 |
||||
| 2007 JASN IMPACT FACTOR 7.111 | HOME AUTHOR INFO EDITORIAL BOARD SUBSCRIBE FEEDBACK ALERTS HELP | |||
| CURRENT ISSUE | ARCHIVES | JASN Express | ONLINE SUBMISSION | |
|
||||
| 2007 JASN IMPACT FACTOR 7.111 | HOME AUTHOR INFO EDITORIAL BOARD SUBSCRIBE FEEDBACK ALERTS HELP | |||
| CURRENT ISSUE | ARCHIVES | JASN Express | ONLINE SUBMISSION | |
*
Department of Medicine, Section of Nephrology, The University of Chicago,
Chicago, Illinois
Department of Medicine, Division of Rheumatology, University of Colorado
Health Sciences Center, Denver, Colorado.
Correspondence to Dr. Richard J. Quigg, Department of Medicine, Section of Nephrology, The University of Chicago, MC 5100, 5841 S. Maryland, Chicago, IL 60637. Phone: 773-702-0757; Fax: 773-702-4816; E-mail: rquigg{at}medicine.uchicago.edu
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Complement activation can proceed via either the classical or alternative pathways. Central to both pathways is the cleavage of C3 and C5, with generation of proinflammatory fragments, such as C3a and C5a, as well as formation of the cell-damaging C5b-9 complex. The complement system is tightly regulated, being constrained in its activation as well as inhibited by a number of regulatory proteins when activation occurs. Beginning with early observations (reviewed in reference 1) that were later extended by Unanue and Dixon (2) and Salant et al. (3), a considerable amount of research has been performed with different experimental glomerular disease models, showing that complement activation is pathogenic (reviewed in reference (4).
In addition to glomerular effects, more recent information has demonstrated that complement activation may lead to tubulointerstitial injury. The circumstances under which this occurs include situations in which complement regulators located on the basal aspects of tubules are inhibited by function-neutralizing antibodies (5,6) and those in which the glomerular barrier to protein passage is impaired (7,8). In the latter setting, complement proteins become accessible to the apical surfaces of the proximal tubule; in such situations, the alternative path-way can be activated (9), because of the lack of C3 convertase regulators at this location (10,11).
A model of renal microvascular injury in which anti-glomerular endothelial cell (GEN) antibodies are injected directly into the renal artery has recently been described (12). Antibody binding to GEN is rapidly followed by complement activation, GEN injury, accumulation of platelets and fibrin in glomeruli, hemolytic anemia, and acute tubular necrosis, features reminiscent of the hemolytic uremic syndrome in human subjects. This renal injury is dependent on formation of the C5b-9 membrane attack complex, inasmuch as C6-deficient PVG rats were protected from disease (13) and inhibition of CD59 (a membrane regulator of C5b-9 formation) exacerbated disease (14).
The mouse protein originally termed p65 (15) was cloned as Crry (CR1-related gene y) (16), and this name has remained in use to signify the mouse and rat proteins (17,18,19). Mouse Crry has decay-accelerating and factor I cofactor activities toward alternative and classical pathway C3 convertases (17,20,21). We have produced transgenic mice in which Crry is expressed as a soluble protein (22). To accomplish this, the active region of mouse Crry was placed under the control of the broadly active and heavy metal-inducible metallothionein I promoter. These Crry transgenic mice exhibit complement-inhibiting levels of Crry in their sera and local production of Crry in various organs, including high levels of expression in renal tubules and glomeruli (22,23). As a result, these mice are protected from glomerular complement activation and injury in the nephrotoxic serum-induced nephritis (NSN) model. In this study, we examined whether these Crry transgenic mice are protected from injury in a model of acute renal failure induced by the intravenous injection of anti-GEN antibodies.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Disease Protocol
Anti-GEN antiserum was raised in a single goat, as described previously
(supplied by Dr. William Couser, University of Washington, Seattle, WA)
(12). IgG was isolated by
protein G affinity chromatography (Pharmacia, Piscataway, NJ) under
endotoxin-free conditions. As assessed with the Limulus amebocyte
lysis assay (Associates of Cape Cod, Falmouth, MA), anti-GEN IgG contained
0.3 EU/ml endotoxin. Anti-GEN IgG was injected intravenously into tail
veins. Animals were allowed to recover from methoxyflurane anesthesia and
disease induction and were then housed in metabolic cages for urine
collection. Twenty h after injection, blood was obtained from the
retro-orbital venous plexus. Thirty h after injection, blood was obtained by
cardiac puncture and urine was collected from the urethral orifice using a
Pasteur pipet, with gentle compression of the bladder. Urine samples collected
in metabolic cages and those collected directly from the urethra were pooled.
