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*
Department of Immunology, Biogen Inc., Cambridge, MA 02142;
Departments of Medicine, Microbiology-Immunology, and Multipurpose Arthritis Center, Northwestern University Medical School, Chicago, IL 60611
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Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
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Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Critical to the production of Ab against T-dependent Ags is the interaction between CD40L on Th cells and its receptor, CD40, on the cognate B cell. This interaction is essential for germinal center formation, B cell proliferation and differentiation, isotype switching, and generation of B cell memory (reviewed in 10). CD40-CD40L interaction is also important for T cell activation since T cells require costimulatory signals through molecules that are up-regulated upon CD40-CD40L engagement. CD40-CD40L interaction has been shown to be important for several experimentally induced autoimmune diseases, such as collagen-induced arthritis (11), experimental allergic encephalomyelitis (EAE) (12), oophoritis (13), as well as graft-vs-host disease (14, 15), since induction of all of these diseases can be blocked with anti-CD40L treatment at the time of Ag administration. In addition, CD40-CD40L interaction appears to be critical for the production of pathogenic autoantibodies in spontaneous murine lupus. Blocking this interaction, even briefly (for 1 wk) in young, prenephritic SNF1 lupus animals with anti-CD40L therapy produced unexpected long-term benefit, such as increased survival and diminished incidence of severe nephritis at 12 mo of age (16). Similar results were seen in the NZB/NZW lupus model with long-term anti-CD40L therapy (6 mo) (17).
Given the results from these numerous studies, there has been much speculation as to the potential usefulness of a mAb directed against the human CD40L molecule in treatment of autoimmune disease. What is lacking is data that show efficacy of anti-CD40L Ab in established renal disease, both moderate and severe. With this in mind, we designed studies using SNF1 mice that would resemble the stage of disease with which patients with established lupus nephritis could present for initial diagnosis and treatment. In this study, we report the effects of anti-CD40L therapy on animals that began treatment at 5.5 or 7 mo of age, receiving an initial regimen of anti-CD40L Ab for several weeks, followed by monthly dosing for the duration of the study. Anti-CD40L immunotherapy resulted in prolonged survival, decreased autoantibody levels, and diminished proteinuria, indicating an arrest of established disease. Anti-CD40L-treated mice also exhibited reduced renal inflammation, cellular proliferation, vasculitis, and sclerosis/fibrosis, as well as diminished inflammation and fibrosis in the spleen. Lastly, in contrast to 5.5 month-old mice, 7 month-old animals beginning treatment for the first time required more frequent dosing of anti-CD40L in the first 12 wk to establish efficacy, most likely because of components of advanced disease that are not currently understood.
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Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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SWR and NZB mice were purchased from The Jackson Laboratory (Bar Harbor, ME). (SWR x NZB)F1 (SNF1) hybrids were bred in the animal facility at Biogen under conventional barrier conditions. Female SNF1 mice were used for all studies.
Antibodies
The MR1 hybridoma (18), which produces Armenian hamster anti-mouse CD40L Ab, was purchased from the American Type Culture Collection (Rockville, MD). The hybridoma Ha4/8-3.1, an Armenian hamster IgG mAb specific for keyhole limpet hemocyanin, was kindly provided by Dr. Donna Mendrick (Human Genome Sciences Inc., Rockville, MD). Both mAbs were purified from culture supernatant on a protein A Fast Flow column (Pharmacia Biotech, Piscataway, NJ).
Treatment protocols
All injections were given i.p. Each study consisted of a control group that received Ha4/8-3.1 and a treated group that received anti-CD40L mAb. Animals received in the first week 250 µg of Ab on days 1, 3, and 5, then a single dose of 500 µg of mAb once per wk for either 6 or 12 wk as indicated in the text, followed by a single injection of 500 µg monthly until death of the animal or termination of the study. Studies began when animals were either 5.5 mo or 7 mo of age.
