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Production by Intrinsic Renal Cells and Bone Marrow-Derived Cells Is Required for Full Expression of Crescentic Glomerulonephritis in Mice1
Center for Inflammatory Diseases, Monash University, and Department of Medicine, Monash Medical Center, Clayton, Victoria, Australia
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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from bone marrow (BM) and
non-BM-derived cells to glomerular and cutaneous delayed-type
hypersensitivity (DTH) was studied in mice. Chimeric IFN- mice
(IFN-+/+ BM chimera), in which IFN- production was
restricted to BM-derived cells, were created by transplanting normal
C57BL/6 (wild-type (WT)) BM into irradiated IFN--deficient mice. BM
IFN--deficient chimeric mice (IFN--/- BM chimera)
were created by transplanting WT mice with IFN--deficient BM. WT and
sham chimeric mice (WT mice transplanted with WT BM) developed
crescentic glomerulonephritis (GN) with features of DTH (including
glomerular T cell and macrophage infiltration) in response to an Ag
planted in their glomeruli and skin DTH following subdermal Ag
challenge. IFN--deficient mice showed significant protection from
crescentic GN and reduced cutaneous DTH. IFN-+/+ BM
chimeric and IFN--/- BM chimeric mice showed similar
attenuation of crescentic GN as IFN--deficient mice, whereas
cutaneous DTH was reduced only in IFN--/- BM chimeras.
In crescentic GN, IFN- was expressed by tubular cells and occasional
glomerular cells and was colocalized with infiltrating CD8+
T cells, but not with CD4+ T cells or macrophages. Renal
MHC class II expression was reduced in IFN-+/+ BM
chimeric mice and was more severely reduced in IFN--deficient mice
and IFN--/- BM chimeric mice. These studies show that
IFN- expression by both BM-derived cells and intrinsic renal cells
is required for the development of crescentic GN, but IFN-
production by resident cells is not essential for the development of
cutaneous DTH. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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) or
administration of Th2 cytokines (IL-4 and IL-10) attenuates crescentic
anti-glomerular basement membrane (GBM)-induced GN in mice by
attenuating the systemic Th1 immune responses to the nephritogenic Ag.
The contribution of cytokines produced by intrinsic renal cells to the
effector phase of immune renal injury is poorly defined. We have
recently demonstrated that local IL-12 production by intrinsic renal
cells as well as production by bone marrow (BM)-derived cells
contribute to inflammatory renal injury in this model
(2).
IFN- is a potent proinflammatory molecule that activates macrophages
and up-regulates MHC class I and MHC class II (MHC II) expression. It
augments Th1 responses by inducing IL-12R expression on T cells
(3) and enhancing IL-12 production by macrophages
(4). IFN- is produced predominately by
CD8+ T cells, CD4+ Th1
cells, and NK cells (5), but a variety of other cells,
including endothelial cells (6, 7, 8) and smooth muscle cells
(9), have also been demonstrated to be capable of
producing this cytokine. IFN--deficient mice produced by targeted
mutation of the IFN- gene show normal development of the immune
system and are healthy in the absence of infection (10).
However, they are highly susceptible to intracellular pathogens and
show markedly reduced MHC II expression, NO and superoxide production
by macrophages (10), and reduced Th1 Ab isotypes and
cutaneous DTH following Ag challenge (11, 12, 13).
IFN- is expressed in glomeruli in human (14, 15) and
experimental GN (16, 17). The functional contribution of
IFN- to the development of glomerular injury has been demonstrated
by attenuation of crescentic GN by in vivo Ab inhibition of IFN-
(16) and by using IFN--deficient mice
(11). IFN- receptor-deficient mice also show
attenuation of renal injury induced by anti-GBM Ab
(17). An important mechanism for the protection afforded
by blocking or the absence of IFN- is via attenuation of the
systemic Th1-biased immune response to the nephritogenic Ag, manifest
by changes in the Ab isotype profile and diminution of cutaneous DTH
response (11). This effect was not observed in a
noncrescentic model of anti-GBM GN associated with mild glomerular
injury (18). In this model IFN--deficient mice were
more prone to injury (31%) than wild-type (WT) mice (7%), but IFN-
deficiency did not alter the Th1 bias of the immune response to the
nephritogenic Ag. In addition to its ability to direct the systemic Th
subset toward Th1, IFN- has the potential to augment GN by local
effects on intrinsic renal cells. IFN- stimulates MHC class I
(19) and II (20), IL-1 (21), and
monocyte chemoattractant protein-1 (22) expression by
mesangial cells in vitro. It induces MHC II expression on tubular
epithelial cells (23, 24, 25, 26, 27) and induces MHC class I and II
and ICAM-1 on glomerular epithelial cells (28, 29).
