* Department of Internal Medicine, Justus-Liebig-Universität, Giessen
Department of Rheumatology, Medical University of Lübeck, Germany
Correspondence: F. Grimminger, M.D., Ph.D., Department of Internal Medicine, Justus-Liebig University Giessen, Klinikstrasse 36, D-35392 Giessen, Germany. E-mail: friedrich.grimminger{at}innere.med.uni-giessen.de
 TOP ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Key Words: anti-neutrophil-cytoplasmic antibodies • neutrophil activation • chemotactic peptide • Wegener’s granulomatosis
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
In addition to direct activation of secretory neutrophil functions, sensitizing (priming) of neutrophils for subsequent activation plays an important role in neutrophil-mediated tissue injury. In vitro, when primed with bacterial lipopolysaccharides or proinflammatory cytokines, neutrophils show an enhanced secretory response upon subsequent stimulation with naturally occurring agonists such as bacterial n-formylated peptides or artificial stimuli like calcium ionophores [12 13 14 15 ]. In vivo, if adequately controlled, this enhanced responsiveness of primed neutrophils toward bacterial products can enhance resistance to bacterial infections. However, such up-regulation of PMN responsiveness may also result in progressive tissue destruction under pathophysiological conditions.
Against this background, we investigated whether subthreshold concentrations of anti-PR3-antibodies are capable of sensitizing neutrophils for enhanced responsiveness to proinflammatory agents. Our interest was particularly centered upon bacterial n-formylated peptides because exposure of PMN to these potent agonists is very likely to occur in active WG, with exacerbations of this disease being closely related to infections [16 , 17 ]. In essence, we found that upon preincubation with low doses of anti-PR3 antibodies, neutrophils displayed a dramatically altered response to subsequent stimulation with the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP), but not to the calcium ionophore A23187 or platelet-activating factor (PAF), with marked amplification of the fMLP-induced leukotriene generation and enhanced chemotactic movement toward this peptide. In contrast, elastase secretion and O2- generation were blunted in anti-PR3-pretreated PMN subsequently undergoing fMLP challenge. These anti-PR3-induced alterations of inflammatory neutrophil behavior may contribute to disturbances in host-defense capacity and mechanisms of tissue injury in WG.
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
(TNF-) was from R & D
Systems (Wiesbaden, Germany). Phosphate-buffered saline (PBS) was
obtained from GIBCO Laboratories (Grand Island, NY). The kinetic-OLC
Limulus amebocyte cell lysate test for the detection of
endotoxin was from Chromogenix (Mölndal, Sweden), and
enzyme-linked immunosorbent assay (ELISA) kits for the detection of
PR3-ANCA were from Orgentec (Mainz, Germany). S-2484, a substrate for
neutrophil elastase, was purchased from Kabi-Vitrum (Stockholm,
Sweden). fMLP-lys-FITC, a fluoresceinated peptide that retains fMLP
activity and binding characteristics, was from Peninsula Laboratories
(Belmont, CA). The leukotrienes (LT) LTC4,
LTD4, LTE4, LTB4, 20-OH- and
20-COOH-LTB4, and the synthetic LTA4-methyl
ester were generous gifts from Dr. J. Rokach (Merck Frosst, Toronto,
Ontario, Canada). Additional LTs were graciously supplied by Dr. W.
Bartmann (Hoechst, Frankfurt, Germany). 5-, 8-, 9-, 11-, 12-, And
15-hydroxyeicosatetraenoic acid (HETE), 5(S),12(S)-diHETE, 5,15-diHETE,
and 12-HHT, as well as the non-enzymatic hydrolysis products of
LTA4 (6-trans diasteromeric pairs of
LTB4 and 5,6-diHETEs) were obtained from Paesel (Frankfurt,
Germany). All LTs were checked for purity and quantified
spectrophotometrically before use, as described [19
].
Chromatographic supplies included an analytical high-performance liquid
chromatography (HPLC) column (length x inner diameter = 250
mm x 4 mm; Shandon, Astmoor, UK) filled with ODS-Hypersil
(particle size 3 µm; pore size 100 ;anA; Machery Nagel, Düren,
Germany). HPLC-grade solvents, distilled in glass, were purchased from
Fluka (Heidelberg, Germany). All other biochemicals were obtained from
Merck (Darmstadt, Germany).
Anti-PR3-antibodies
Murine monoclonal antibodies targeting PR3 were prepared by
hybridoma technology, as previously described [18
]. The
clone WGM2 (IgG1) was chosen for further experiments.