ELISA assays
For total Ig and anti-anti-CD40L ELISAs, ELISA plates (Corning Glass Works, Corning, NY) were coated overnight at 4°C with 5 µg/ml of goat anti-mouse IgG+IgM (Jackson ImmunoResearch, West Grove, PA) and anti-CD40L, respectively. After blocking, serial serum dilutions were added, followed by the detection Ab, biotin-conjugated donkey anti-mouse IgG (H+L) (Jackson ImmunoResearch), and streptavidin-horseradish peroxidase (SA-HRP) reagent (Southern Biotech, Birmingham, AL). The developing reaction was stopped by adding 2N sulfuric acid. Plates were read at an OD of 450 nm, and a standard curve was generated using known quantities of purified whole mouse Ig (Jackson ImmunoResearch). Anti-ssDNA and anti-dsDNA ELISAs were performed using NUNC-Immuno Plate MaxiSorp plates (NUNC A/S, Denmark). Plates were coated overnight at 4°C first with 100 µg/ml methylated BSA (Calbiochem Corp, La Jolla, CA), then with 50 µg/ml grade I calf thymus DNA (Sigma, St. Louis, MO). The calf thymus DNA was sheared by sonication and then digested with S1 nuclease before use. For the anti-ssDNA assay, the DNA was boiled for 10 min and chilled on ice before use. After blocking, serial dilutions of serum samples were added and incubated at room temperature for 2 h. Autoantibodies were detected with goat anti-mouse IgG-AP (Sigma) and developed with p-nitrophenyl phosphate (Sigma) in 1 M diethanolamine buffer. Plates were read at an OD of 405 nm, and standard curves were obtained by using known quantities of anti-DNA mAb 205, which is specific for both ss- and dsDNA (2).
Assessment of renal disease
The urine of each mouse was monitored weekly with Albustix (Bayer Corp., Terrytown, NY) to measure proteinuria. Proteinuria level is scored as follows: 0.5+, 15 to 30 mg/dl ; 1+, 30 mg/dl; 2+, 100 mg/dl; 3+, 300 mg/dl; 4+, >2000 mg/dl.
The overall score for histopathologic grading of lupus nephritis is described elsewhere (19, 20) and was based on glomerular, interstitial, and tubular changes. The grades 0 to 4+ are based on percent involvement of the structure being examined (i.e., glomeruli, vessels, etc.). Kidneys without lesions were graded as "0," and all tissue samples were coded and read blind.
Immunohistochemistry
Kidneys and spleens were fixed in 10% buffered formalin and embedded in paraffin. Five-millimeter cryostat sections were baked at 55°C, deparaffined, hydrated in ethanol, and stained with hematoxylin-eosin (H&E) for histologic examination or used for immunohistochemical staining. Briefly, sections were incubated first with a mAb that detects a cytoplasmic protein specific to reticular fibroblasts, ER-TR7 (Serotec, Oxford, UK) for 30 min at room temperature, washed with PBS, incubated with mouse anti-rat IgG (H&L) F(ab)2 for 30 min at room temperature (Jackson ImmunoResearch, West Grove, PA), and visualized using the substrate 3,3' diaminobenzidine (DAB) (Vector Laboratories, Inc., Burlingame, CA). Sections were counterstained with a 25% Wright-Giemsa (Fisher Diagnostics, Pittsburgh, PA) solution. Endogenous peroxidase activity was blocked using 2% hydrogen peroxide in methanol for 20 min before staining with the primary Ab. Photographs were taken on a Zeiss Axioplan photomicroscope at magnifications of x100 and x400.
Statistical analysis
Survival curves were estimated by life-table methodology, and
groups were compared by the Wilcoxon test (21). The proportion of mice
with 
3+ (300 mg/dl) proteinuria was analyzed by a
2 test. Histopathologic renal scores were analyzed by a
Wilcoxon two-sample test. Comparison of autoantibody levels was
analyzed by Student’s t test.
