of CFI in the preparation was quantified using an enzyme-linked immunosorbent assay (ELISA) as previously described (28). (Received in originalform September 22, 1989 and in revised form March 8, 1990) 1 From the Research Service, Omaha Veterans Administration Medical Center, and the Pulmonary and Critical Care Section, Department of Internal Medicine, University of Nebraska Medical Center, Omaha, Nebraska. 2 Supported by grants from the Nebraska State Department of Health and the VeteransAdministration. 3 Correspondence and requests for reprints should be addressed to Richard A. Robbins, M.D., Pulmonary and Critical Care Section, Department of Internal Medicine, University of Nebraska Medical Center, 42nd and DeweyAvenue, Omaha, NE 68105.

763

764

For use in the chemotaxis assays, CFI was partially purified from normal human serum or from bronchial lavage fluid by fractional precipitation at 45 and 65070 ammonium sulfate saturation (26, 27). The pellet was dissolved in distilled water at the original volume for serum or at one-tenth the original volume for the lavage fluid. The serum or lavage fluid was dialyzed overnight against phosphate-buffered saline (PBS). This ammonium sulfate fraction (45 to 65070) contains CFI as the predominant inhibitor of GcG cochemotaxin activity for C5a (28).

GcG Preparation GcG waspurchased from a commercial source (Sigma, St. Louis, MO). The preparation yielded a single band on 12.5070 polyacrylamide gelsstained with silver(Sigma)and contained all three isoforms of GcG by isoelectric focusing (Phast System; Pharmacia). C5a Preparation For use in the chemotaxis assays, partially purified C5a was prepared according to the methods of Kreutzer and coworkers (30). Briefly, C5a was generated in whole human serum by incubation with zymosan (Sigma) at 37° C in the presence of 1.0 M epsilonaminocaproic acid (EACA) (Sigma) and fractionated over Sephadex G-75 (Pharmacia). The C5a-containing fractions wereidentified, and the amount of C5a was quantified by radioimmunoassay (Amersham, Inc., Arlington Heights, IL). Partially purified C5a prepared in this manner would be expected to contain GcG (24, 25), and the preparation contained GcG at a concentration of 17.5 ug/ml quantified by ELISA. The fractions were frozen until used and were dialyzed against PBS for at least 2 hours prior to usage in the chemotaxis assays. Radioactive C5a des Arg radiolabeled with 125 1(specificactivity, approximately 4.2 x 105 cpm/J.1g C5a) was purchased (Amersham) for use in the binding assays. The radiolabeled C5a des Arg yielded a single band on autoradiography of a 12.5070 polyacrylamide gel. Preparation of Smoke-exposed CFI A one-milliliter solution of either purified or crude CFI was exposed to cigarette smoke by bubbling smoke drawn from a lit cigarette (Camel unfiltered; R.J. Reynolds Co., Winston-Salem, NC) in an apparatus similar to that described by Carp and Janoff (29). One 30-ml puff of cigarette smoke was drawn over 20 s everyminute and bubbled slowlythrough the solution over 20 to 30 s using a 40-cm tube to avoid heating. Sevenpuffs weredrawn from each cigarette. As a control, air was bubbled through a CFI solution under identical conditions, except the cigarette was unlit. The CFI solution was dialyzed for 2 to 4 h prior to usage because undialyzed smoke extract markedly inhibited neutrophil chemotaxis. For some experiments, a smoke extract was prepared as above by substituting PBS in place of the CFI preparation.

ROBBINS, GOSSMAN, NELSON, KOYAMA, THOMPSON, AND RENNARD

Inhibition of C5a-GcG Neutrophil Chemotaxis by CFI Functional activity of CFI was determined as previously described (26-28). Briefly, the smoke-exposed CFI or air-exposed CFI was incubated with an equal volume of the C5a and GcG solution for 30 min at 37° C. To quantify the chemotactic activity of the samples, a blindwell chamber method was used. The chemotactic stimuli were placed in the lower wells of a 48-wellchemotaxis assembly and covered with a polycarbonate membrane with 3 urn pores. Fifty microliters of purified human neutrophils at a concentration of 3 x 106 cells/ml in Gey's medium with 2070 bovine serum albumin (Sigma) and 225 U/ml of penicillin and 225 ug/ml of streptomycin were added to the upper wells. The neutrophils were allowed to migrate for 30 min at 37° C in a 5070 CO 2 humidified atmosphere. The membranes werethen removed, fixed and stained (Diff-Quiks; American Hospital Supply, McGraw Park, IL), and mounted on glass slides. Chemotactic activity was quantified by counting the number of cells that completely migrated through the membrane. Five high power fields (HPF x 1,000) were counted from each duplicate well. Gey's medium alone served as a negative control, and normal human serum complement activated with lipopolysaccharide (Escherichia coli 0127:B8; Difco, Inc., Detroit, MI) and diluted 1:10 with PBS served as a positive control. The percent inhibition was calculated by the method of Berenberg and Ward (26) as 100 x 1 - [ChemotaxiS of sample - Chemotaxis of GeY's]. Chemotaxis of C5a - Chemotaxis of Gey's