F(ab)2 fragments were generated by digestion with pepsin in
0.1 NaAc for 16 h at 37°C. After dialysis against PBS, purity
was checked by sodium dodecyl sulfate-polyacrylamide gel
electrophoresis (SDS-PAGE). ANCA-IgG was isolated from the sera of
three patients with monospecific anti-PR3-positive WG, whereas normal
IgG was purified from the sera of healthy volunteers, through the use
of protein G columns, as described [18
]. Specificity for
PR3 of the human and murine monoclonal antibodies, as well as the
ability of F(ab)2 fragments to recognize PR3 after the
digestion procedure, was assessed in a commercial antigen-specific
ELISA. Endotoxin contamination of the human and murine antibodies was
below 15 pg/mL, as assessed by the kinetic-OLC Limulus
amebocyte lysate (LAL) test.
Isolation of human neutrophils
Neutrophils were isolated from venous blood of healthy donors by
centrifugation over a Ficoll-Paque gradient as previously described
[8
]. In brief, EDTA-anticoagulated blood was layered
over Ficoll-Paque and centrifuged at 400 g for 35 min. After
removal of mononuclear cells, erythrocytes were sedimented in 1%
polyvinyl alcohol. Residual erythrocytes were removed by hypotonic
lysis, cells were washed twice in
Ca2+/Mg2+-free PBS, and finally resuspended in
PBS containing CaCl2 (1 mmol/L) and MgSO4 (1
mmol/L) at 5 x 106 PMN/mL (leukotriene synthesis,
respiratory burst, degranulation) or at 5 x 107
PMN/mL (chemotaxis). Cell purity was >97%, as quantified by flow
cytometry, and cell viability was >96%, as assessed by trypan blue
dye exclusion.
Measurement of leukotrienes
LTs and HETEs were extracted from cell supernatants by
octadecylsilyl solid-phase extraction columns, as described
[8
, 19
]. Reversed-phase HPLC was carried
out on octadecylsilyl columns (Hypersil, 5-µm particles), with a
mobile phase of methanol/water/acetic acid (72:28:0.16, pH 4.9)
[19
]. In addition to the conventional UV detection at
270 nm (LTs) and 237 nm (HETEs), a photodiode array detector (Waters
model 990) was used, which provided full UV spectra (190–600 nm) of
eluting compounds and allowed checking for peak purity and subtraction
of possible coeluting material. All data obtained by the different
analytical procedures were corrected for the respective recoveries and
are given in picomoles per milliliter throughout the experiments.
Recovery was determined by separate recovery experiments using
different quantities of the individual compounds in the appropriate
concentration range. Factors for recovery were further confirmed by
addition of 0.2 µCi of [3H]LTB4 and
5-[3H]-HETE to buffer medium as internal standards in
selected experiments. For quantification of LTs and 5-HETE,
correspondence of values calculated from UV absorbency in two different
chromatographic procedures was required (deviation <10%).
Superoxide anion generation
Neutrophil O2- generation was assessed
as superoxide dismutase-inhibitable reduction of cytochrome
c according to Cohen [20
]. Duplicate reaction
mixtures containing neutrophils (5 x 106/mL) and 75
µM ferricytochrome c were incubated at 37°C in the
presence or absence of 10 µg/mL superoxide dismutase. Incubations
were terminated by centrifugation at 4°C at 1200 g.
O2- release was quantified as nanomoles of
cytochrome c reduction, using an extinction coefficient of
21 mM-1 • cm-1 at 550 nm in a
Uvicon Spectrophotometer.
Release of granular constituents
Elastase was taken as marker for neutrophil degranulation, and
enzyme activity in the cell supernatant was measured by monitoring the
turnover of
L-pyroglutamyl-L-propyl-L-valine-p-nitro-anilide
at 405 nm according to the method described by Kramps et al.
[21
].
Chemotaxis
The chemotactic response of neutrophils to a gradient of fMLP
was measured under agarose according to Nelson et al.
[22
]. In brief, 1.5 mL of a 1% electrophoresis-grade
agarose in medium 199 with 0.5% gelatin, 200 mM glutamine, and 10%
fetal calf serum was adjusted to pH 7.4 and plated on a 35 x
10-mm-diameter disposable plastic Petri dish. Four wells, 2.5 mm in
diameter, were made in the agarose plate with a template punch, and
plugs were removed with suction. Three concentric wells were spaced 5
mm apart to one central well. Neutrophil migration was determined by
placing 10 µL of the chemotactic agent in one (chemotaxin well) and
the appropriate quantity of saline in the remaining two control wells.
