Expression of the Chemokine Monocyte Chemoattractant Protein-1 and Its Receptor Chemokine Receptor 2 in Human Crescentic Glomerulonephritis
STEPHAN SEGERER*,
YAN CUI*,
KELLY L. HUDKINS*,
TRACY GOODPASTER*,
FRANK EITNER,
MATTHIAS MACK,
DETLEF SCHLÖNDORFF and
CHARLES E. ALPERS*
*Department of Pathology, University of Washington, Seattle,
Washington Medizinische Klinik II, Klinikum der Rheinisch-Westfaelischen Technischen
Hoshschule, Aachen, Germany Medizinische Poliklinik, Klinikum der Universitaet, Munich,
Germany.
Correspondence to Dr. Stephan Segerer, University of Washington, Department of
Pathology, C. Alpers Laboratory, Room C415, Medical Center, Box 357470, 1959
NE Pacific Street, Seattle, WA 98195. Phone: 206-543-5616; Fax: 206-543-3644;
Email:
ssegerer{at}u.washington.edu
Abstract. Crescents are morphologic manifestations of severe
glomerularinjury. Several chemokines and their receptors have been
demonstratedto be involved in animal models of crescentic glomerulonephritis
(cGN)and are potential targets for therapeutic interventions. Therefore,the
expression of monocyte chemoattractant protein-1 (MCP-1),its receptor
chemokine receptor 2B (CCR2B), and CCR5 in humancGN was studied. MCP-1 and
CCR2B mRNA expression was evaluated,by in situ hybridization, in
serial sections of 23 renal biopsiesfrom patients with cGN. T cells,
macrophages, and CCR5-expressingcells were examined by immunohistochemical
analysis. MCP-1 mRNAwas expressed by cells in crescents, parietal epithelium,
andtubular epithelium, as well as by infiltrating leukocytes inthe
tubulointerstitium. The expression of CCR2B mRNA was observedin cells in
glomeruli and crescents and in infiltrating leukocytesin the
tubulointerstitium. CCR2B mRNA expression could not beclearly localized to
intrinsic renal cells; evidence that mostof the CCR2B-expressing cells were
leukocytes is provided. CD3-positiveT cells formed the major part of the
interstitial cell infiltratesbut were rare within the glomerular tufts.
CD68-positive macrophagesconstituted a major population of infiltrating cells
in crescentsand contributed significantly to the interstitial infiltrates.
Thenumber of glomerular macrophages was associated with the numberof MCP-1-
and CCR2B-positive glomerular cells. Expression ofCCR2B was significantly
correlated with interstitial CD3-positiveT cells. CCR5 expression was
restricted to infiltrating leukocytesand was correlated quantitatively and by
localization with interstitialCD3-positive T cells and CD68-positive
macrophages. These firstmorphologic data on the distribution of CCR2-positive
cellsin human cGN suggest differential effects of chemokines andtheir
receptors on the distribution of infiltrating leukocytesin different
compartments of the kidney.
Crescents, defined as two or more layers of cells partiallyor completely
filling Bowman's space, are morphologic expressionsof severe inflammatory
glomerular injury and indicators of poorprognoses
(1). Proliferating parietal
epithelium and macrophagesare thought to be major contributors to the cells
in crescents,but T cells and fibroblasts are also involved
(2,3).
The cellcomposition and factors that lead to the formation and resolutionof
crescents are currently under intensive investigation
(4,5,6).
Accumulatingdata in animal models and human subjects point to the importance
ofa cell-mediated immune response in crescentic glomerulonephritis(cGN) (for
review, see reference 7). The
process of leukocyteextravasation from the circulation to the site of
inflammationinvolves a cascade of interactions between soluble factors and
surfacemolecules expressed by leukocytes and endothelial cells
(8).The importance of
chemokines and their receptors at multiplestages in this process has been
demonstrated in the past decade.Chemokines are a superfamily of small
chemotactic cytokinesthat specifically attract subsets of leukocytes (for
review,see references
9,10,11,12).
Chemokines function via G protein-coupledreceptors, which usually bind more
than one chemokine of thesame subgroup. The CC chemokines monocyte
chemoattractant protein-1(MCP-1), which binds to the CC chemokine receptor 2
(CCR2),and regulated upon activation, normal T cell-expressed and secreted
(RANTES),which binds to CCR5, CCR3, and CCR1, have been most extensively
studiedin rodent models of renal diseases (for review, see reference
9).In vitro data
indicate that the chemokine receptors CCR2 andCCR5 are both expressed by at
least some subsets of T cellsand macrophages, which are the main cell types
comprising theinterstitial infiltrates in inflammatory renal diseases andare
of major prognostic importance
(13,14).
The distributionof CCR2-positive cells in human renal diseases is still
unknown.We previously described the expression of CCR5 by infiltratingcells
in various "noncrescentic" renal diseases and demonstratedthat
these cells (mainly T cells) form a major part of the interstitial
inflammatorycell infiltrates commonly observed in renal diseases
(15). Thecharacterization of
chemokine receptor-expressing cells, correspondingto specific morphologically
defined processes in the evolutionof the diseases, can provide new insights
into the pathogenesisof cGN and is a prerequisite for studies with
receptor-blockingagents.
Tissue Samples
A total of 23 renal biopsies from patients with cGN were studied.Included
cases were from biopsies examined during the periodfrom 1996 to 1999 in the
Department of Pathology, Universityof Washington (Seattle, WA). All
accessioned cases for whichsufficient material was available for multiple
additional immunohistochemicalstudies after completion of routine diagnostic
evaluations wereincluded. No further selection criteria were applied. Patient
sampleswere obtained under the general consent to have a renal biopsy
performedand/or accessioned at the University of Washington. Under these
conditions,which were approved by the University of Washington internal
reviewboard for human subjects, no patient identifiers may be usedin studies
involving the biopsy tissue; therefore, correlativefunctional parameters
could not be obtained for this study.The specimens were fixed in 10%
phosphate-buffered formalin,processed, embedded in paraffin, and sectioned at
4 µm,using conventional techniques. Chemokine receptor-overexpressingcell
lines were used as positive controls for in situ hybridizationand
Northern blotting. HOS cells overexpressing CCR1, CCR3,CCR4, CCR5, and CXC
chemokine receptor 4 (CXCR4) were obtainedthrough the AIDS Research and
Reference Reagent Program (Divisionof AIDS, National Institute of Allergy and
Infectious Diseases,National Institutes of Health), to which they were
originallyprovided by Dr. N. R. Landau (Salk Institute for Biological
Studies,La Jolla, CA) CXC chemokine receptor 4
(CXCR4)(16).