Interaction of CFI and GcG Because previous investigations have determined that CFI binds to GcG (25), the capacity of cigarette smoke to alter CFI and GcG interaction was investigated using a "sandwich" ELISA. GcG was dissolved in Voller's buffer (0.02 carbonate buffer at pH 9.6) at 4 ug/ml, Two-tenths of a milliliter was placed in each well of a flat-bottomed polystyrene microtiter plate (Nunc-Immuno II; USA/Scientific Plastics, Ocala, FL), and the GcG was allowedto adsorb to the plastic overnight at 4°C. After incubation, the GcGcoated plates were washed three times with PBS-Tween® (0.02 sodium phosphate, 0.15M sodium chloride, 0.05070 Tween-20). Two tenths of a milliliter of smoke-exposed or airexposed CFI was added to the GcG-coated wells in serial 1:3dilutions and allowed to incubate at 22° C for 30 min to allow the CFI to bind to the GcG. The plates were again washed three times with PBS-Tween,and 200 ul of monospecific rabbit antihuman CFI antiserum diluted 1:500 with PBS-Tween were added to each well to detect the bound CFI. After 30 min of incubation at 22° C, the plates were washed again with PBS-Tween, and the IgG bound to the CFI was detected by the addition of 200 J.11 of a 1:500 dilution of

peroxidase-conjugated goat antirabbit IgG (Sigma) to each well.After 90 min of incubation, the plates wereagain washed three times, and peroxidase activity was detected by the addition of 200 ul of o-phenylenediamine (100 ug/ml in 0.01070 H 2 0 2 ) . The reaction was visually monitored, and after approximately 30 min, 25 ul of 8N sulfuric acid were added to each well to terminate the reaction. Absorbance of each well was measured at 490 nm, and the CFI -GcG was expressed as the optical density.

Inhibition of GcG and C5a Interaction by CFI Because GcG functions as a cochemotaxin for C5a or C5a des Arg by binding to the C5a (23,24) and CFI functions as an inhibitor of C5a-directed neutrophil chemotaxis by preventing GcG-C5a binding (25), a loss of CFI activity should result in an increased capacity of C5a to bind to GcG. To test this, GcG was dissolved in Voller's buffer at 4 ug/ml, and 200 ul were added to a 96-well plate with removable wells(Removawells; Dynatech Laboratories, Inc., Alexandria, VA) and allowed to adsorb to the plastic overnight at 4° C. After washing three times with PBS to remove unbound GcG, 200 ul of smoke-exposed or air-exposed CFI were added to the plate. After 30 min of incubation, 200 J.11 of 125 1 C5a des Arg (6,000 dpm) were added to each well and allowed to incubate 1 h at 22° C. The wells were washed with room temperature PBS, separated, and counted in a gamma counter. C5a des Arg binding was expressed as the mean dpm for each quadruplicate well. Bronchial Lavage The above experiments suggested that CFI could be functionally inactivated by cigarette smoke. To determine if these in vitro observations might have in vivo significance, the lower respiratory tract was sampled by bronchoscopy and bronchial lavage from four normal nonsmokers, two smokers with normal physical exam results and pulmonary function tests, and seven heavy smokers (> 2 packs/day) with chronic obstructive pulmonary disease who met ATScriteria for chronic bronchitis. Patients were permitted to smoke until the time of their bronchoscopy. After obtaining informed consent, local lidocaine anesthesia was obtained by nebulization and spraying of the upper airway, and the subjects were: sedated intravenously (meperidine 25 to 100mg and diazepam 1 to 10 mg). A fiberoptic bronchoscope (Model IT-R or PD-IO; Olympic Corp. of America, New Hyde Park, NY) was passed transorally and wedged in a subsegmental bronchus of the right middle lobe, right lower lobe, left lower lobe, or lingula. Twenty milliliters of normal saline were infused and gently aspirated from three separate sub segments. The recovered lavage fluid from an initial 20-ml aliquot of saline appears enriched for bronchial cells and proteins compared with later aliquots and, therefore, the recovered lavage

INACTIVATION OF CHEMOTACTIC FACTOR INACTIVATOR BY CIGARETTE SMOKE

has been termed bronchial lavage fluid (8). The bronchial lavages were then pooled, passed through a nylon mesh to remove mucus, and centrifuged (500 g for 5 min) to remove cells, and the supernatant bronchial lavage fluid was frozen at - 80° C until used.