Ten microliters of the neutrophil suspension (5 x
107/mL) were added to the central well. The agarose plates
were incubated at 37°C in a 5% CO2 atmosphere for
various time periods and finally fixed with 2.5% glutaraldehyde for
1 h at room temperature. The agarose layers were subsequently
removed from each slide, and the cells were stained with Wright’s
stain. Counting was facilitated by using a slide projector at x40
magnification. The migratory distance was expressed as the chemotactic
index: directed cellular migration toward the chemotaxin well divided
by random cell migration toward saline. All assays were done in
triplicate.
Flow cytometry
For the determination of surface expression of fMLP receptors,
flow cytometry was performed. In brief, isolated neutrophils (5 x
106/mL) were resuspended in PBS and incubated with murine
monoclonal anti-PR3-antibodies (0.1 µg/mL), isotype-matched mouse
control-IgG (0.1 µg/mL), or sham-incubated for various time periods
(1–20 min). Cells were pelleted at 4°C, and resuspended in PBS
containing 0.1% BSA and 0.02% sodium acid. Then, 2 x
105 cells were distributed to each well of flexible
round-bottom microtiter plates, washed, and incubated with 20 µL of
fMLP-lys-FITC (1 µM) for 30 min. After three final washes, cells were
resuspended in PBS and kept on ice until flow cytometric analysis. Flow
cytometry was performed in a FACScan (Becton-Dickinson, Mountain View,
CA) using forward and orthogonal light scatter to select viable cells.
Background fluorescence was quantified by the addition of 1000-fold
excess unlabeled fMLP along with the fluoresceinated analog.
CellQuest® research software (Becton-Dickinson) was used
to analyze the generated data.
Experimental protocols
In the standard protocol, neutrophils were at 5 x
106 PMN/mL and incubated with TNF- (0.5 ng/mL, 15 min)
to induce surface expression of PR3. Then, neutrophils were
preincubated with murine monoclonal anti-PR3-antibodies,
isotype-matched control mouse IgG, F(ab)2 fragments of the
anti-PR3-antibody, human ANCA-IgG, or normal IgG. Monoclonal antibodies
were used at 0.1 µg/mL, and ANCA-IgG or normal IgG was used at 1
µg/mL. After various preincubation periods (2–20 min), neutrophils
were stimulated with fMLP (1 µM), PAF (5 µM), or A23187 (1 µM)
for various time periods (5–20 min). For induction of leukotriene
synthesis, exogenous AA (10 µM) was supplied together with fMLP and
PAF. Reactions were stopped by centrifugation at 4°C (13.000
g, 5 min), and the respective mediators were analyzed in the
cell supernatant.
Statistics
For statistical comparison, one-way analysis of variance (ANOVA)
was performed, followed by Tukey’s honestly significant difference
test when appropriate. A level of P < 0.05 was
considered to be significant.
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
-OH-LTB4 and
-COOH-LTB4, summed up as -Ox-LTB4) in the
cell supernatant (Fig. 1
). In parallel, decay products arising from the non-enzymatic
hydrolysis of LTB4 (6t-LTB4,
6t,12e-LTB4, 5S,6R-DiHETE, 5S,6S-DiHETE; summed up as
LTA4 decay) as well as 5-HETE were liberated from
fMLP-stimulated neutrophils (Fig. 1B
and 1C)
. The kinetics of
liberation differed between these metabolites: while the sum of
LTB4 and its oxidation products, as well as the
LTA4 decay products, displayed progressive accumulation
over the entire incubation period, the release of 5-HETE peaked at 5
min, with subsequent rapid decline. When neutrophils were pretreated
with monoclonal anti-PR3-antibodies (anti-PR3; 0.1 µg/mL) for 5 min,
and subsequently exposed to fMLP and AA, the fMLP-induced liberation of
leukotrienes was markedly amplified. In fact, anti-PR3-pretreated
neutrophils released about 3.5- to 4-fold more 5-LO-mediators than
mono-fMLP-stimulated neutrophils. Preincubation of PMN with equal
concentrations of the anti-PR3 F(ab)2 fragments was
entirely ineffective. Similarly, exposure of PMN to equal
concentrations of an isotype-matched control-IgG had no affect on the
fMLP-induced leukotriene generation. The 5-min anti-PR3 pretreatment
per se, with subsequent mono-exposure to AA, did not induce
any activation of neutrophil leukotriene synthesis under the present
experimental conditions (data not shown).