CCR2B-overexpressingcells were kindly provided by Dr. I. F. Charo, Gladstone
Instituteof Cardiovascular Disease (San Francisco, CA).
Molecular Probes
The establishment of the MCP-1 probe was previously describedin detail
(17). The cDNA for CCR2B was
kindly provided by Dr.I. F. Charo
(18). For nonradioactive
Northern blotting, thelinearized CCR2B plasmid was transcribed into a
digoxigenin-labeledriboprobe using digoxigenin-labeled UTP (Boehringer
MannheimBiochemicals, Indianapolis, IN). The 28S probe was previously
described
(19,20).
In Situ Hybridization In situ hybridization was performed as described previously
(19,20).
Slideswere deparaffinized in xylene and rehydrated in graded ethanols.After
rinsing with 0.5x SSC (1x SSC is 150 mM NaCl, 15 mM sodium
citrate,pH 7.0), the tissue was digested for 30 min at 37°C with
proteinaseK type XI (5 µg/ml; Sigma Chemical Co., St. Louis, MO)in
proteinase K buffer (500 mM NaCl, 10 mM Tris, pH 8.0). Slideswere rinsed
three times with 0.5x SSC, dehydrated, and air-dried,followed by a 2-h
incubation in 100 µl of prehybridizationbuffer (0.3 M NaCl, 20 mM Tris, pH
8.0, 5 mM ethylenediaminetetraacetate,1x Denhardt's solution, 10%
dextran sulfate, 10 mM dithiothreitol)at 50°C. Hybridization was
performed at 50°C with 50µl of prehybridization buffer containing
500,000 to 700,000cpm of 35S-labeled riboprobe/slide. After
overnight hybridization,slides were washed three times with 2x SSC and
than treatedwith RNase A type IIA (20 µg/ml; Sigma) for 30 min at
37°C.This treatment was followed by three 30-s washes at room temperature
with2x SSC and three high-stringency, 40-min washes at room temperature
with0.1x SSC (containing 0.5% Tween) at 50°C. After three washes
with2x SSC, slides were dehydrated in graded ethanols containing0.3 M
ammonium acetate and were air-dried. After being dippedin NTB2 nuclear
emulsion (Kodak, Rochester, NY), slides wereexposed in the dark at 4°C
for 2 to 8 wk. After development,slides were counterstained with hematoxylin
and eosin, dehydrated,and coverslipped with Histomount (National Diagnostics,
Atlanta,GA).
Northern Analysis
Total RNA was isolated from chemokine receptor-overexpressingHOS cells
using the TOTALLY RNA total RNA isolation kit (Ambion,Austin, TX), according
to the instructions provided by the manufacturer.RNA samples containing 10
µg/lane were separated by electrophoresesthrough a 1% agarose-formaldehyde
gel. The RNA was transferredto a nylon membrane (BrightStar-Plus; Ambion)
using the VacuGeneXL vacuum blotting system (Pharmacia Biotech, Piscataway,
NJ)and was cross-linked to the membrane with ultraviolet light.The part with
the size maker was removed, rinsed with 5% aceticacid for 15 min, and stained
with methylene blue (0.5 M sodiumacetate, pH 5.2, 0.04% methylene blue). The
membrane was placedin prewarmed hybridization buffer (NorthernMax; Ambion)
for2 h at 65°C, followed by overnight hybridization at 65°Cwith the
digoxigenin-labeled riboprobe at a concentration of10 ng/ml. The membrane was
washed twice for 5 min at room temperaturewith 2x SSC containing 0.1%
sodium dodecyl sulfate, followedby two highstringency washes for 15 min at
65°C with 0.1xSSC containing 0.1% sodium dodecyl sulfate. The
BrightStar BioDetectnonisotopic detection system (Ambion) was used, according
tothe instructions provided by the manufacturer, for developmentof the
signal.
Immunohistochemical Analysis
The MC5 antibody against human CCR5 and its specific use for
paraffin-embeddedtissue were previously described
(15). Antibodies to
CD3-positiveT cells (rabbit anti-human CD3 antibody A0452; Dako, Carpinteria,
CA)and CD68-positive macrophages (mouse monoclonal anti-human CD68antibody,
clone PG-M1; Dako) were used for immunohistochemicalanalyses, similar to
previously described protocols
(15,21).
Antigenretrieval was performed on deparaffinized and rehydrated slides,by
steam-heating in antigen-unmasking solution (Vector, Burlingame,CA).
Endogenous peroxidase was blocked by incubation with 3%hydrogen peroxide. The
avidin/biotin blocking kit (Vector) wasused to block endogenous biotin. The
primary antibodies werediluted in phosphate-buffered saline containing 1%
bovine serumalbumin (Sigma) and were applied for 1 h. After subsequent washes
withphosphate-buffered saline, the tissues were incubated with biotinylated
secondaryantibodies for 30 min (goat anti-rabbit Ig and horse anti-mouseIg;
Vector). The ABC-Elite reagent (Vector) was used for signalamplification.
Diaminobenzidine was applied as a chromogen,and slides were counterstained
with methyl green, dehydrated,and coverslipped.
Ten high-power fields (magnification, x400), encompassing anarea of
0.189 mm2 each, were evaluated for each biopsy. Individualpositive
cells were counted in the interstitium and glomeruli.A cell was considered
positive by in situ hybridization whenmore than four silver grains
were concentrated above or immediatelyadjacent to a nuclear profile. Because
of the somewhat punctate,cytoplasmic staining pattern of CD68-positive cells,
quantitativeevaluation was difficult, especially in the tubulointerstitium.
Tominimize this problem, we counted the color reaction productas indicating
a positive cell only when it was associated witha nucleus. This approach
introduces the potential problem ofunderestimation of the number of
interstitial macrophages.
Statistical Analyses
Results are given as means ± SEM. The Wilcoxon test wasused for the
comparison of mean values. Evaluations of the correlationswere performed
using the Spearman correlation coefficient. P< 0.05 was
considered statistically significant.
CD3-Positive T Cells Form a Major Part of Interstitial Infiltrates
but Are Rare in Glomeruli
A prominent feature of all cases was infiltration of the tubulointerstitium
bymononuclear leukocytes (Figure 1, A and
B). The mononuclearcell infiltrates were often accentuated around
glomeruli (Figure 1A).CD3-positive T cells formed a major part of the interstitialinfiltrates
(Figure 1A;
Table 1). Infiltration of the
tubularepithelium by CD3-positive T cells was regularly observed and
consistedof up to four cells/tubular cross-section. Small, nodular,
interstitialinfiltrates of mononuclear leukocytes consisted mainly of T
cells.