CFI Functional Activity in Bronchial Lavage The CFI was expressed as ng/ml and also as ug/ml albumin. Albumin was measured using a previously described ELISA (28). Concen tration of CFI was measured in the bronchial lavage fluid by a previously described ELISA (28). CFI was partially purified from the bronchiallavage fluids as described above by ammonium sulfate precipitation. The lavage fluids were adjusted to equal CFI concentrations by dilution with PBS. The diluted bronchial lavages were then combined with an equal volume of C5a-OcO and the capacity of CFI to inhibit neutrophil chemotaxis was determined as described above. The capacity of the bronchial CFI to bind to OcO was also evaluated. This was performed by addition of the bronchial lavage fluids to OcO-coated ELISA plates and quantifying the CFI binding using the sandwich ELISA described above. Inactivation of Normal Bronchial Lavage CFI by Cigarette Smoke To determine if cigarette smoke could inactivate bronchial lavage CFI, bronchial lavages from three normal subjects were exposed to smoke from six cigarettesor to air as described

above. The CFI was then evaluated for its capacity to bind to OcO by the sandwich ELISA described above.

ed compared with that of air-exposed CFI, and it decreased in a dose-dependent manner (figure 2).

Statistical Analysis All data were expressedas mean ± SEM. Student's unpaired two-tailed t test was used for statistical comparisons. Significance was defined as p < 0.05.

Inhibition of GcG-C5a Interaction by CFI CFI readily inhibited the binding of 1251 C5a des Arg to GcG. Cigarette smoke from sixcigarettes significantly decreased the capacity of CFI to inhibit this interaction (p < 0.01) (figure 3).

Results

Inhibition of C5a-GcG Neutrophil CFI Functional Activity in Chemotaxis by CFI Bronchial Lavage CFI readily inhibited C5a-GcG neutrophil chemotaxis, and the capacity of CFI CFI was antigenically elevated in bronto inhibit this chemotactic activity was chiallavage obtained from smokers comreduced by cigarette smoke in a dose- pared with that obtained from nonsmokdependent manner (p < 0.01, three ciga- ers whether expressed as ng/ml (91 ± 23 rettes and six cigarettes versus air) (fig- versus 11 ± 2, p < 0.01) or as ug/mg alure 1). Undialyzed cigarette smoke ex- bumin (0.96 ± 0.30 versus 0.29 ± 0.07, tract markedly inhibited neutrophil p < 0.05). When partially purified CFI chemotaxis, but it had no effect when di- from the bronchial lavagefluids obtained alyzed for> 2 h (data not shown) . Dia- from normal subjects was adjusted to lyzed cigarette smoke extract had no ef- equal CFI concentrations, the CFI resultfect on the capacity of CFI to inhibit neu- ed in a modest inhibition of C5a-GcG trophil chemotaxis, suggesting that (figure 4). In contrast, there was no inhiinactivation might depend on the gas bition in the partially purified CFI obphase of cigarette smoke. tained from the smokers' bronchial lavage fluids adjusted to the same CFI conInhibition of CFI-GcG Interaction centrations (p = 0.001, compared with by Cigarette Smoke that from normal subjects) (figure 4). The binding of smoke-exposed CFI to To confirm the above results, the caGcG was decreased at each dilution test- pacity of bronchial lavage fluid CFI to

A.

B.

100

Z a >= en

0..

:::

-J -J

0 .4

80

~ (/)

765

80

I

~

E c: 0 .3

60

~

UJ

0

....

~ >-

OJ

z o

~

UJ

.... 60

ffi

>

I-

40

>

!:>

o ..:

>-

Z

s

0

....

Ci5

.... >= ~

UJ

o

-J

l-



Ci5

z w

0

-J

o o



0.2

o

-20

i=

0

0

c

0)

«

~

u,

E 0

Q.