 View larger version (14K): [in a new window] |
Figure 1. Effect of anti-PR3-antibodies on fMLP-induced leukotriene-synthesis.
Neutrophils (5 x 106/mL) were preincubated with
murine monoclonal anti-PR3-antibodies (anti-PR3), isotype-matched mouse
control IgG (IgG), or F(ab)2 fragments of the
anti-PR3-antibodies [anti-PR3 F(ab)2] for 5 min, or sham
incubation was performed. All antibodies were used at 0.1 µg/mL.
After the preincubation period, neutrophils were stimulated with
fMLP (1 µM) and AA (10 µM) for various time periods (5–20 min),
and cell supernatants were subjected to solid-phase-extraction and
HPLC. 5-LO-metabolites are given in picomoles per milliliter.
Nonenzymatic hydrolysis products of LTA (6t-LTB4,
6t,12e-LTB4, 5S,6R-DiHETE, 5S,6S-DiHETE) are summed up as
LTA4-decay, and products derived from the -oxidation
pathway of LTB4 (20-OH-LTB4 and
20-COOH-LTB4) are indicated as -Ox-LTB4.
Means ± SEM of at least four experiments each are
given. *Significantly different from sham-pretreated, fMLP-stimulated
PMN.
|
 View larger version (15K): [in a new window] |
Figure 2. Time-dependence of the anti-PR3-related priming of leukotriene
synthesis in response to fMLP. Neutrophils (5 x
106/mL) were preincubated for different time periods (0–20
min) with 0.1 µg/mL murine monoclonal anti-PR3-antibodies (anti-PR3),
isotype-matched mouse control IgG (IgG), or sham incubation was
performed. After the preincubation period, neutrophils were stimulated
with fMLP (1 µM) and AA (10 µM) for 10 min. 5-LO-metabolites were
extracted from the cell supernatant and LTB4 release, as
exemplarily depicted, is given in picomoles per milliliter. Means ± SEM of at least four experiments each are given.
*Significantly different from sham-pretreated, fMLP-stimulated PMN.
|
 View larger version (16K): [in a new window] |
Figure 3. Influence of anti-PR3 antibodies on neutrophil migratory behavior
toward fMLP. Neutrophils (5 x 107/mL) were
preincubated with murine monoclonal anti-PR3 antibodies (anti-PR3, 0.1
µg/mL), equal concentrations of an isotype-matched mouse control IgG
(IgGm), F(ab)2 fragments of the anti-PR3
antibodies [anti-PR3 F(ab)2, 0.1 µg/mL), ANCA-IgG from
anti-PR3-postive patients (1 µg/mL; PR3-ANCA) or normal human IgG
(IgGh) or sham-incubated for 5 min. Then, 10 µl of the
neutrophil suspension (50,000 cells) were placed on agarose plates and
exposed to 0.1 µM fMLP (chemotaxin well) or assay buffer (control
well). After various times of incubation (1–4 h) in a 5%
CO2 atmosphere, cells were fixed and stained with Wright’s
stain. Migratory distance is expressed as chemotactic index. Means ± SEM of at least three independent experiments, each with
values performed in triplicate, are given. *Significantly different
from sham-incubated PMN.
|
 View larger version (15K): [in a new window] |
Figure 4. Effect of anti-PR3 antibodies on neutrophil respiratory burst and
degranulation induced by fMLP. PMN (5 x 106 PMN/mL)
were preincubated with murine monoclonal anti-PR3-antibodies (anti-PR3,
0.1 µg/mL), equal concentrations of an isotype-matched mouse control
IgG (IgG), or sham-incubated for 5 min. Then, cells were exposed to
fMLP (1 µM, filled columns) or sham-stimulated (shaded columns) for
an additional 10 min, and secretion products were analyzed. Superoxide
generation is expressed as nmol O2-/5 x
106 PMN (A), and elastase is given in units per liter (B).
Means ± SEM of at least four independent experiments
each are given. *Significantly different from sham-incubated PMN.