Although IFN- is expressed in the human kidney, and experimental GN
and intrinsic renal cells respond to IFN- in vitro, their capacity
to produce IFN- has not been demonstrated. The demonstration of
IFN- production by endothelial cells (6, 7, 8) and smooth
muscle cells (9) that are from a similar lineage as some
renal cells (renal endothelial cells and mesangial cells) suggests the
potential for IFN- production by intrinsic renal cells during
inflammation. The current study demonstrates this capacity and examines
the contribution of IFN- from BM-derived and non-BM-derived cells to
the development of DTH responses in the kidney and skin. It
demonstrates a role for both sources of IFN- in the development of
Th1-dependent glomerular DTH, but only for BM cell-derived IFN- in
cutaneous DTH.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Breeding pairs of mice with a targeted disruption of the IFN-
gene (10) were obtained from The Jackson Laboratory
(Bar Harbor, ME). These mice have been back-crossed for nine
generations onto a C57BL/6 background. Mice were housed and bred under
specific pathogen-free conditions at Monash University (Clayton,
Australia).
BM transplantation
Five- to 6-wk-old male homozygous IFN--deficient
(IFN--/-) or C57BL/6 (WT) recipient mice
received 1100-rad total body irradiation. BM cells were harvested
aseptically from the femurs and tibiae of WT or
IFN--/- donor mice and depleted of RBCs.
Recipient mice were injected i.v. with 5 x
106 cells within 6 h of irradiation. Mice
were housed under specific pathogen-free conditions for 8 wk to allow
BM reconstitution. Circulating leukocyte numbers and lymphocyte subsets
were assessed by flow cytometry as previously described
(2) and were normal 8 wk after BM transplantation. The
splenic distribution of T cells and macrophages, assessed by
immunohistology, also appeared normal at this time, as previously
reported (2). The efficiency of BM replacement following
transplantation was assessed using congenic CD45 mouse strains (Ly5.1
and Ly5.2). Eight weeks after irradiation and transplantation 92
± 1% of circulating leukocytes, 98 ± 1% of BM leukocytes, and
90 ± 2% of splenic leukocytes expressed the phenotypic marker of
the transplanted BM (n = 6), similar to previous
reports (30, 31). Resident CD45-positive cells of either
strain were rarely detectable by immunofluorescence in kidneys of
normal mice or mice that had undergone BM transplantation before
induction of GN.
Induction of crescentic GN
Crescentic anti-GBM was induced in male mice by i.v.
injection of a total of 14.4 mg sheep anti-mouse GBM globulin (in
900 µl PBS) divided equally into two doses given 3 h apart. The
development of crescentic GN and cutaneous DTH was assessed in WT and
IFN--/- mice and the three groups of
irradiated BM transplanted chimeric mice. These included sham chimeras
in which WT BM was transplanted into WT mice,
IFN-+/+ BM chimeras in which WT BM was
transplanted into IFN--deficient mice, and
IFN--/- BM chimeras in which
IFN--/- BM was transplanted into WT mice.
All mice were 13–14 wk of age at the time of administration of
anti-GBM globulin. Renal injury and cutaneous DTH were assessed 21
days later.
Histological assessment of glomerular injury
Glomerular crescent formation. Kidney tissue was fixed in Bouin’s fixative and embedded in paraffin, and 3-µm sections were stained with periodic acid-Schiff reagent. Glomeruli were considered to exhibit crescent formation when two or more layers of cells were observed in Bowman’s space. A minimum of 50 glomeruli were assessed to determine the crescent score for each animal.