0 Nonsmokers

o

Smokers

0.1

Fig. 4. Inactivation of chemotactic factor inactivator (CFI) in the lower respiratory tract of cigarette smokers. CFI functional activity expressed as the percent inhibition of C5a-GcGlobulin chemotaxis is on the vertical axis, and the SUbjects grouped according to smoking status are on the horizontal axis.

8 o Nonsmokers

lung. The presence of increased numbers of neutrophils in the lower respiratory tract has been demonstrated both in human smokers and in animals exposed to cigarette smoke (1-8). Intratracheal instillation of neutrophil elastase in experimental animals results in lung destruction similar to emphysema (9, 10). Furthermore, neutrophils can release oxidants, which can further augment lung destruction (31). Several mechanisms that may result in the influx of neutrophils into the lower respiratory tract have been proposed, including chemotactic factors in cigarette smoke (32),activation of cellsin the lower respiratory tract to release chemotactic factors (2), and activation of the complement system (13-15, 33). Activation of the complement system as a potential source of neutrophil chemotactic activity in smoking-associated lung disease is supported by several lines of evidence. First, mice deficient in C5 attract fewer neutrophils into the lung than do C5-sufficient mice when exposed to cigarette smoke (12). Second, cigarette smoke has been demonstrated to activate the complement system (13-15). Third, exposure of normal serum to cigarette smoke results in both the generation of C5a and increased neutrophil chemotactic activity (Robbins RA, unpublished observations). Complement activation results in the cleavage of C5 to release the potent chemotactic peptide C5a (19-21). In the

Smokers

Fig. 5. Chemotactic factor inactivator (CFI) and GcGlobulin interaction using CFI obtained from the lower respiratory tract by bronchial lavage. The capacity of CFI to interact with GcGlobulin is expressed on the vertical axis as optical density at 490 nm, and the subjects grouped according to smoking status are on the horizontal axis.

Ec:

0.12

0 0>

~

.->-

(j)

0.08

z

w

C

...J

-e

0

0.04 i= Q.. 0

1:

1~3

1:9

1:27

DILUTION BRONCHIAL LAVAGE FLUID Fig. 6. Capacity of cigarette smoke to inactivate chemotactic factor inactivator (CFI) from normal bronchial lavage fluid. CFI binding to GcGlobulin is expressed on the vertical axis as optical density at 490 nm and the dilutions of bronchial lavage obtained from normal nonsmokers is on the horizontal axis. The solid symbols represent bronchial lavage exposed to air, and the open symbols represent bronchial lavage exposed to six cigarettes. The symbols with the same shape indicate matching samples.

INACTIVATION OF CHEMOTACTIC. FACTOR INACTIVATOR BY CIGARETTE SMOKE

serum, C5a can be rapidly degraded to the less potent chemotactic factor C5a des Arg by the action of carboxypeptidase N (22). However,recent evidence has suggested that the chemotactic potency of C5a des Arg and C5a may be enhanced by the cochemotaxin GcG, which binds to C5a or C5a des Arg (23, 24). GcG has been identified in the lower respiratory tract (34), and recent evidence suggests that GcG obtained from the lower respiratory tract can function as a cochemotaxin (35). CFI can inhibit C5a-directed neutrophil chemotaxis by binding GcG and preventing its binding to C5a (25). It is known that cigarette smoke can alter the function of proteins in the lower respiratory tract. For example, alphal-antiprotease can be inactivated by cigarette smoke oxidation of a methionine residue at its active site (26, 36, 37). However, the functional activity of all proteins is not altered by cigarette smoke. In the context of the present study, preliminary evidence indicates that smokeexposed GcG can bind C5a des Arg as readily as GcG exposed only to air (35). Anadditional mechanism of neutrophil recruitment into the lung is the re"lease of neutrophil chemotactic factor(s) by alveolar macrophages. In support of this concept, Hunninghake and Crystal (2) have demonstrated that smoke can stimulate normal alveolar macrophages to release chemotactic activity and that alveolar macrophages obtained from cigarette smokers spontaneously release increased quantities of chemotactic factor(s). CFI has been demonstrated to attenuate alveolar macrophage release of chemotactic activity by a variety of stimuli (18). Although not evaluated in the present study, it is possible that inactivation of CFI by cigarette smoke might allow increased release of chemotactic factors from alveolar macrophages. The present study did not definitely demonstrate a net loss of CFI activity in the lower respiratory tract of cigarette smokers. The CFI obtained from the lower respiratory tract was adjusted to equal CFI concentrations. Because the concentration of many proteins appear increased in the lungs of cigarette smokers (34), it is possible that an increase in CFI either by increased local synthesis or increased diffusion from the vasculature might abrogate the functional inactivation of CFI. Lung disease secondary to cigarette smoking remains an important health problem. The present study demonstrates that cigarette smoke can inactivate CFI, which may contribute to an increased ac-