Figure 1. Distribution of T cells, macrophages, and chemokine receptor 5
(CCR5)-positive cells. (A) Immunohistochemical analysis of CD3-positive T
cells, showing interstitial CD3-positive T cells (immunohistochemical signal
indicated by the black product) accentuated in the periglomerular area but no
positive cells in the glomerulus. Original magnification, x200. (B)
Immunohistochemical analysis of CD68-positive macrophages. In addition to the
interstitial infiltrate, several macrophages are evident within the glomerulus
(arrowheads). Original magnification, x200. (C) Immunohistochemical
analysis of CCR5-positive cells. The cell distribution mirrors that of
CD3-positive T cells, with no positive cells within the glomerulus. Original
magnification, x200.
Table 1. Mean ± SEM of positive cell numbers per high-power field
In contrast to the large number of tubulointerstitial CD3-positiveT cells,
the number of T cells observed within glomeruli wassmall. Only 1.6% of the
total number of T cells were localizedin glomerular tufts or within
crescents. Bowman's capsule, whichnormally separates the glomerular space
from the interstitium,was usually destroyed in the subset of crescents with
largernumbers of CD3-positive T cells.
CD68-Positive Macrophages Are the Main Infiltrating Cells in
Glomeruli and Crescents
CD68-positive macrophages were common in the tubulointerstitialinfiltrates
(Figure 1B). As discussed
above, the number of interstitialmacrophages might be biased by the staining
pattern; therefore,the absolute numbers of T cells and macrophages cannot
reliablybe compared. Although the distributions of T cells and macrophagesin
the interstitium were often similar, we observed no numericalcorrelation
between the mean number of CD3-positive T cellsand the mean number of
CD68-positive macrophages (data not shown).CD68-positive macrophages were
commonly observed within thetubular lumina.
In addition to comprising a large percentage of the interstitial
infiltrates,CD68-positive macrophages were the main infiltrating cell typein
crescents and glomerular tufts (Figure
1B). The glomerulararea contained a mean of 7.0 CD68-positive
macrophages/high-powerfield and 17.5% of the total number of macrophages. In
comparison,only 1.6% of the total number of CD3-positive T cells were
observedin glomeruli. Seventy-five percent of glomeruli with crescents
containedthree or more macrophages (compared with 22% with three or more
CD3-positiveT cells) (Table
2).
Table 2. Percentage of glomeruli with crescents including three or more positive
cells
Tubular Epithelial Cells, Interstitial Infiltrating Cells, and Cells
in the Glomerular Tufts and in Crescents Express MCP-1
Representative examples from biopsies hybridized with the MCP-1sense and
antisense riboprobes are presented in
Figure 2, A and B.Only the
antisense riboprobe revealed focal hybridization,with the large number of
silver grains signaling a positivereaction
(Figure 2B). Sites of MCP-1
expression were associatedwith mononuclear cell infiltration in both the
glomeruli andtubulointerstitium. In contrast, MCP-1 mRNA expression was
rarelyobserved in areas of well preserved renal architecture.
Figure 2. Monocyte chemoattractant protein-1 (MCP-1) mRNA expression in the
tubulointerstitium. (A) In situ hybridization using a MCP-1 sense
riboprobe, demonstrating a small number of nonspecifically deposited silver
grains. Original magnification, x1000. (B) In situ
hybridization using a MCP-1 antisense riboprobe, demonstrating MCP-1 mRNA
expression by tubular epithelial cells (in situ signal indicated by
the deposition of silver grains). Original magnification, x1000. (C and
D) In situ hybridization using a MCP-1 antisense riboprobe,
demonstrating MCP-1 mRNA expression by infiltrating interstitial cells.
Original magnification: x400 in C; x1000 in D.
In the tubulointerstitium, MCP-1 mRNA was expressed by tubularepithelium
(morphologically, primarily of distal tubules) andinfiltrating mononuclear
cells (Figure 2, B to D). MCP-1
mRNAremained detectable in atrophic tubules. MCP-1 mRNA-expressingcells,
similar to CD68-positive macrophages, were sometimesobserved within tubular
lumina. MCP-1-positive cells were rarein areas of marked nodular accumulation
of mononuclear leukocytes.Biopsies containing larger numbers of MCP-1
mRNA-positive cellsdemonstrated significantly larger numbers of infiltrating
CD3-positiveT cells (Figure
3).
Figure 3. Comparison of the mean numbers of CD3-positive T cells in two groups, with
smaller (left) or larger (right) numbers of MCP-1-expressing cells (P
< 0.05). Groups were formed according to the median of the mean number of
MCP-1-positive cells.
MCP-1 mRNA was expressed by cells in the glomerular tuft, bycells in
crescents, and by parietal epithelial cells
(Figure 4, B and C).Sixty-six
percent of glomeruli with crescents containedthree or more MCP-1-positive
cells and, overall, 12% of MCP-1-positivecells were observed in glomeruli.
This distribution mirroredthe distribution of CD68-positive macrophages.
Biopsies containinglarge numbers of glomerular MCP-1-positive cells displayed
correspondinglylarger numbers of CD68-positive cells within the glomeruli,
butthe difference did not reach statistical significance
(Figure 5).
Figure 4. Expression of MCP-1 mRNA in cells of the glomerular tuft, cells of
crescents, and Bowman's capsule epithelium. (A) Glomerulus with a cellular
crescent (silver-methenamine staining). Original magnification, x400.
(B) In situ hybridization using a MCP-1 antisense riboprobe,
demonstrating MCP-1 mRNA expression in cells of the remaining tuft and cells
of the cellular crescent surrounding the tuft. Original magnification,
x400. (C) In situ hybridization using a MCP-1 antisense
riboprobe, demonstrating MCP-1 mRNA expression by parietal epithelial cells
and cells within the glomerular tuft. Original magnification, x1000.
Figure 5. Comparison of the mean numbers of glomerular CD68-positive macrophages in
two groups, with smaller (left) or larger (right) numbers of glomerular
MCP-1-expressing cells (NS). Groups were formed according to the median of the
mean number of glomerular MCP-1-positive cells.
CCR2B mRNA Is Commonly Expressed by Infiltrating Cells in Glomeruli
and in the Tubulointerstitium
The specificity of the CCR2B riboprobe was demonstrated by Northern
blotting(Figure 6) and by
in situ hybridization of transfected HOS cellsoverexpressing the
chemokine receptors CXCR4 and CCR1 to CCR5.A single band with the expected
size for CCR2B was demonstratedonly in the CCR2B-overexpressing cell line by
Northern blotting.No cross-hybridization was detected in RNA isolated from
cellsexpressing CCR1, CCR3, CCR4, CCR5, or CXCR4
(Figure 6). Usingthe CCR2B
antisense riboprobe for in situ hybridization, significantdeposition
of silver grains was detected only in pellets ofCCR2B-overexpressing HOS
cells (data not shown). There was nosignificant deposition of silver grains
in tissue hybridizedwith the sense riboprobe, as illustrated in
Figure 7.