Glomerular CD4+ cell, CD8+ cell, B cell, NK cell, and macrophage accumulation. Kidney tissue and spleen were fixed in periodate/lysine/paraformaldehyde. Six-micrometer cryostat-cut sections were stained to demonstrate CD4+ cells, CD8+ cells, B cells, and macrophages with mAbs, GK1.5, 53-6.7, 145-2C11, and M/170, respectively, using a three-layer immunoperoxidase technique (32). NK cells were detected with a polyclonal anti-asialo GM Ab (Bio-Scientific, Gymea, Australia) followed by a three-layer immunoperoxidase technique. A minimum of 20 equatorially sectioned glomeruli were assessed per animal, and results were expressed as cells per glomerular cross-section (c/gcs).
Tubulointerstitial infiltration. The number of interstitial cells in periodic acid-Schiff-stained renal tissue sections was counted using a 10-mm2 graticule fitted in the eyepiece of the microscope. Five randomly selected cortical areas, which excluded glomeruli, were counted for each animal. Each high-powered field represented an area of 1 mm2. Data are expressed as cells per square millimeter and are the mean ± SEM for four animals in each group.
Functional assessment of glomerular injury
Proteinuria. Mice were housed individually in cages to collect urine before administration of anti-GBM globulin and over the final 24 h of the experiment. Urinary protein concentrations were determined by a modified Bradford method. Before induction of GN, all groups of mice had 24-h urinary protein excretion in the normal range (0.5–2 mg/24 h).
Serum creatinine. Serum creatinine concentrations were measured by the alkaline picric acid method using an autoanalyzer (Roche Diagnostic Systems, Melbourne, Australia).
Demonstration of renal and splenic MHC II and IFN- expression by
confocal microscopy
Protein G-purified rat anti-mouse MHC II mAb (clone Y3P, a
gift from Prof. K. Shortman, Walter and Eliza Hall Institute,
Melbourne, Australia) was conjugated with Alexa Fluor dye 488
(Molecular Probes, Eugene, OR) as previously described
(2). Protein G-purified anti-mouse CD4 mAb (GK1.5) and
rat anti-mouse macrophage Ab (M170) were conjugated to Alexa Fluor
dye 594 (Molecular Probes). Allophycocyanin-conjugated rat
anti-mouse CD8a (Ly-2) was obtained from BD PharMingen (San Diego,
CA). FITC-conjugated rat anti-mouse IFN- (clone XMG1.2) was
obtained from Caltag Laboratories (Burlingame, CA). Cryostat-cut,
snap-frozen kidney tissue sections (6 µm) were blocked with 5%
normal rat serum in 1% BSA/PBS and then incubated with both Abs at a
final dilution of 1/50 for 60 min at room temperature. Confocal images
were collected using a Bio-Rad confocal inverted Nikon microscope
(Bio-Rad, Hercules, CA) equipped with an air-cooled 25-mW argon/krypton
laser, as previously described (2).
Induction of cutaneous DTH to sheep globulin
Mice developing GN were challenged 24 h before the end of
the experiment by intradermal injection of 500 µg sheep globulin in
50 µl PBS into one hind footpad. A similar dose of an irrelevant Ag
(horse globulin) was injected into the other footpad as a control.
Footpad swelling was quantified 24 h later using a micrometer
(Mitutoyo, Kawasaki-shi, Japan). Ag-specific DTH was taken as
the difference in skin swelling between sheep globulin- and horse
globulin-injected footpads and expressed as change in footpad
thickness (
millimeters).
Humoral immune responses to sheep globulin
Mouse anti-sheep globulin Ab titers were measured by ELISA on serum collected at the end of each experiment. Assays were performed using microtiter plates coated with sheep globulin at a concentration of 10 µg/ml, and bound mouse Ig was detected using HRP-conjugated sheep anti-mouse Ig (Amersham, Little Chalfont, U.K.) as the detecting Ab, as previously described (33).