cumulation of neutrophils in the lower respiratory tract of cigarette smokers. Because these neutrophils likely contribute to the lung damage induced by cigarette smoking, CFI inactivation in the lower respiratory tract may contribute to the multitude of harmful effects of cigarette smoke that eventually result in lung destruction. Acknowledgment The writers acknowledge the expert secretarial assistance of Phyllis Siracusano in preparation of the manuscript. References 1. Niewoehner DE, Klernerman, Rice DB. Pathologic changes in the peripheral airways of young cigarette smokers. N Engl J Med 1974; 291:755-8. 2. Hunninghake GW, Crystal RG. Cigarette smoking and lung destruction. Accumulation of neutrophils in the lungs of cigarette smokers. Am Rev Respir Dis 1983; 128:833-8. 3. Auerbach 0, Garfunkel L, Hammond EC. Relationship of smoking and age to findings in the lung parenchyma: a microscopic study. Chest 1974;65: 29-35. 4. Bridges RB, Kraal H, Huang JT, Chancellor MB. Effects of tobacco smoke on chemotaxis and glucose metabolism of polymorphonuclear leukocytes. Infect Immun 1977; 15:115-23. 5. Kilburn KH, McKenzie N. Leukocyte recruitment to airways by cigarette smoke and particle phase in contrast to cytotoxicity of vapor. Science 1975; 189:634-6. 6. Flint GL, Maxwell KW, Renetti AD. Influence of cigarette smoke on guinea pigs: effects on pulmonary cellsand serum antitrypsin levels.Arch Environ Health 1971; 22:366-9. 7. Rylander R. Pulmonary cell responses to inhaled cigarette smoke. Arch Environ Health 1974; 29:329-33. 8. Rennard SI, Ghafouri M, Thompson AB, et ale Fractional processing of sequential bronchoalveolar lavage to separate bronchial and alveolar samples. Am Rev Respir Dis 1990; 141:208-17. 9. Senior RM, Tegner H, Kuhn C, Ohlsson K, Starcher BC, Pierce JA. The induction of pulmonary emphysema with human leukocyte elastase. Am Rev Respir Dis 1977; 116:469-75. 10. Snider GL, Lucey EC, Christensen TG, et ale Emphysema and bronchial secretory cell metaplasia induced in hamster by human neutrophil products. Am Rev Respir Dis 1984; 129:155-60. 11. Carp H, Janoff A. Potential mediator of inflammation. Phagocyte derived oxidants suppress the elastaseinhibitory capacity of alpha-l-proteinase inhibitor in vitro. J Clin Invest 1980; 66:987-95. 12. Kew RR, Ghebrehiwet B, Janoff A. The fifth component of complement (C5) is necessary for maximal pulmonary leukocytosis in mice chronically exposed to cigarette smoke. Clin Immunol Immunopathol 1987; 43:73-81. 13. Kew RR, Ghebrehiwet B, Janoff A. Cigarette smoke can activate the alternate pathway of complement in vitro by modifying the third component of complement. J Clin Invest 1985;75:1000-7. 14. Kew RR, Ghebrehiwet B, Janoff A. Characterization of the third component of complement (C3) after activation by cigarette smoke. Clin Immunol Immunopathol 1987; 44:248-58. 15. Perricone R, Decarolis C, DeSanctis G, Fontana L. Complement activation by cigarette smoke