Figure 6. (A) Northern blot hybridized with a digoxigenin-labeled antisense probe
directed against CCR2B, demonstrating a single band (3.5 kb) in
CCR2B-overexpressing cells. (B) Hybridization with a 28S probe, demonstrating
similar amounts of RNA in each lane.
Figure 7. Distribution of T cells and CCR2B mRNA-positive cells. (A and B) In
situ hybridization using a CCR2B sense riboprobe, demonstrating a small
amount of nonspecific deposition of silver grains. Original magnification:
x400 in A; x1000 in B. (C and D) In situ hybridization
using a CCR2B antisense riboprobe, demonstrating CCR2B mRNA expression in
infiltrating cells (replicate section of the biopsy shown in A). Original
magnification: x400 in C; x1000 in D. (E and F)
Immunohistochemical analysis of CD3-positive T cells, illustrating the
distribution of peritubular T cells, which are characterized by homogeneous
dark staining of the cell bodies. Note the infiltration of the tubular
epithelium (arrowhead). Original magnification: x400 in E; x1000
in F.
CCR2B mRNA-expressing infiltrating mononuclear leukocytes werecommon in
the tubulointerstitium (Figure 7, C and
D). Thesecells were observed to be clustered around MCP-1
mRNA-positivetubular epithelium, as demonstrated by in situ
hybridizationof replicate tissue sections
(Figure 8, C and D).
Furthermore,CCR2B-positive cells were commonly arranged around crescentic
glomeruli(Figure 8, A and B).
There were significant correlations betweenthe mean numbers and localization
of interstitial CCR2B-positivecells and interstitial CD3-positive T cells
(Figures 7, C to F,and
9). Areas with interstitial
infiltrates organized aslymphoid follicles contained a few CCR2B-positive
cells, whichwere located mainly in the periphery of the follicles. Few CCR2B
mRNA-positivecells were observed within tubular lumina.
Figure 8. CCR2B mRNA expression in interstitial infiltrating mononuclear cells. (A
and B) In situ hybridization using a CCR2B antisense riboprobe,
demonstrating CCR2B mRNA expression in interstitial infiltrating cells, with
prominent clustering around the glomerulus. Original magnification: x
400 in A; x 1000 in B. (C and D) In situ hybridization using
antisense riboprobes for CCR2B (C) and MCP-1 (D) on replicate sections of a
crescentic glomerulonephritis biopsy. Original magnification: x 1000 in
C; x 1000 in D. Note the clustering of CCR2 mRNA-positive cells around a
tubule, with MCP-1 mRNA expression in the epithelium.
Figure 9. Correlation between the mean number of interstitial CD3-positive T cells
and the mean number of interstitial CCR2B mRNA-positive cells (P <
0.05).
CCR2B mRNA-expressing cells commonly infiltrated glomerulartufts and
crescents, but their numbers were lower than the numbersof glomerular
macrophages (Figure 10;
Table 2). Seven percentof the
total CCR2B mRNA-expressing cells were observed in glomeruli.Biopsies with
large numbers of CD68-positive macrophages inglomeruli and crescents
exhibited a trend toward larger numbersof glomerular CCR2B-expressing cells,
but the correlation didnot reach statistical significance
(r2 = 0.5, NS). No convincingCCR2B mRNA hybridization
signal was observed in intrinsic renalcells, but focal expression by
endothelial cells of small capillariescould not be excluded in areas with
extensive interstitial CCR2-positiveinfiltrates.
Figure 10. Glomerular MCP-1 and CCR2B mRNA expression. (A and C) In situ
hybridization using a CCR2B antisense riboprobe, demonstrating CCR2B mRNA
expression in infiltrating cells of a severely damaged glomerulus with strong
mononuclear cell infiltration. Original magnification: x 400 in A;
x 1000 in C. (B and D) In situ hybridization using a MCP-1
antisense riboprobe, demonstrating MCP-1 mRNA in a replicate section of the
biopsy shown in A. Original magnification: x 400 in B; x 1000 in
D. (E) Immunohistochemical analysis of CD68-positive macrophages in a
replicate section of the biopsy shown in A, demonstrating prominent glomerular
macrophage infiltration. Original magnification, x 400. (F)
Immunohistochemical analysis of CCR5-positive cells in a replicate section of
the biopsy shown in A. Note the weak diffuse CCR5 signal, with a distribution
similar to that of CD68-positive macrophages in E. Original magnification,
x 400.
CCR5-Positive Cells Are a Prominent Part of the Tubulointerstitial
Infiltrates, and the Mean Number Is Correlated with the Number of Interstitial
T Cells and Interstitial Macrophages
CCR5 expression was a prominent feature of interstitial infiltrating
leukocytes(Figure 1C). These
cells were commonly observed to be infiltratingthe tubular epithelium.
Clustering around Bowman's capsulesof inflamed glomeruli was also a common
feature for CCR5-positiveleukocytes. The best morphologic concordance was
observed betweenthe distributions of CD3-positive T cells and CCR5-positive
cells,which led to a significant correlation between the mean numberof
CCR5-positive cells and the mean number of CD3-positive Tcells (P
< 0.05). Furthermore, the numerical correlationbetween CCR5-positive cells
and interstitial CD68-positive macrophagesreached statistical significance
(P < 0.05). A morphologiccorrelation between areas of macrophage
infiltration and CCR5-positivecells was rarely observed
(Figure 11, G and H),
indicating asmall subset of CCR5-positive macrophages. CCR5-positive cells
withintubular lumina were rare, indicating that the intratubular
CD68-positivecells were mainly CCR5-negative.
Figure 11. Distribution of interstitial macrophages, T cells, and CCR2B- and
CCR5-positive cells. (A and E) Immunohistochemical analysis of CD3-positive T
cells, illustrating the distribution of interstitial T cells. Original
magnification: x 400 in A; x 1000 in E. (B and F) In situ
hybridization using a CCR2B antisense riboprobe, demonstrating CCR2B mRNA
expression in infiltrating cells in a serial section of the biopsy shown in A.
Original magnification: x 400 in B; x 1000 in F. (C and G)
Immunohistochemical analysis of CD68-positive macrophages, illustrating a
periglomerular layer surrounding a cellular crescent. Original magnification:
x 400 in C; x 1000 in G. (D and H) Immunohistochemical analysis of
CCR5 in a serial section from the biopsy shown in A. Original magnification:
x 400 in D; x 1000 in H. Note that the cell form and distribution
of CCR5-positive cells on the left side are better correlated with the T cell
distribution (compare A and D), whereas the lower section mirrors the
macrophage distribution (compare G and H).