Experimental design and statistical analysis
BM engraftment and lymphocyte subset reconstitution were
assessed on two separate occasions in a total of 16 sham chimeric mice,
13 IFN-+/+ BM chimeric mice, and 5
IFN--/- BM chimeric mice. Age-matched WT
mice served as controls. The development of GN was studied in a total
of 21 WT mice, 19 IFN--/- mice, 17 sham
chimeric mice, 13 IFN-+/+ BM chimeric mice,
and 11 IFN--/- BM chimeric mice. Results are
expressed as the mean ± SEM. Statistical significance was
determined by one-way ANOVA, followed by Newman-Keuls post-hoc
test.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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-deficient mice
WT mice developed proliferative GN with crescent formation (Fig. 1
). Sham chimeras developed disease of
similar severity, indicating that BM transplantation per se did not
affect the development of this disease. The incidences of crescentic
glomeruli (WT, 22.3 ± 1.4%; sham chimeras, 23.5 ± 2.6%),
glomerular infiltration of CD4+ cells (WT,
1.1 ± 0.1 c/gcs; sham chimeras, 1 ± 0.1 c/cgs), and
macrophages (WT, 2.4 ± 0.1 c/gcs; sham chimeras, 2.3 ± 0.3
c/gcs; Fig. 2) were equivalent in both groups.
CD8+ cells were also observed in glomeruli
(0.9 ± 0.1c/gcs) and in interstitial areas; however, NK cells
(0.05 ± 0.01 c/gcs) and B cells (0.01 ± 0.01 c/gcs) were
rarely observed. The interstitial cellular infiltrate (WT, 146 ±
6.2 cells/mm2; sham chimeras, 149 ± 7.5
cells/mm2), consisting mainly of macrophages with
occasional CD4+ and CD8+
cells, was also unaffected by BM transplantation. NK cells and B cells
were not observed in the interstitium. Similarly, there was no
difference in functional renal injury between the two groups, indicated
by proteinuria (WT, 9.6 ± 1.4 mg/24 h; sham chimeras, 7.8 ±
0.6 mg/24 h) and serum creatinine (WT, 23.9 ± 2.1 µmol/l; sham
chimeras, 17.4 ± 1.5 µmol/l; Fig. 3).
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-deficient mice. Glomerular crescent formation
(7.4 ± 0.8% of glomeruli) was attenuated compared with that in
WT mice (p < 0.001), as was glomerular
infiltration of CD4+ cells (0.5 ± 0.1
c/gcs; p < 0.001), macrophages (0.8 ± 0.2 c/gcs;
p < 0.001), and the interstitial cell infiltrate
(103 ± 6 cells/mm2; p <
0.001; Fig. 2
). Glomerular infiltration
of CD8+ cells (0.7 ± 0.3 c/gcs), NK cells
(0.04 ± 0.01 c/gcs), and B cells (0.04 ± 0.02 c/gcs) was
unaffected. A reduction in the histological severity of injury (Fig. 1)
was associated with significant protection of renal function, indicated
by reduction in proteinuria (3.3 ± 0.7 mg/24 h; p
< 0.01) and serum creatinine (14 ± 0.9 µmol/l;
p < 0.01) in IFN- mice compared with WT mice (Fig. 3).