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Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Seven SNF1 mice, beginning at 5.5 mo of age, were
treated in the first week with 250 µg of either anti-CD40L mAb or
control hamster IgG on days 1, 3, and 5 followed by a weekly injection
of 500 µg for 5 consecutive wk, then monthly injections of 500 µg
until death of the animal or termination of the study. By 
10 mo of
age (4.5 mo after the start of treatment), 6 of 7 (85.7%) control
animals had died, whereas no anti-CD40L-treated animals had died
(Fig. 1
A). By 13 mo of
age (7.5 mo after start of treatment) no control animals remained
alive, yet all anti-CD40L-treated animals were alive. In fact,
these animals appeared healthy up to 15.5 mo of age when the study was
terminated and all animals, except one, were euthanized for
histopathology (one mouse died during a kidney biopsy at 13 mo of
age and was not included in the survival timepoints of Figure 1A or statistics beyond 13 mo). As HIg-treated controls
became moribund, the animals were euthanized and their organs removed
for histology. Overall, anti-CD40L-treated mice demonstrated a
survival rate significantly different (p <
0.001) from HIg-treated controls.
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Consistent with the prolonged survival effect of anti-CD40L
therapy described above, this treatment also significantly inhibited
the development of severe nephritis, defined as a proteinuria level of
3+ (p < 0.001 at all
timepoints). As seen in Figure 1
B, only 1 of 7
anti-CD40L-treated animals developed 3+ proteinuria
by 13 mo of age whereas controls rapidly developed 4+
proteinuria within 1 mo after treatment began (although 2 animals had
4+ proteinuria when the study began due to a random
assignment of groups). Remarkably, the proteinuria levels of 6 of 7
(85.7%) anti-CD40L-treated mice declined. This decline began as
early as 3 mo after the start of treatment in some cases and as late as
6 mo in others.
Since anti-CD40L immunotherapy resulted in a decline in
proteinuria, we asked if the severity of glomerulonephritis was also
reduced as compared with controls. For this purpose, H&E-stained kidney
tissue sections were read and scored blind to assess renal morphology
and pathology. An example of normal kidney structure is seen in Figure 2a, a kidney section from an
SWR female mouse, the normal parent of the (SWR x NZB) cross.
Glomeruli are numerous, distinct with patent capillaries, normal
cellularity, and architecture, and tubules are compact and of normal
shape. In comparison, kidney sections from HIg-treated SNF1
mice (Fig. 2b) exhibited severe disruption of kidney
architecture, lesions involving all glomeruli, massive perivascular
lymphoid accumulations, and tubular atrophy or dilation with
proteinaceous casts. The glomeruli in these animals were enlarged and
exhibited hypercellularity with crescents, hyaline deposits effacing
capillary loops, thickening of capillary loops, basement membrane as
well as mesangial thickening, and significant glomerular sclerosis. In
stark contrast to the HIg-treated animals, tissue sections of kidneys
from anti-CD40L-treated animals (Fig. 2c)
revealed that, in general, the overall structural integrity of the
kidneys was intact. Anti-CD40L-treated animals at age 15.5 mo had no to
moderate (0 to 2+) glomerulonephritis, except one mouse
that developed 2 to 3+ disease, and only 3 animals
exhibited rare sclerotic glomeruli. Mouse CLR, which died at 13 mo
of age due to complications from a kidney biopsy procedure, had no
obvious sign of glomerulonephritis in the biopsied tissue (Table I). Furthermore, most animals had no or
only mild interstitial infiltration of mononuclear cells, although 2 of
7 animals had moderate infiltration. A comparison of the overall renal
histopathologic scores for anti-CD40L-treated mice and the controls
shows a significant difference in severity of glomerulonephritis by the
Wilcoxon two-sample test (p < 0.01).
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), and comparison of the difference
in incidence and severity of sclerosis/fibrosis between
anti-CD40L-treated and control mice was found to be significant
(p < 0.01) To further examine the effect of
anti-CD40L therapy on the development of fibrosis, kidney tissue
sections were analyzed with the mAb ER-TR7. This mAb recognizes
reticular fibroblasts and stains the connective tissue of many organs.