Within glomeruli, CCR5-positive cells were rare but were detectablein a
subset of crescents (1.9% of the CCR5-positive cells).The CCR5 staining of
cells in crescents was generally weakerthan the staining of most of the
interstitial, CCR5-positive,infiltrating cells. The glomerular CD68-positive
cells outnumberedthe CCR5-positive cells. A glomerulus with an
extraordinarilylarge number of infiltrating cells and CCR5-positive cells
(themost encountered in this series) is illustrated in
Figure 10F.No CCR5 expression
was detectable in intrinsic renal cells.
The chemokine MCP-1 and its receptor CCR2 are likely involvedin the
recruitment of macrophages in inflammatory renal diseases.No data on the
distribution of CCR2-positive cells in humanrenal diseases are currently
available. cGN, which is the mostaggressive form of inflammatory glomerular
injury, is largelythe result of cell-mediated immune injury, including
prominentmacrophage infiltration of glomeruli
(2,7).
This is the firststudy to demonstrate the expression of CCR2B mRNA in
biopsiesfrom patients with cGN. CCR2B mRNA was expressed by infiltrating
mononuclearleukocytes, and CCR2B mRNA-expressing cells were common in
inflamedglomeruli and within crescents. The numbers and distributionof
glomerular CD68-positive monocytes/macrophages indicatedthat these cells were
the main glomerular cell type that expressedCCR2B mRNA. In addition, a
correlation between CCR2B mRNA-expressinginterstitial cells and similarly
located CD3-positive T cellsindicated that some CCR2B-positive T cells
infiltrated the interstitiumduring this disease process. In vitro
studies indicated thatmonocytes/macrophages
(22,23,24),
activated T cells
(25,26,27),
activatedB cells (28),
dendritic cells (29), and
activated natural killercells
(30,31)
express CCR2. CCR2 is a receptor for the chemokinesMCP-1 through -5
(18,32,33,34).
Two isoforms of CCR2 have beendescribed, i.e., CCR2A and CCR2B.
Currently, CCR2B is thoughtto be the major isoform expressed on cultured and
primary cells(35). The
expression of its ligand MCP-1 in renal diseases hasrecently been reviewed
(9). Our findings of MCP-1 mRNA
expressionby cells in glomerular tufts, crescents, and parietal epithelium,
aswell as by tubular epithelium and interstitial infiltratingleukocytes, are
in accord with previous studies
(36,37).
Largenumbers of glomerular MCP-1-expressing cells were associatedwith large
numbers of glomerular CD68-positive macrophages andCCR2B-positive cells, in
agreement with the concept of leukocytechemoattraction by MCP-1. In addition,
biopsies with large numbersof MCP-1 mRNA-positive cells contained
significantly largernumbers of CD3-positive T cells, at least some of which
exhibitedevidence of bearing the MCP-1 receptor CCR2B.
In our study, the morphologic distribution of MCP-1 expression,CCR2B
mRNA-positive cells, and CD68-positive macrophages isindicative of an
inflammatory process in which a subset of glomerularmacrophages are attracted
via CCR2B. Currently available dataon the role of CCR2 and MCP-1 in
nephrotoxic serum-induced nephritis,a model of human cGN, are not conclusive.
Several studies usingMCP-1-neutralizing antibodies have demonstrated
beneficial effectson glomerular lesions in rats
(38,39,40)
and mice (41). Studiesof
nephrotoxic serum-induced nephritis in rats demonstratedthat an approximately
35% decrease in glomerular macrophagescould be achieved by treatment with
anti-MCP-1 antibodies
(38,39,40).
Incontrast to this positive effect, which occurred 4 d after injury
initiation,Fujinaka et al.
(38) observed no differences
after 8 d. MCP-1-deficientmice exhibited no differences in glomerular lesions
in nephrotoxicserum-induced nephritis, compared with wild-type mice, but did
exhibita significant reduction in tubulointerstitial lesions
(42).CCR2-deficient mice
demonstrated a reduction in glomerular macrophageinfiltrates and proteinuria
after 24 h but exhibited a worsedisease course after 7 d
(43). A recent study of human
cGN demonstrateda correlation between the number of crescents and urinary
MCP-1excretion, as well as decreased urinary MCP-1 concentrationsafter
therapy (44).
In a mouse model, blockade of MCP-1 function resulted not onlyin a
reduction of the macrophage infiltration but also in asignificant reduction
of T cells (41). We observed a
significantcorrelation between the numbers of interstitial T cells and
CCR2B-positivecells, as well as significantly larger numbers of T cells,
whichwere localized in areas with more MCP-1-positive cells. Becausethe
ligands for CCR2 and CCR5 attract different subsets of Tcells, i.e.,
MCP-1 preferentially attracts CD4-positive T cellsand RANTES preferentially
attracts CD8-positive T cells
(45),the distribution of CCR2
and CCR5 mRNA (and presumably theirpeptide products) might reflect different
subpopulations ofCD3-positive cells in the renal interstitium in cGN.
We have demonstrated that CD3-positive T cells contribute tothe majority
of infiltrating cells in the interstitium but arerare in the injured
glomerular tufts in cGN and contribute littleto the cellular composition of
crescents. In contrast, macrophagesform a major part of the cells in
crescents, are often observedto be infiltrating the glomerular tuft, and are
also presentin large numbers in the interstitium in cGN. These data clearly
indicatedifferences in the infiltrating leukocytic cell populationsand in
the determinants of these different patterns of anatomiclocalization. The
mismatch between the number of T cells infiltratingglomeruli and the
interstitium has been well described
(5,13,15,46).
Inagreement with our data, Hooke et al.
(13,46)
observed no significantincrease in intraglomerular T cells in patients with
cGN. Weobserved a subset of crescents that contained three or moreT cells.
These were usually associated with loss of the capsuleseparating the
glomerulus and the interstitium, suggesting Tcell infiltration via the
interstitium rather than across glomerularcapillary walls. In contrast, the
distribution of monocytes/macrophages,which are commonly observed in inflamed
glomerular tufts andcrescents, indicates a greater tendency of these cells to
infiltrateand localize in glomeruli, compared with T cells.