IFN- production by intrinsic renal cells and BM-derived cells is
required for full expression of crescentic GN
Compared with WT mice and sham chimeras, the development of
crescentic GN was significantly attenuated in
IFN-+/+ BM chimeric and
IFN--/- BM chimeric mice, indicating that
IFN- from both intrinsic renal cells and BM-derived cells is
required for full expression of immune renal injury in this model. The
incidence of crescentic glomeruli was significantly reduced
(IFN-+/+ BM chimeras, 10.8 ± 1.2%
(p < 0.001);
IFN--/- BM chimeras, 12 ± 1%
(p < 0.001)). Glomerular accumulation of
CD4+ cells (IFN-+/+ BM
chimeras, 0.6 ± 0.1 c/gcs (p < 0.001);
IFN--/- BM chimeras, 0.1 ± 0.03 c/gcs
(p < 0.001)), CD8+ cells
(IFN-+/+ BM chimeras, 0.4 ± 0.06 c/gcs
(p < 0.05); IFN--/-
BM chimeras, 0.2 ± 0.03 c/gcs (p <
0.001)), and macrophages (IFN-+/+ BM chimeras,
1.1 ± 0.2 c/gcs (p < 0.01);
IFN--/- BM chimeras, 0.4 ± 0.1
(p < 0.001)) and the interstitial inflammatory
infiltrate (IFN-+/+ BM chimeras, 118 ± 5
cells/mm2 (p < 0.01);
IFN--/- BM chimeras, 67 ± 4
cells/mm2 (p < 0.001))
were all significantly reduced (Fig. 2). The sparse appearance of NK
cells and B cells was unaffected. Renal function was preserved in
chimeric mice with significantly less proteinuria
(IFN-+/+ BM chimeras, 5.3 ± 0.7 mg/24 h
(p < 0.01); IFN--/-
BM chimeras, 2.7 ± 0.4 mg/24 h (p <
0.01)) and lower serum creatinine levels
(IFN-+/+ BM chimeras, 14.6 ± 1.5
µmol/l (p < 0.01);
IFN--/- BM chimeras, 15.7 ± 1.4
µmol/l (p < 0.05)) compared with WT mice
with GN (Fig. 3).
Splenic and renal expression of IFN- (Fig. 4)
IFN- (Fig. 4, green) was strongly expressed in the
spleen and kidneys of WT mice with GN. In the spleen, coexpression
(Fig. 4A, yellow) of IFN- with CD4+
cells was observed on many cells, although some
CD4+ cells (Fig. 4A, red) did not
express IFN-. In the kidney (Fig. 4B), IFN- expression
was observed on glomerular cells, tubular cells, and interstitial
cells. CD4+ cells were prominent in glomeruli,
but coexpression of IFN- with CD4+ was not
observed. However, IFN- expression was colocalized (Fig. 4C, yellow) on CD8+ cells in
interstitial areas of WT mice. In IFN--deficient mice
CD4+ cells were observed in the spleen (Fig. 4D), but no staining for IFN- was observed, confirming
the specificity of the anti-IFN- Ab. In the kidneys of
IFN--deficient mice, CD4+ cells were present
in glomeruli (Fig. 4E), and CD8+ cells
were present in interstitial areas (Fig. 4F), but IFN-
was absent. In IFN-+/+ BM chimeric mice,
IFN- colocalized with numerous CD4+ cells in
the spleen (Fig. 4G). In the kidney (Fig. 4H),
CD4+ cells were observed in glomeruli, and
occasional IFN--positive cells (Fig. 4H, green, possibly
CD8+ cells) were observed in glomeruli, but no
colocalization of IFN- with CD4+ was observed.
IFN- colocalized with CD8+ cells in the
interstitial areas; however, IFN- expression was not observed on any
other cells within the kidney (Fig. 4I). In the spleen of
IFN--/- BM chimeras, no
CD4+ cells expressed IFN-, although some
IFN- expression on non-BM cells (similar to WT mice) was observed
(Fig. 4J). In the kidney (Fig. 4, K and
L), IFN- expression by tubular cells and occasional
glomerular cells was detected, but expression on
CD4+ cells (Fig. 4K) or
CD8+ cells (Fig. 4L) was not observed.
Colocalization of IFN- on macrophages in the kidney was not
detectable in any of the groups of mice (data not shown).
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MHC II expression (Fig. 5A, green) was prominent in
interstitial areas and tubules, and to a lesser extent in glomeruli in
WT mice. Occasional coexpression (Fig. 5A, yellow) with
macrophages was observed. In IFN--deficient mice (Fig. 5B) and in IFN--/- BM chimeric
mice (Fig. 5D), MHC II was very sparsely expressed in
kidneys, and no colocalization with macrophages (Fig. 5, B
and D, red) was observed. In
IFN-+/+ BM chimeric mice (Fig. 5C),
renal MHC II expression was markedly reduced compared with that in WT
mice, but expression was observed on some renal tubular cells and
interstitial cells. Colocalization on macrophages was not apparent.