767

condensate and tobacco infusion. Arch Environ Health 1983; 38:176-9. 16. Henson PM, McCarthy K, Larsen GL, et ale Complement fragments, alveolarmacrophages, and alveolitis. Am J Pathol 1979; 97:93-110. 17. Gadek JE, Hunninghake GW, Zimmerman RL, Crystal RG. Regulation of the release of alveolar macrophage-derived neutrophil chemotactic factor. Am Rev Respir Dis 1980; 121:723-33. 18. Robbins RA, Justice JM, Rasmussen JK, Russ WD, Thomas KR, Rennard SI. Role of chemotactic factor inactivator in modulating alveolar macrophage-derived neutrophil chemotactic activity. J Lab Clin Med 1987; 109:164-70. 19. Cochrane CG, Miiller-Eberhard HJ. The derivation of two distinct anaphylatoxin activities from the third and fifth components of human complement. 'J Exp Med 1968; 127:371-6. 20. Ward PA, Newman LJ. A neutrophil chemotactic factor from human C5. J Immunol 1978; 129:109-15. 21. Fernandez HN, Henson PM, Otan A, Hugli T. Chemotactic responses to human C3a and C5a anaphylatoxins. I. Evaluation of C3a and C5a leukotaxis in vitro and under simulated in vivo conditions. J Immunol 1978; 120:109-15. 22. Bokisch VA, Muller-Eberhard HJ. Anaphylatoxin inactivator of human plasma: its isolation and characterization as a carboxypeptidase. J Clin Invest 1970; 49:2427-36. 23. Perez HD, Kelley E, Chenoweth D, Elfman F. Identification of the C5a des arg cochemotaxin. Homology with vitamin D-binding protein (groupspecific component globulin). J Clin Invest 1988; 360-3. 24. Kew RR, Webster RO. Gc Globulin (vitamin D-binding protein) enhances the neutrophil chemotactic activity of C5a and C5a des argo J Clin Invest 1988; 82:364-9. 25. Robbins RA, Hamel FG. Chemotactic factor inactivator interaction with Gc-globulin. A mechanism of modulating the chemotactic activity of C5a. J Immunol 1990; 144:2371-6. 26. Berenberg JL, Ward PA. Chemotactic factor inactivator in normal human serum. J Clin Invest 1973; 52:1200-6. 27. Kruetzer DL, Claypool WD, Jones ML, Ward PA. Isolation by hydrophobic chromatography of the chemotactic factor inactivator from human serum. Clin Immunol Immunopathol1979; 12:16276. 28. Robbins RA, Rasmussen JK, Clayton ME, Gossman GL, Kendall TJ, Rennard SI. Antigenic identification of chemotactic factor inactivator in normal human serum and bronchoalveolar lavage fluid. J Lab Clin Med 1987; 110:292-9. 29. Carp H, Janoff A. A possible mechanism of emphysema in smokers. In vitro suppression of serum elastase-inhibitory capacity by fresh cigarette smoke and its prevention by antioxidants. Am Rev Respir Dis 1978; 118:617-21. 30. Kreutzer DL, O'Flaherty JT, Orr FW, Showell HJ, Becker EL, Ward PA. Quantitative comparison of the various biological responses of neutrophils to different active and inactive chemotactic factors. Immunopharmacology 1979; 1:39-47. 31. Martin WJ, Gadek JE, Hunninghake GW, Crystal RG. Oxidant injury of lung parenchyma cells. J Clin Invest 1981; 68:1277-88. 32. Totti N III, McCuster KT, Campbell EJ, Griffin GL, Senior RM. Nicotine is chemotactic for neutrophils and enhances neutrophil responsiveness to chemotactic peptides. Science 1984; 223:169-71. 33. Robbins RA, Russ WD, Rasmussen JK, Clayton MM. Complement activation in the adult respiratory distress syndrome. Am Rev Respir Dis

768 1987; 135:651-8. 34. Bell CY, Haseman A, Spock G, McLennan G, Hook GER. Plasma proteins of the bronchoalveolar surface of the lungs of smokers and nonsmokers. Am Rev Respir Dis 1981; 124:72-9. 35. Metcalf JP, Thompson AB, Rennard SI, Rob-

ROBBINS, GOSSMAN, NELSON, KOYAMA, THOMPSON, AND RENNARD

bins RA, Gc-Globulin functions as a cochemotaxin in the lower respiratory tract (abstract). Chest 1989; 96:131S. 36. Johnson D, Travis J. Structural evidence for methionine at the reactive site of human a-I-proteinase inhibitor. J BioI Chern 1978; 253:7142-4.

37. Johnson P, Travis J. The oxidative inactivation of human u-l-proteinase inhibitor. Further evidence for methionine at the reactive center. J BioI Chern 1979; 254:4022-6.

Inactivation of chemotactic factor inactivator by cigarette smoke. A potential mechanism of modulating neutrophil recruitment to the lung.

Activation of the complement system with generation of the potent neutrophil chemotactic factor C5a has been proposed to play a significant role in th...
682KB Sizes 0 Downloads 0 Views