The distribution of subsets of infiltrating leukocytes is reflectedin the
pattern of CCR2B- and CCR5-positive cells. As describedabove, CCR2B
mRNA-positive cells exhibited a high relative percentagewithin glomeruli. The
patterns of distribution and the overallpercentages of infiltrating
glomerular leukocytes indicate thatmonocytes/macrophages are the main
CCR2B-positive cell type.In contrast, the relative numbers of CCR5-positive
cells mirroredthe distribution of CD3-positive T cells, with small numbersin
the tuft and large numbers in the interstitium. In a previousstudy, we
demonstrated a similar distribution of T cells andCCR5-positive interstitial
cells in various noncrescentic glomerulardiseases
(15). In addition, a part of
the CCR5-positive interstitialcell population matched CD68-positive
macrophages with respectto both distribution and numbers. The number of
CCR5-positivecells within glomeruli is very small, and the number of
glomerularmacrophages is far greater than the number of CCR5-positivecells.
This suggests either that there is a population of CCR5-negativemacrophages
that differentially infiltrate glomeruli, ratherthan the tubulointerstitium,
or that CCR5 on glomerular macrophagesis downregulated, rendering it
undetectable by our methods.
This is the first study of the distribution of CCR2B mRNA-expressingcells
in the context of MCP-1 expression, CCR5-positive cells,T cells, and
macrophages in human cGN. We propose that the differentialexpression of
chemokines and their receptors mediates, at leastin part, different patterns
of leukocyte influx into renal parenchymalcompartments in acute injury, such
as cGN. CCR2B and its ligandsseem to be important for glomerular macrophage
infiltration,as well as for attraction of a subset of interstitial T cells.
Incontrast, engagement of CCR5 seems to promote tubulointerstitial
infiltrationby T cells and a smaller subset of macrophages. These data
indicatethat therapeutic approaches that block either CCR2 or CCR5 could
interveneat different steps in the injury process.
Acknowledgments
This work was supported by a grant to the O'Brien Kidney ResearchCenter
(National Institutes of Health Grant DK47659), GrantHL63652 from the National
Institutes of Health, and a grantfrom the Else
Kröner-Fresenius-Stiftung (Bad Homburg v.d.
Höhe, Germany).
Alpers CE, Hudkins KL, Floege J, Johnson RJ: Human renal cortical
interstitial cells with some features of smooth muscle cells participate in
tubulointerstitial and crescentic glomerular injury. J Am Soc
Nephrol 5:201
-209, 1994[Abstract]
Ferrario F, Rastaldi MP: Necrotizing-crescentic glomerulonephritis
in ANCA-associated vasculitis: The role of monocytes. Nephrol Dial
Transplant 14:1627
-1631, 1999[Free Full Text]
Hudkins KL, Giachelli CM, Eitner F, Couser WG, Johnson RJ, Alpers
CE: Osteopontin expression in human crescentic glomerulonephritis.
Kidney Int 57:105
-116, 2000[Medline]
Cunningham MA, Huang XR, Dowling JP, Tipping PG, Holdsworth SR:
Prominence of cell-mediated immunity effectors in "pauciimmune"
glomerulonephritis. J Am Soc Nephrol10
: 499-506,1999[Abstract/Free Full Text]
Springer TA: Traffic signals for lymphocyte recirculation and
leukocyte emigration: The multistep paradigm. Cell76
: 301-314,1994[Medline]
Segerer S, Nelson PJ, Schlondorff D: Chemokines, chemokine
receptors, and renal disease: From basic science to pathophysiologic and
therapeutic studies. J Am Soc Nephrol11
: 152-176,2000[Abstract/Free Full Text]
Luster AD: Chemokines: Chemotactic cytokines that mediate
inflammation. N Engl J Med 338:436
-445, 1998[Free Full Text]
Wenzel UO, Abboud HE: Chemokines and renal disease. Am J
Kidney Dis 26:982
-994, 1995[Medline]
Hooke DH, Gee DC, Atkins RC: Leukocyte analysis using monoclonal
antibodies in human glomerulonephritis. Kidney Int31
: 964-972,1987[Medline]
Bohle A, Wehrmann M, Bogenschutz O, Batz C, Vogl W, Schmitt H,
Muller CA, Muller GA: The long-term prognosis of the primary
glomerulonephritides: A morphological and clinical analysis of 1747 cases.
Pathol Res Pract 188:908
-924, 1992[Medline]
Segerer S, Mack M, Regele H, Kerjaschki D, Schlondorff D:
Expression of the C-C chemokine receptor 5 in human kidney diseases.
Kidney Int 56:52
-64, 1999[Medline]
Deng H, Liu R, Ellmeier W, Choe S, Unutmaz D, Burkhart M, Di Marzio
P, Marmon S, Sutton RE, Hill CM, Davis CB, Peiper SC, Schall TJ, Littman DR,
Landau NR: Identification of a major co-receptor for primary isolates of
HIV-1. Nature (Lond.) 381:661
-666, 1996[Medline]
Isik FF, Rand RP, Gruss JS, Benjamin D, Alpers CE: Monocyte
chemoattractant protein-1 mRNA expression in hemangiomas and vascular
malformations. J Surg Res 61:71
-76, 1996[Medline]
Charo IF, Myers SJ, Herman A, Franci C, Connolly AJ, Coughlin SR:
Molecular cloning and functional expression of two monocyte chemoattractant
protein 1 receptors reveals alternative splicing of the carboxyl-terminal
tails. Proc Natl Acad Sci USA91
: 2752-2756,1994[Abstract/Free Full Text]
Eitner F, Cui Y, Hudkins KL, Alpers CE: Chemokine receptor (CXCR4)
mRNA-expressing leukocytes are increased in human renal allograft rejection.
Transplantation 66:1551
-1557, 1998[Medline]
Eitner F, Cui Y, Hudkins KL, Anderson DM, Schmidt A, Morton WR,
Alpers CE: Chemokine receptor (CCR5) expression in human kidneys and in the
HIV infected macaque. Kidney Int54
: 1945-1954,1998[Medline]
Alpers CE, Hudkins KL, Davis CL, Marsh CL, Riches W, McCarty JM,
Benjamin CD, Carlos TM, Harlan JM, Lobb R: Expression of vascular cell
adhesion molecule-1 in kidney allograft rejection. Kidney
Int 44: 805-816,1993[Medline]
Penton-Rol G, Polentarutti N, Luini W, Borsatti A, Mancinelli R,
Sica A, Sozzani S, Mantovani A: Selective inhibition of expression of the
chemokine receptor CCR2 in human monocytes by IFN-. J
Immunol 160:3869
-3873, 1998[Abstract/Free Full Text]
Penton-Rol G, Cota M, Polentarutti N, Luini W, Bernasconi S,
Borsatti A, Sica A, LaRosa GJ, Sozzani S, Poli G, Mantovani A: Up-regulation
of CCR2 chemokine receptor expression and increased susceptibility to the
multitropic HIV strain 89.6 in monocytes exposed to glucocorticoid hormones.