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WT and sham chimeric mice developed DTH following cutaneous
challenge with sheep globulin. Cutaneous DTH was unaffected in
IFN-+/+ BM chimeric mice but was markedly
reduced in IFN--/- BM chimeric mice and
IFN--deficient mice (Fig. 6A). Serum mouse anti-sheep
globulin Ab titers were equivalent in all groups and were unaffected by
complete or partial IFN- deficiency (Fig. 6B).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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production by BM-derived cells and
resident (non-BM-derived) cells to the development of DTH in kidney and
skin was studied by creating chimeric mice with the capacity for
IFN- production restricted to one of these compartments. The
technique of irradiation and BM transplantation has previously been
successfully used to create mice with genotypically and functionally
distinct BM derived cells (2, 34, 35, 36). In the current
study IFN-+/+ BM chimeric mice were created by
transplanting WT BM into IFN--deficient mice. After 8 wk, these mice
had normal circulating lymphocyte subsets and baseline proteinuria, and
their BM cells were 98% of the transplanted phenotype. Splenic
distribution of lymphocytes and macrophages was normal, with numerous
BM-derived cells expressing IFN-, confirming successful engraftment.
In IFN--/- BM chimeras, created by
transplanting WT mice with IFN--deficient BM, there was no
detectable coexpression of IFN- with CD4+ T
cells in their spleens, indicating depletion of the recipient’s
splenic lymphocytes. Flow cytometric analysis of CD45-positive spleen
cells in transplanted congenic strains confirmed that 90% of splenic
leukocytes expressed the phenotype of the transplanted BM. Studies in
rats show that irradiation totally depletes the small numbers of
resident renal BM-derived MHC II (Ia)-positive cells in 3 days
(37), suggesting that this small resident BM-derived
population has the same radiosensitivity as splenic leukocytes.
Previous studies have demonstrated that IFN- is required for the
development of crescentic GN and cutaneous DTH (11, 13),
both of which are manifestations of Th1-dependent cell-mediated injury.
Studies in CD4+- and
CD8+-deficient mice show a key role for
CD4+ cells in the development of crescentic GN
but demonstrate that injury is independent of
CD8+ cells (38). The results of the
current study demonstrate that while BM cell-derived IFN- alone is
sufficient for cutaneous DTH, contributions from both BM and intrinsic
renal cells are required for full expression of crescentic GN. Crescent
formation, glomerular recruitment of macrophages and
CD4+ T cells, and indices of functional renal
injury (proteinuria and serum creatinine) were significantly reduced in
the absence of IFN- production by either BM cells or intrinsic renal
cells. The development of glomerular and cutaneous DTH was unaffected
by BM transplantation per se, as WT mice transplanted with WT BM (sham
chimeras) developed crescentic GN and skin swelling following Ag
challenge equivalent to those in nontransplanted WT mice.
Previous studies have demonstrated that IL-12 production by
intrinsic renal cells contributes to the development of crescentic GN
(2). The current study provides further evidence of the
contribution Th1 cytokines from intrinsic renal cells to this DTH-like
renal inflammation. Confocal microscopy demonstrated IFN- expression
on tubular cells, interstitial cells, and intrinsic glomerular cells.
However, IFN- production by CD4+ cells and
macrophages in the kidney could not be demonstrated despite prominent
IFN- expression by CD4+ cells in the spleen.
CD8+ cells were the only BM-derived cells that
could be demonstrated to express IFN- at the site of the effector
immune/inflammatory response in the kidney. This suggests that IFN-
from BM-derived cells in secondary lymphoid tissue is important for
development of the Th1 response to the nephritogenic Ag, but that
intrinsic renal cells are an important source of IFN- for the
effector response in the kidney. In contrast, cutaneous DTH is
dependent on BM cell-derived IFN- and is unaffected by the absence
of IFN- from resident cells. These observations indicate a
proinflammatory role for renal parenchymal cells in the development of
renal DTH that does not appear to be shared by keratinocytes in
cutaneous DTH.