|
 View larger version (30K): [in a new window] |
Figure 5. Effect of anti-PR3 antibodies on neutrophil activation with other
agonists than fMLP. Neutrophils (5 x 106/mL) were
pretreated with murine monoclonal anti-PR3 antibodies (anti-PR3, 0.1
µg/mL) or sham pretreated for 5 min, and subsequently exposed to fMLP
(1 µM; filled columns), PAF (5 µM; shaded columns), or A23187 (1
µM; open columns). For induction of leukotriene synthesis, AA (10
µM) was added simultaneously with fMLP or PAF. After an additional 10
min of incubation, secretion products were analyzed in the cell
supernatant (A, LTB4 release; B,
O2- secretion; C, release of elastase) and are
expressed as percentage of sham-pretreated PMN. Means ±
SEM of at least three independent experiments each are
given. *Significantly different from sham-incubated PMN.
|
 View larger version (17K): [in a new window] |
Figure 6. Time course of anti-PR3 preincubation for activation with other
agonists than fMLP. Neutrophils (5 x 106/mL) were
preincubated for different time periods (0–20 min) with 0.1 µg/mL
murine monoclonal anti-PR3-antibodies (anti-PR3), or sham incubation
was performed. After the preincubation period, neutrophils were
stimulated with A23187 (1 µM) or PAF (5 µM) and AA (10 µM) for 10
min. 5-LO-metabolites were extracted from the cell supernatant and
LTB4 release, as exemplarily depicted, is given in
picomoles per milliliter. Means ± SEM of at least
four experiments each are given. No significant differences were
observed between anti-PR3 and sham-pretreated PMN.
|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Indeed, when employed for pre-incubation of neutrophils in
vitro, low doses of anti-PR3 antibodies caused a most impressive
priming of the neutrophil response to the tripeptide fMLP, which
represents a prototype of the family of bacterial
N-formylated peptides [26
, 27
].
Upon pre-exposure of neutrophils to a murine monoclonal anti-PR3
antibody, but not to an isotype-matched control IgG, a massive
amplification of fMLP-induced leukotriene and 5-HETE synthesis was
noted, along with an enhanced chemotactic movement toward the peptide.
This effect was reproduced by ANCA-IgG from patients with monospecific
anti-PR3-positive WG. In contrast, the fMLP-elicited microbicidal
neutrophil functions, namely the respiratory burst and the release of
proteolytic enzymes such as elastase, were diminished upon pretreatment
with anti-PR3 antibodies. This pattern of anti-PR3-related
preactivation of neutrophils thus displays some distinct features
previously not described for "classic" PMN priming agents, such as
LPS, granulocyte-macrophage colony-stimulating factor, or TNF-. (1)
Priming by the autoantibodies was a very rapid event, with a maximum
after 5–7.5 min, which contrasts with the more long-lasting priming
efficacy of LPS and the above-mentioned cytokines
[12
13
14
15
]. (2) Moreover, the anti-PR3-induced priming of
the fMLP response appeared to be transient because it was no longer
observed after incubation periods of 20 min or longer. In general,
neutrophil priming is not an irreversible process. The phenomenon of
reversible neutrophil priming has already been observed upon exposure
to PAF or after hypotonic treatment [28
,
29
], however, the mechanisms underlying the transient
feature of neutrophil priming remain largely to be elucidated. As for
the anti-PR3-induced priming, it is possible that the duration of the
priming signal or its rate of onset are key determinants, or that the
dynamic feature of ligand-receptor interactions [30
] are
responsible for the kinetics of the anti-PR3-induced priming for the
fMLP response. (3) Priming did not occur when PAF or the calcium
ionophore A23187 were used as neutrophil agonists, thus suggesting a
mechanism of pre-activation specific for the chemotactic peptide. To
the best of our knowledge, such selectivity of neutrophil priming for
formylated peptides has hitherto not been described for a circulating
agent. Clearly, the currently described priming phenomenon was not due
to any contamination of the autoantibodies with LPS because
endotoxin-induced priming of PMN presents with completely different
features [12
13
14
15
], and no LPS was detected in the
anti-PR3 preparation when assayed with the kinetic LAL test.
The molecular mechanisms underlying the anti-PR3-induced priming of
neutrophil responsiveness to fMLP largely remain to be elucidated.