Animals were euthanized by cervical dislocation, and renal tissue was
processed for the studies described below.
Two doses of anti-GEN were used in these studies. Eight Crry transgenic animals and seven Crry transgene-negative littermates received 2 mg intravenously. For examination of acute renal C3 deposition, seven transgene-positive and nine transgene-negative animals received injections of 2 mg of anti-GEN, followed by kidney collection 10 or 60 min later. In a separate group of studies designed to examine urinary protein excretion, five transgene-positive and five transgene-negative animals received 0.5 mg intravenously.
NSN Studies
The NSN model was used to determine the influence of proteinuria on acute
renal failure. The advantages of this model for this study are that it is
complement-independent at relatively high doses of NSN antibody and results in
massive proteinuria
(22,24,25).
The basic protocol described above for the anti-GEN disease model was used to
induce NSN. For these studies, transgene-positive or -negative mice received
injections of either 1, 2, or 4 mg of NSN IgG (supplied by Dr. David Salant,
Boston Medical Center).
Localization of Anti-GEN In Vivo
Two approaches were used to determine the localization of anti-GEN in
normal mouse kidneys. The first involved standard indirect immunofluorescence
(IF) microscopic examination of kidney sections from normal CD-1 mice.
Four-micrometer cryostat sections were incubated with anti-GEN IgG, followed
by FITC-conjugated antibody to goat IgG (Cappel, Durham, NC), and were viewed
with a BX-60 IF microscope (Olympus Optical Co., Tokyo, Japan)
(26). Control sections were
similarly processed, except that nonimmune goat IgG was used instead of
anti-GEN IgG.
The second approach involved injection of four normal CD-1 mice with 7.5 mg of anti-GEN IgG, followed by euthanasia 2 or 10 min later (two at each time point). Renal tissue was processed for IF microscopy for goat IgG and mouse C3 as described below.
Measurement of C3 mRNA Levels by Reverse Transcription-PCR
To evaluate whether injection of anti-GEN IgG affected renal transcription
of C3 mRNA
(27,28,29),
the following studies were performed. Twelve normal CD-1 mice received
intravenous injections of 2 mg of anti-GEN IgG. At various times after
injection, animals were euthanized and renal cortical tissue was isolated,
from which RNA was extracted and reverse-transcribed to cDNA using oligo(dT)
primers (19). PCR for mouse C3
was performed using the following primers designed on the basis of the mosue
C3 sequence (GenBank accession number K02782), which span bases 735 to 1234 in
mouse C3 and therefore yield a 500-bp product: forward primer,
5'-GAAT-ACGTGCTGCCCAGTTT-3'; reverse primer,
5'-TGAGTGACCAC-CAGCACTTT-3'. In preliminary studies, reverse
transcription-PCR for C3 in normal mouse renal cortex resulted in linear
accumulation of product from 20 to 35 cycles. For studies evaluating C3 mRNA
accumulation after anti-GEN injection, 23 cycles of PCR were chosen, with the
rationale that anti-GEN might lead to upregulated C3 transcription, which
could be reliably determined using these limiting cycle numbers. As a control
for RNA quantity and integrity, reverse transcription-PCR was also performed
for glyceraldehyde-3-phosphate dehydrogenase (GenBank accession number
M32599), using the following primers, which yield a 195-bp product: forward
primer, 5'-CCATGGAGAAGGCCGGGG-3'; reverse primer,
5'-CAAAGTTGTCATGGATGACC-3'
(30). Twenty cycles of PCR
were found to be in the linear range of product accumulation and were used in
these studies. Photographs of ethidium bromide-stained gels were scanned into
TIFF files and analyzed by Molecular Analyst software (Bio-Rad Laboratories,
Hercules, CA).