In vitro, IFN- has been shown to up-regulate MHC II expression on
mesangial (24, 39, 40), glomerular epithelial (27, 28), renal proximal tubular (24), and endothelial
cells (41). Up-regulation of MHC II allows
murine mesangial cells to stimulate the proliferation of T cells
(42) and to present Ag to T cells (43) in
vitro. Transformed renal tubular epithelial cells can present Ag to MHC
II in vitro (24). In the current in vivo experiment MHC II
was prominently expressed on tubular cells and to a lesser extent on
intraglomerular macrophages in WT mice with crescentic GN. The
expression on other cells within glomeruli was rarely observed,
suggesting that, despite the capacity of mesangial cells to express MHC
II in vitro, they do not necessarily do so in response to inflammatory
stimuli in vivo. These observations in crescentic GN suggest that
proximal tubular cells are more likely than mesangial cells to be
involved in the recognition of Ags in the kidney via MHC II-dependent
mechanisms.
In vivo renal expression of MHC II was markedly reduced (although not
entirely absent) in IFN--deficient mice with GN, indicating an
important, but not exclusive, role for IFN- in MHC II induction.
Residual MHC II expression appeared to be mainly on interstitial cells
and periglomerular cells. Similar findings were reported in
MRL/lpr mice, in which IFN- deficiency was associated
with a dramatic reduction of MHC II expression by tubular cells but
some persistent expression in periglomerular and interstitial cells
(44). In IFN--/- BM chimeric
mice, renal MHC II expression was reduced to a similar extent as in
mice that were totally deficient in IFN-, suggesting that
leukocyte-derived IFN- plays a pivotal role in up-regulating renal
MHC II. Chimeric mice with absent intrinsic renal cell IFN-
production also showed substantially reduced MHC II expression compared
with WT mice with GN, indicating that intrinsic renal cell-derived
IFN- is also required for full expression of MHC II in this disease.
The induction of tubular cell MHC II in the presence of intrinsic renal
cell-derived IFN- may indicate an autocrine role for proximal
tubular cell-derived IFN-. These studies demonstrate that
inflammatory cell and intrinsic renal cell IFN- together play a
major role in the induction of renal MHC II.
The numbers of infiltrating CD4+ T cells and
macrophages were reduced to a greater extent in the
IFN--/- BM chimeras than in
IFN-+/+ BM chimeric mice, suggesting that
systemic or BM cell-derived IFN- plays a dominant role in leukocyte
recruitment. IFN- up-regulates the production of macrophage
inflammatory protein-1
, monocyte chemoattractant protein-1, CSF-1,
and RANTES by mesangial cells (45). In
MRL/Faslpr mice IFN- receptor signaling is
required for the induction of renal CSF-1 and TNF-. The observation
that CSF-1- and TNF--inducing bioactivity could not be detected in
the sera of these mice suggested that the IFN- responsible for these
effects was produced locally within the kidney in this immune
complex-induced model of renal inflammation (46). In the
current studies of a planted Ag-induced, DTH-like model of glomerular
inflammation, it would appear that IFN- from BM-derived cells plays
a predominant role in glomerular and interstitial leukocyte
recruitment. This may be attributed to its important role in driving
Th1 cell development in secondary lymphoid organs.
In conclusion, these studies show that intrinsic renal cells, in
particular tubular cells, produce IFN- in murine crescentic GN.
IFN- from both leukocytes and intrinsic renal cells contributes to
glomerular DTH associated with crescentic GN. IFN- from BM-derived
cells plays a pivotal role in directing the initial Th1-biased immune
response to the nephritogenic Ag, whereas IFN- from resident cells
plays an important role in the effector immune/inflammatory response in
the kidney. Both leukocytes and renal resident cells are important
sources of IFN-, which directs the immune response and mediates
renal injury in crescentic DTH.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Peter G. Tipping, Monash University, Department of Medicine, Monash Medical Center, 246 Clayton Road, Clayton, 3168 Victoria, Australia. E-mail address: peter.tipping{at}med.monash.edu.au
3 Abbreviations used in this paper: GN, glomerulonephritis; BM, bone marrow; c/gcs, cells per glomerular cross-section; DTH, delayed-type hypersensitivity; GBM, glomerular basement membrane; MHC II, MHC class II; WT, wild type.
Received for publication June 21, 2001. Accepted for publication February 20, 2002.
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