J Immunol 163:3524
-3529, 1999[Abstract/Free Full Text]
Fantuzzi L, Borghi P, Ciolli V, Pavlakis G, Belardelli F, Gessani
S: Loss of CCR2 expression and functional response to monocyte chemotactic
protein (MCP-1) during the differentiation of human monocytes: Role of
secreted MCP-1 in the regulation of the chemotactic response.
Blood 94:875
-883, 1999[Abstract/Free Full Text]
Ruffing N, Sullivan N, Sharmeen L, Sodroski J, Wu L: CCR5 has an
expanded ligand-binding repertoire and is the primary receptor used by MCP-2
on activated T cells. Cell Immunol189
: 160-168,1998[Medline]
Loetscher P, Seitz M, Baggiolini M, Moser B: Interleukin-2
regulates CC chemokine receptor expression and chemotactic responsiveness in T
lymphocytes. J Exp Med 184:569
-577, 1996[Abstract/Free Full Text]
Sallusto F, Kremmer E, Palermo B, Hoy A, Ponath P, Qin S, Forster
R, Lipp M, Lanzavecchia A: Switch in chemokine receptor expression upon TCR
stimulation reveals novel homing potential for recently activated T cells.
Eur J Immunol 29:2037
-2045, 1999[Medline]
Kim CH, Broxmeyer HE: Chemokines: Signal lamps for trafficking of T
and B cells for development and effector function. J Leukocyte
Biol 65: 6-15,1999[Abstract]
Polentarutti N, Allavena P, Bianchi G, Giardina G, Basile A,
Sozzani S, Mantovani A, Introna M: IL-2-regulated expression of the monocyte
chemotactic protein-1 receptor (CCR2) in human NK cells: Characterization of a
predominant 3.4-kilobase transcript containing CCR2B and CCR2A sequences.
J Immunol 158:2689
-2694, 1997[Abstract]
Sozzani S, Introna M, Bernasconi S, Polentarutti N, Cinque P, Poli
G, Sica A, Mantovani A: MCP-1 and CCR2 in HIV infection: Regulation of agonist
and receptor expression. J Leukocyte Biol62
: 30-33,1997[Abstract]
Franci C, Wong LM, Van Damme J, Proost P, Charo IF: Monocyte
chemoattractant protein-3, but not monocyte chemoattractant protein-2, is a
functional ligand of the human monocyte chemoattractant protein-1 receptor.
J Immunol 154:6511
-6517, 1995[Abstract]
Berkhout TA, Sarau HM, Moores K, White JR, Elshourbagy N, Appelbaum
E, Reape RJ, Brawner M, Makwana J, Foley JJ, Schmidt DB, Imburgia C, McNulty
D, Matthews J, O'Donnell K, O'Shannessy D, Scott M, Groot PHE, Macphee C:
Cloning, in vitro expression, and functional characterization of a
novel human CC chemokine of the monocyte chemotactic protein (MCP) family
(MCP-4) that binds and signals through the CC chemokine receptor 2B.
J Biol Chem 272:16404
-16413, 1997[Abstract/Free Full Text]
Gong X, Gong W, Kuhns DB, Ben-Baruch A, Howard OM, Wang JM:
Monocyte chemotactic protein-2 (MCP-2) uses CCR1 and CCR2B as its functional
receptors. J Biol Chem 272:11682
-11685, 1997[Abstract/Free Full Text]
Charo IF: CCR2: From cloning to the creation of knockout mice.
Chem Immunol 72:30
-41, 1999[Medline]
Rovin BH, Rumancik M, Tan L, Dickerson J: Glomerular expression of
monocyte chemoattractant protein-1 in experimental and human
glomerulonephritis. Lab Invest71
: 536-542,1994[Medline]
Cockwell P, Howie AJ, Adu D, Savage CO: In situ analysis
of C-C chemokine mRNA in human glomerulonephritis. Kidney
Int 54: 827-836,1998[Medline]
Fujinaka H, Yamamoto T, Takeya M, Feng L, Kawasaki K, Yaoita E,
Kondo D, Wilson CB, Uchiyama M, Kihara I: Suppression of anti-glomerular
basement membrane nephritis by administration of anti-monocyte chemoattractant
protein-1 antibody in WKY rats. J Am Soc Nephrol8
: 1174-1178,1997[Abstract]
Tang WW, Qi M, Warren JS: Monocyte chemoattractant protein 1
mediates glomerular macrophage infiltration in anti-GBM Ab GN.
Kidney Int 50:665
-671, 1996[Medline]
Wada T, Yokoyama H, Furuichi K, Kobayashi KI, Harada K, Naruto M,
Su SB, Akiyama M, Mukaida N, Matsushima K: Intervention of crescentic
glomerulonephritis by antibodies to monocyte chemotactic and activating factor
(MCAF/MCP-1). FASEB J 10:1418
-1425, 1996[Abstract]
Lloyd CM, Minto AW, Dorf ME, Proudfoot A, Wells TN, Salant DJ,
Gutierrez-Ramos JC: RANTES and monocyte chemoattractant protein-1 (MCP-1) play
an important role in the inflammatory phase of crescentic nephritis, but only
MCP-1 is involved in crescent formation and interstitial fibrosis.
J Exp Med 185:1371
-1380, 1997[Abstract/Free Full Text]
Tesch GH, Schwarting A, Kinoshita K, Lan HY, Rollins BJ, Kelley VR:
Monocyte chemoattractant protein-1 promotes macrophage-mediated tubular
injury, but not glomerular injury, in nephrotoxic serum nephritis.
J Clin Invest 103:73
-80, 1999[Medline]
Bird JE, Giancarli MR, Kurihara T, Kowala MC, Valentine MT, Gitlitz
PH, Pandya DG, French MH, Durham SK: Increased severity of glomerulonephritis
in C-C chemokine receptor 2 knockout mice. Kidney Int57
: 129-136,2000[Medline]
Wada T, Furuichi K, Segawa-Takaeda C, Shimizu M, Sakai N, Takeda
SI, Takasawa K, Kida H, Kobayashi KI, Mukaida N, Ohmoto Y, Matsushima K,
Yokoyama H: MIP-1 and MCP-1 contribute to crescents and interstitial
lesions in human crescentic glomerulonephritis. Kidney
Int 56: 995-1003,1999[Medline]
Rabin RL, Park MK, Liao F, Swofford R, Stephany D, Farber JM:
Chemokine receptor responses on T cells are achieved through regulation of
both receptor expression and signaling. J Immunol162
: 3840-3850,1999[Abstract/Free Full Text]
Hooke DH, Hancock WW, Gee DC, Kraft N, Atkins RC: Monoclonal
antibody analysis of glomerular hypercellularity in human glomerulonephritis.