Serum and Urine Measurements
Blood urea nitrogen (BUN) and urinary creatinine levels were measured using
a Beckman autoanalyzer (Beckman Coulter, Inc., Fullerton, CA). Total bilirubin
levels in serum were measured using a kit from Sigma Diagnostics (St. Louis,
MO), according to the instructions provided. Urinary excretion of mouse C3 and
albumin was measured using previously described enzyme-linked immunosorbent
assay protocols
(31,32).
Urinary values were normalized to creatinine excretion.
Renal Tissue Processing
Sagittal sections of renal tissue were fixed in buffered formalin.
Five-micrometer sections were stained with periodic acid-Schiff stain and
processed for light microscopic evaluation.
For IF microscopy, tissue was snap-frozen in isopentane on dry ice. Four-micrometer cryostat sections were processed for direct IF microscopy as described previously (26). FITC-conjugated antibodies to mouse C3 and fibrinogen and to goat IgG were obtained from Cappel. The anti-mouse C3 reacts with C3b, iC3b, and C3c but not C3d (31). Rabbit anti-rat C9 was generously provided by Dr. B. Paul Morgan (University of Wales) (33) and was conjugated with FITC (Sigma). This anti-rat C9 antibody cross-reacts with mouse C9, including that in the C5b-9 membrane attack complex, as demonstrated by strongly positive staining of glomeruli from MRL/lpr lupus mice (data not shown). Staining intensities were graded using a previously defined semiquantitative scoring system (32).
An immunohistochemical technique was used to evaluate the numbers of
glomerular neutrophils. Cryostat sections were fixed with 4% paraformaldehyde
and incubated with monoclonal antibody 7/4 (Serotec, Oxford, United Kingdom).
The Vector ABC staining kit (Vector Laboratories, Burlingame, CA) was used
according to the instructions provided by the manufacturer. The numbers of
positively stained cells in 
50 glomeruli/animal were counted. In all
instances when data were being compiled, the observer was blinded to the
origin of the slides.
Statistical Analyses
All data are expressed as mean ± SEM. Comparisons between
transgene-positive and -negative animals were made by t testing. For
comparisons of multiple groups, one-way ANOVA, followed by Tukey's pairwise
comparisons, was used. Correlations among variables were examined by
regression analysis. Minitab software (State College, PA) was used for these
analyses.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Renal Function after Anti-GEN Injection
BUN values were not different between the two groups of animals before
injection of anti-GEN IgG (27.8 ± 1.6 and 32.1 ± 1.5 mg/dl in
transgene-positive and -negative animals, respectively). Twenty h after
anti-GEN injection, BUN levels were significantly higher in transgene-negative
mice (111.0 ± 14.7 mg/dl), compared with transgene-positive mice (48.7
± 11.7 mg/dl), and this persisted until 30 h after injection
(P = 0.007 and 0.012 at 20 and 30 h, respectively).
Figure 2 presents individual
BUN values at the latter time point. As can be observed, five of seven
transgene-negative animals exhibited BUN values of >100 mg/dl, whereas
seven of eight transgene-positive animals exhibited normal BUN values. The
remaining transgene-positive animal exhibited a BUN value of 58.1 mg/dl at the
time of euthanasia.
|
Four of the five transgene-negative animals with BUN values of >100 mg/dl were anuric. For the remaining animal, urinary albumin excretion was 42.0 µg/mg creatinine (normal values, <25 µg/mg creatinine). For all other animals, urinary albumin excretion was only minimally elevated, with average values of 60.2 µg albumin/mg creatinine for the two transgene-negative animals with normal renal function and 73.0 ± 58.2 µg albumin/mg creatinine for the transgene-positive animals. Urinary C3 was not detectable for any animal. Therefore, injection of anti-GEN antibodies produces acute renal failure but only minor apparent alterations in the glomerular perms-electivity barrier to protein passage. Animals bearing the Crry transgene are protected from acute renal failure.
To further investigate the possibility that anti-GEN might induce glomerular proteinuria, five transgene-positive and five transgene-negative animals were given injections of 0.5 mg of anti-GEN, with the rationale that the lower dose would not lead to anuria in the most severely affected animals. Thirty h after injection, BUN values were 32.4 ± 1.4 and 30.4 ± 2.6 mg/dl for the transgene-positive and -negative animals, respectively. Urinary albumin excretion values were 67.9 ± 12.8 and 160.2 ± 82.5 µg albumin/mg creatinine for transgene-positive and -negative animals, respectively. Although they suggested a difference, these values were not statistically different, because of the wide variability. Again, urinary C3 was not detectable for any animal.