Because both the F(ab)2 fragments of the anti-PR3
antibodies and sole targeting of Fc
-receptors (FcRs) by an
isotype-matched control-IgG were noted to be entirely ineffective,
engagement of both PR3 and FcRs is apparently a prerequisite for the
priming phenomenon. This is in line with previous investigations,
addressing direct activation of secretory neutrophil responses by
sufficiently high autoantibody titers [7
,
31
]. One possible explanation for the priming phenomenon
might be an increase in the overall number of neutrophil fMLP receptors
in response to anti-PR3 pretreatment, however, this was excluded in the
FACS studies measuring the binding of FITC-labeled fMLP. Alternatively,
a change in the receptor affinity for N-formylated-peptides
might be operative. The neutrophil fMLP receptor exists in two affinity
states that are regulated dynamically, and changes in receptor affinity
and function are closely related [32
, 33
].
Various agonists have been reported to affect the neutrophil response
toward N-formylated peptides by modifying the fMLP-receptor
affinity in vitro [34
, 35
].
Consistent with the theory that high-affinity receptors mediate
chemotaxis, whereas low-affinity receptors play a role in secretion of
superoxide anion and granule components [36
37
38
], the
anti-PR3-induced alterations of the fMLP response, priming of
chemotactic movement, and leukotriene generation with a decrease in
secretion of O2- and elastase, might reflect a
switch toward predominance of high-affinity fMLP receptors. This is
well in line with previous studies demonstrating that chemotactic
movement and secretory functions induced by fMLP are regulated
independently [36
37
38
]. Furthermore, changes in
characterized post-fMLP receptor signaling pathways
[38
39
40
] might be involved in the anti-PR3-related
priming event. It has already been described that preincubation of
human neutrophils with PR3-ANCA affects fMLP-related generation of
inositolphosphates and membrane translocation of protein kinase C
[41
], which is of interest in view of the role of these
signaling pathways in exocytosis and respiratory burst
[40
, 42
]. Although at first glance
contradicting the previous findings describing a direct activation of
these neutrophil functions by sufficiently high concentrations of
c-ANCA [6
7
8
, 43
], the presently observed
down-regulation of fMLP-induced neutrophil degranulation and
respiratory burst by low concentrations of monoclonal anti-PR3
antibodies might reflect an interference of commonly used signaling
pathways between anti-PR3 and fMLP. In this regard, the recent
demonstration that c-ANCA do possess the capacity to directly activate
PKC [44
] is of particular interest. It may be speculated
that previous activation of PKC by anti-PR3 antibodies may desensitize
this signaling pathway for subsequent engagement with formylated
peptides, as known for well-characterized PKC activators such as
phorbol esters [45
]. Clearly, further studies are
mandatory to clarify the molecular mechanisms underlying the neutrophil
priming by subthreshold doses of anti-PR3 in detail.
In conclusion, low doses of anti-PR3 antibodies, being ineffective per se in causing PMN stimulation, exert a major impact on the human neutrophil responsiveness to fMLP, but not to PAF and A23187. Although generation of leukotrienes as well as the chemotactic response toward the peptide are massively enhanced, respiratory burst and elastase secretion are blunted. On the one hand, such priming may promote neutrophil accumulation at sites of infections via direct (cell migration) and indirect (leukotriene synthesis) mechanisms, and on the other hand, anti-microbicidal capacity may be suppressed. It may be speculated that such an altered response profile of neutrophils under the influence of circulating autoantibodies might contribute to perivascular neutrophil accumulation, persistence of infections, and granuloma formation in Wegener’s granulomatosis.
Received May 15, 2000; revised August 21, 2000; accepted August 22, 2000.
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
RIIa J. Immunol. 153,1271-1280[Abstract], IL-1ß, and IL-2R in ANCA-positive glomerulonephritis Kidney Int 43,682-693[Medline]This article has been cited by other articles:
 |
|
 |
|
  S. M. Uriarte, K. R. McLeish, and R. A. Ward Anti-proteinase 3 antibodies both stimulate and prime human neutrophils Nephrol. Dial. Transplant., April 1, 2009; 24(4): 1150 - 1157. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Hattar, S. van Burck, A. Bickenbach, U. Grandel, U. Maus, J. Lohmeyer, E. Csernok, T. Hartung, W. Seeger, F. Grimminger, et al. Anti-proteinase 3 antibodies (c-ANCA) prime CD14-dependent leukocyte activation J. Leukoc. Biol., October 1, 2005; 78(4): 992 - 1000. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Linden, M. Laan, and G. P. Anderson Neutrophils, interleukin-17A and lung disease Eur. Respir. J., January 1, 2005; 25(1): 159 - 172. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