Clin Nephrol 22:163
-168, 1984[Medline]
Received for publication February 29, 2000.
Accepted for publication May 17, 2000.
This article has been cited by other articles:
J. Menke, G. C. Zeller, E. Kikawada, T. K. Means, X. R. Huang, H. Y. Lan, B. Lu, J. Farber, A. D. Luster, and V. R. Kelley CXCL9, but not CXCL10, Promotes CXCR3-Dependent Immune-Mediated Kidney Disease
J. Am. Soc. Nephrol.,
June 1, 2008;
19(6):
1177 - 1189.
[Full Text][PDF]
S. Segerer, R. Djafarzadeh, H.-J. Grone, C. Weingart, D. Kerjaschki, C. Weber, A. J. Kungl, H. Regele, A. E.I. Proudfoot, and P. J. Nelson Selective Binding and Presentation of CCL5 by Discrete Tissue Microenvironments during Renal Inflammation
J. Am. Soc. Nephrol.,
June 1, 2007;
18(6):
1835 - 1844.
[Abstract][Full Text][PDF]
S. Campbell, L. C. Burkly, H.-X. Gao, J. W. Berman, L. Su, B. Browning, T. Zheng, L. Schiffer, J. S. Michaelson, and C. Putterman Proinflammatory Effects of Tweak/Fn14 Interactions in Glomerular Mesangial Cells
J. Immunol.,
February 1, 2006;
176(3):
1889 - 1898.
[Abstract][Full Text][PDF]
U. Panzer, O. M. Steinmetz, R. R. Reinking, T. N. Meyer, S. Fehr, A. Schneider, G. Zahner, G. Wolf, U. Helmchen, P. Schaerli, et al. Compartment-Specific Expression and Function of the Chemokine IP-10/CXCL10 in a Model of Renal Endothelial Microvascular Injury
J. Am. Soc. Nephrol.,
February 1, 2006;
17(2):
454 - 464.
[Abstract][Full Text][PDF]
R. Birck, M. Newman, C. Braun, I. Neumann, K. Nemoto, B. Yard, R. Waldherr, and F. J. van der Woude 15-Deoxyspergualin and cyclophosphamide, but not mycophenolate mofetil, prolong survival and attenuate renal disease in a murine model of ANCA-associated crescentic nephritis
Nephrol. Dial. Transplant.,
January 1, 2006;
21(1):
58 - 63.
[Abstract][Full Text][PDF]
K. Kitagawa, T. Wada, K. Furuichi, H. Hashimoto, Y. Ishiwata, M. Asano, M. Takeya, W. A. Kuziel, K. Matsushima, N. Mukaida, et al. Blockade of CCR2 Ameliorates Progressive Fibrosis in Kidney
Am. J. Pathol.,
July 1, 2004;
165(1):
237 - 246.
[Abstract][Full Text][PDF]
T. Wada, K. Furuichi, N. Sakai, Y. Iwata, K. Kitagawa, Y. Ishida, T. Kondo, H. Hashimoto, Y. Ishiwata, N. Mukaida, et al. Gene Therapy via Blockade of Monocyte Chemoattractant Protein-1 for Renal Fibrosis
J. Am. Soc. Nephrol.,
April 1, 2004;
15(4):
940 - 948.
[Abstract][Full Text][PDF]
K. Furuichi, T. Wada, Y. Iwata, K. Kitagawa, K.-i. Kobayashi, H. Hashimoto, Y. Ishiwata, M. Asano, H. Wang, K. Matsushima, et al. CCR2 Signaling Contributes to Ischemia-Reperfusion Injury in Kidney
J. Am. Soc. Nephrol.,
October 1, 2003;
14(10):
2503 - 2515.
[Abstract][Full Text][PDF]
Z.-H. Liu, S.-F. Chen, H. Zhou, H.-P. Chen, and L.-S. Li Glomerular expression of C-C chemokines in different types of human crescentic glomerulonephritis
Nephrol. Dial. Transplant.,
August 1, 2003;
18(8):
1526 - 1534.
[Abstract][Full Text][PDF]
Z.-H. Liu, S.-F. Chen, H. Zhou, H.-P. Chen, and L.-S. Li Glomerular expression of C-C chemokines in different types of human crescentic glomerulonephritis
Nephrol. Dial. Transplant.,
August 1, 2003;
18(88):
1526 - 1534.
[Abstract][Full Text]
C. Viedt and S. R. Orth Monocyte chemoattractant protein-1 (MCP-1) in the kidney: does it more than simply attract monocytes?
Nephrol. Dial. Transplant.,
December 1, 2002;
17(12):
2043 - 2047.
[Full Text][PDF]
G. Wolf, T. Jocks, G. Zahner, U. Panzer, and R. A. K. Stahl Existence of a regulatory loop between MCP-1 and TGF-beta in glomerular immune injury
Am J Physiol Renal Physiol,
November 1, 2002;
283(5):
F1075 - F1084.
[Abstract][Full Text][PDF]
S. Segerer, F. Eitner, Y. Cui, K. L. Hudkins, and C. E. Alpers Cellular Injury Associated with Renal Thrombotic Microangiopathy in Human Immunodeficiency Virus-Infected Macaques
J. Am. Soc. Nephrol.,
February 1, 2002;
13(2):
370 - 378.
[Abstract][Full Text][PDF]
S. Taneda, S. Segerer, K. L. Hudkins, Y. Cui, M. Wen, M. Segerer, M. H. Wener, C. G. Khairallah, A. G. Farr, and C. E. Alpers Cryoglobulinemic Glomerulonephritis in Thymic Stromal Lymphopoietin Transgenic Mice
Am. J. Pathol.,
December 1, 2001;
159(6):
2355 - 2369.
[Abstract][Full Text][PDF]
H.-J. ANDERS, V. VIELHAUER, M. KRETZLER, C. D. COHEN, S. SEGERER, B. LUCKOW, L. WELLER, H.-J. GRÖNE, and D. SCHLÖNDORFF Chemokine and Chemokine Receptor Expression during Initiation and Resolution of Immune Complex Glomerulonephritis
J. Am. Soc. Nephrol.,
May 1, 2001;
12(5):
919 - 931.
[Abstract][Full Text]
Top
Abstract
Introduction
. J
Immunol 160:3869
-3873, 1998
and MCP-1 contribute to crescents and interstitial
lesions in human crescentic glomerulonephritis. Kidney
Int 56: 995-1003,1999[Medline]