Renal Histologic Features
Irrespective of transgene status, animals with BUN values of <100 mg/dl
exhibited normal glomerular (Figure
3a) and tubular (Figure
4a) histologic features. In the five transgene-negative animals
with renal failure, as defined by BUN values of >100 mg/dl, there were
varying degrees of focal glomerular capillary collapse
(Figure 3b) and acute tubular
necrosis (Figure 4b).
Occasional polymorphonuclear leukocytes were observed in the glomeruli of most
animals, but marked exudative changes were not apparent (further quantified
below). Interstitial infiltrates were not present.
|
|
IF Findings
Thirty h after injection, goat IgG persisted in the glomeruli of all
animals (data not shown). Although there was some variability in staining
intensity among animals, there was no statistical difference between
transgene-negative and -positive animals
(Table 1).
|
Coincident with the presence of goat IgG in glomeruli, modest amounts of mouse C3 were apparent in glomeruli. Interestingly, there was no difference between transgene-positive and -negative animals (Table 1 and Figure 5). To examine complement activation occurring acutely after anti-GEN injection, animals were euthanized 10 or 60 min after antibody injection. In transgene-negative animals, there was segmental, finely granular, glomerular capillary wall staining for C3 in two of four and five of five animals at 10 and 60 min, respectively, which was not present in any of the seven transgene-positive animals at these two time points.
|
In contrast to glomerular C3 staining, in transgene-negative animals there was prominent tubular staining for C3 both in a basal distribution (Figure 6b) and in tubular cells (including those that may have been sloughed into the tubular lumina) (Figure 6c), which was significantly less than that observed for transgene-positive animals (Table 1 and Figure 6a). Interestingly, the tubular cell staining for C3 was strongly correlated with BUN values (r = 0.83, P < 0.001), suggesting a causal role in renal failure.
|
IF staining for C9 protein was also examined. In contrast to C3, C9 was not identified to an appreciable extent in glomeruli, nor was it present in the basal aspects of tubules (data not shown). However, tubular cells stained for C9 to an extent equivalent to that for C3 (Table 1), and the two values were thus strongly correlated (r = 0.79, P < 0.001). As with C3, C9 staining was correlated with BUN levels (r = 0.56, P = 0.037). These data further suggest that complement activation, leading to the generation of C5b-9 on tubules, is pathogenic in this model.
Renal Cortical C3 mRNA Levels
After injection of anti-GEN IgG, C3 mRNA levels in renal cortex were
evaluated, with the rational that de novo synthesis of C3 may occur
locally after injury in this model. However, there was no significant
difference with time in C3 mRNA levels
(Figure 7). These results
indicate that C3 transcription is not increased in the renal cortex in this
model. In fact, there was a slight decrease in C3 mRNA levels at the later
time points (20 h), which possibly could be ascribed to tubular cell
damage.
|
Neutrophil Accumulation
Thirty h after injection of anti-GEN IgG, there was a modest accumulation
of neutrophils in glomeruli. However, there was no difference between
transgene-positive and -negative animals (0.35 ± 0.17 and 0.33 ±
0.09 neutrophils/glomerulus, respectively).
Thrombotic Microangiopathy
In rats given anti-GEN directly in the renal artery, a model of the
hemolytic uremic syndrome develops, characterized by hemolysis and glomerular
fibrin accumulation (12). In
this study, however, there was no specific glomerular accumulation of fibrin,
as assessed using IF microscopy (data not shown). Furthermore, total bilirubin
levels in sera 30 h after disease onset were not elevated and were not
different between the groups (0.67 ± 0.39 and 0.58 ± 0.17 mg/dl
in transgene-positive and -negative animals, respectively). Therefore, this
model in mice seems to be distinct from that induced in rats.
NSN Studies
In mice given relatively high doses of NSN IgG, the resultant glomerular
injury is complement-independent and leads to massive proteinuria
(22,24,25).
Comparable amounts of NSN antibody were used in these studies. As demonstrated
in Figure 8, acute renal
failure developed in all animals and was related to the dose of NSN antibody
used. The resultant albuminuria was marked, with values of >10,000 µg
albumin/mg creatinine for every animal, and values were positively correlated
with BUN values (r = 0.69, P < 0.001). In contrast to
studies in animals with acute renal failure induced by anti-GEN, no animal in
these studies was anuric. Furthermore, instead of diffuse tubular cell
staining for mouse C3, there was punctate staining for mouse C3 in apical
portions of tubular cells, consistent with these being endocytic vacuoles
(Figure 9) (34).
|
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The finding that Crry transgenic animals were protected from acute renal failure implicates the complement system as being pathogenic in this model, as is the case in the rat model (13). The possible sites of complement activation within the kidney in this model include the glomerular, tubular, and vascular compartments. On the basis of our previous results with the NSN model, in which glomerular complement activation was eliminated in 17 of 20 Crry transgenic mice (22), it was logical to assume that a similar degree of complement inhibition would occur in this model. This was true early in the course of this disease model, but by 30 h the difference in C3 immunostaining was no longer present. Crry acts as a factor I cofactor to cleave C3b into the inactive fragment, iC3b (21). Because this is also recognized by the anti-C3 antiserum used for IF (31), a plausible explanation for these findings is that anti-GEN did activate complement but in Crry transgenic animals the C3b generated was inactivated by Crry but remained covalently bound in glomeruli. The next logical site of antibody binding and complement activation is within the renal vasculature, as observed with anti-GEN injection in rats (12), as well as with injection of neutralizing anti-Crry antibodies (5). However, we could not identify specific antibody binding or C3 deposition in the renal microvasculature, either acutely or 30 h after injection of anti-GEN antibodies.
Tubular C3 staining was significantly greater in wild-type mice, compared with Crry transgenic mice; this finding was true for C3 staining in both basal (presumably the tubular basement membrane) and cellular distributions. This basal distribution of C3 is a normal finding in mice, as is focal and segmental staining of Bowman's capsule, albeit in lesser amounts than present in this study in experimental transgene-negative mice. This staining is specific for the C3 protein, because it is not present in C3-deficient mice (32). The basal staining for C3 was significantly increased in transgene-negative controls. However, the fact that this staining bore no relationship to BUN levels (r = 0.32) makes it less likely that basal complement activation is responsible for the acute renal failure in this model.
In contrast to the findings presented above, the high degree of correlation between tubular cell staining for C3 and BUN levels (r = 0.83) suggests that C3 deposition is at least a marker for renal failure and may be causally related. This finding is despite the fact that the defect in glomerular perm-selectivity was not marked, as indicated by mild albuminuria and the absence of detectable urinary C3. To further examine the relationship between proteinuria and acute renal failure, we used the NSN model. Animals developed acute renal failure, the magnitude of which was proportional to the amount of NSN antibody injected and the degree of albuminuria. Rather than the diffuse tubular cell staining for C3 observed in anti-GEN-treated animals, mice with NSN exhibited evidence of endocytic vacuoles (34), consistent with active reabsorption of C3 leaking across the glomerular capillary wall. Whether tubular complement activation contributes to pathogenesis in the NSN model but overwhelms protection by the soluble Crry protein is not clear.
Taken together, the anti-GEN model developed here must include the following initial glomerular events: binding of injected antibodies to glomerular endothelium, complement activation, modest impairment of permselectivity, and accumulation of neutrophils. Complement activation on tubules then ensues, leading to acute renal failure. Potential mechanisms for this tubular complement activation include ischemia-reperfusion injury (35,36,37) related to the glomerular damage; complement activation in peritubular capillaries by goat anti-GEN or natural murine anti-endothelium antibodies (37); and the appearance of complement proteins in the urinary space, either derived from plasma by glomerular filtration or directly produced by tubules (27,29). In this study, we have provided evidence that antibody binding and complement activation do not occur in peritubular capillaries, glomerular filtration of C3 is not apparent, and renal expression of C3 mRNA is not increased. Therefore, of these mechanisms, primary glomerular damage leading to ischemic and possibly reperfusion-related injury of downstream tubular segments is the most plausible explanation that incorporates the available data. A similar degree of widespread tubular complement activation has been noted in human kidney specimens after renal infarction (38). The means by which complement is activated in ischemia-reperfusion injury remain to be fully defined. Animals that express soluble Crry in tubules are resistant to tubular complement activation and the resultant acute renal failure.
Acute renal failure in the setting of well defined complement activation was previously observed in two situations. The first is the anti-GEN model in rats. In this model, prior complement depletion with cobra venom factor reduced BUN levels from 186.1 to 28.3 mg/dl (13). The second is acute renal failure in rats in which massive glomerular proteinuria was induced with puromycin aminonucleoside (7). Complement inhibition with soluble recombinant human CR1 reduced the pathologic indices of tubular damage but did not affect the markedly reduced GFR. Here we show complete prevention of acute renal failure in Crry transgenic mice.
There are distinct advantages to the use of Crry transgenic mice, as in these studies. The first is that complement inhibition is continuous and relatively constant. The second is related to the local production of the complement inhibitor Crry. In the design of these Crry transgenic mice, we focused on the kidney. In the absence of a suitable promoter to yield kidney-specific production of Crry, we chose the metallothionein I promoter; although this promoter is widely active, it exhibits relatively high renals expression (39). We think that high tubular expression of the Crry transgene, which leads to significant release of biologically active Crry in tubular lumina (22), constitutes a significant benefit, particularly given the normal absence of C3 convertase regulators at this site (7,8,9,10). Success in these studies supports the design of similar genetic strategies for human subjects, such as the use of kidney-specific promoters to direct the production of complement inhibitors (40).
|
Acknowledgments |
|---|
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
enhances the biosynthesis of complement C3 and factor B
by human kidney proximal tubular epithelial cells in vitro. Mol
Immunol 33:847
-854, 1996[Medline]
This article has been cited by other articles:
 |
|
 |
|
  J. J. Alexander, A. Jacob, L. Bao, R. L. Macdonald, and R. J. Quigg Complement-Dependent Apoptosis and Inflammatory Gene Changes in Murine Lupus Cerebritis J. Immunol., December 15, 2005; 175(12): 8312 - 8319. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. L. Servin Pathogenesis of Afa/Dr Diffusely Adhering Escherichia coli Clin. Microbiol. Rev., April 1, 2005; 18(2): 264 - 292. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. J. Quigg Complement and the Kidney J. Immunol., October 1, 2003; 171(7): 3319 - 3324. [Full Text] [PDF]
|
|
|
|
 |
|
 M. Nangaku Complement Regulatory Proteins: Are They Important in Disease? J. Am. Soc. Nephrol., September 1, 2003; 14(9): 2411 - 2413. [Full Text] [PDF]
|
|
|
|
|
|
L. Bao, M. Haas, S. A. Boackle, D. M. Kraus, P. N. Cunningham, P. Park, J. J. Alexander, R. K. Anderson, K. Culhane, V. M. Holers, et al. Transgenic Expression of a Soluble Complement Inhibitor Protects Against Renal Disease and Promotes Survival in MRL/lpr Mice J. Immunol., April 1, 2002; 168(7): 3601 - 3607. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Sogabe, M. Nangaku, Y. Ishibashi, T. Wada, T. Fujita, X. Sun, T. Miwa, M. P. Madaio, and W.-C. Song Increased Susceptibility of Decay-Accelerating Factor Deficient Mice to Anti-Glomerular Basement Membrane Glomerulonephritis J. Immunol., September 1, 2001; 167(5): 2791 - 2797. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. PARK, M. HAAS, P. N. CUNNINGHAM, J. J. ALEXANDER, L. BAO, J. M. GUTHRIDGE, D. M. KRAUS, V. M. HOLERS, and R. J. QUIGG Inhibiting the Complement System Does Not Reduce Injury in Renal Ischemia Reperfusion J. Am. Soc. Nephrol., July 1, 2001; 12(7): 1383 - 1390. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
HOME
CURRENT ISSUE
ARCHIVES
JASN Express
ONLINE SUBMISSION
AUTHOR INFO
EDITORIAL BOARD SUBSCRIBE FEEDBACK ALERTS HELP |
) and littermates without the
transgene ([UNK]); BUN values were measured 20 h later. Individual data points
are shown, with the average for each group being depicted as a horizontal
line.