(Note: this Ab stains normal connective tissue strongly on frozen
tissue sections. The tissue sections used in this report have been
embedded in paraffin, resulting in a barely detectable staining, except
in instances of damaged and fibrotic tissue.) In a kidney tissue
section of the normal parent, SWR, no staining is detected in the
kidney cortex (Fig. 2d). In HIg-treated animals,
however, this Ab stained intensely the areas surrounding dilated
tubules, interstitial infiltrate, and sclerotic glomeruli (Fig. 2e). Tissue sections of kidneys from
anti-CD40L-treated animals exhibited no ER-TR7 staining above what
was seen for controls (Fig. 2f). Additionally,
examination of H&E-stained kidney sections revealed that, compared with
HIg-treated mice that developed severe vasculitis, the severity of
vasculitis in anti-CD40L-treated animals was significantly
diminished (p < 0.01; Table I). Long-term anti-CD40L therapy beginning at age 5.5 mo reduces splenic inflammation and inhibits development of fibrosis
Since animals in this lupus model develop splenomegaly, splenic
tissue sections were examined to determine whether anti-CD40L
therapy had any effect on inflammation/proliferation in the spleen,
where hyperproliferation of autoantibody-producing B cells occurs.
Normal splenic architecture can be seen in an H&E-stained tissue
section of the normal parent, SWR, (Fig. 3a), where areas of red
pulp and lymphocyte-containing white pulp are clearly discernible. The
spleens of HIg-treated SNF1 mice, however, often had such
severe hyperplasia, accompanied by hyaline degeneration of the central
follicular arterioles, that the typical H&E staining pattern
distinguishing red and white pulp was completely disrupted and
obscured, and there appeared to be a loss of white pulp altogether
(Fig. 3b). There was also evidence of splenic
necrosis in some animals. Splenic tissue sections from animals
receiving anti-CD40L therapy revealed a marked expansion of the
white pulp due to an increased number of follicles and expansion of
what appeared to be dendritic-like cells. Nevertheless, no areas of
necrosis were obvious, and inflammation and lymphoid proliferation were
markedly reduced. In addition, anti-CD40L-treated mice exhibited
only rare, mild incidences of hyaline degeneration of central
arterioles (Fig. 3c).
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d). In the spleens
of HIg-treated mice with active lupus, however, there was extensive
staining with ER-TR7 not only in the red pulp, but extending into the
white pulp as well, which appeared characteristic of periarterial
fibrosis (Fig. 3e). In contrast, in sections from
anti-CD40L-treated mice, there was typical faint staining in the
red pulp, as seen in normal mice, and rare occasions of staining
extending into the white pulp, thus indicating either no or only mild
fibrosis in these animals (Fig. 3f). SNF1 lupus-prone mice beginning immunotherapy at 7 mo of age need a more aggressive anti-CD40L treatment protocol than 5.5 month-old animals
Since long-term immunotherapy with anti-CD40L proved so
successful with animals that began treatment at 5.5 mo of age, this
identical treatment regimen was repeated with older animals, 7 mo old,
that typically have higher proteinuria levels as well as autoantibody
titers. This treatment protocol, however, did not have any beneficial
effect in older mice; anti-CD40L-treated animals developed severe
nephritis and died at the same rate as controls (data not shown). We
reasoned that more frequent anti-CD40L dosing early in therapy
might be necessary given that the autoimmune response in these older
animals is likely more robust than in younger mice. Therefore, 10
animals were given an extended weekly dosing regimen of 500 µg of
either anti-CD40L or HIg for 12 consecutive wk followed by monthly
injections of 500 µg/dose. This aggressive therapy significantly
increased the survival rate of anti-CD40L-treated mice compared
with controls (p = 0.05 by Wilcoxon test). At
10 mo of age (3 mo after start of therapy), only 20% of controls were
alive compared with 80% of treated animals. At 13.5 mo of age the
survival rate was 0% and 40% for control and anti-CD40L-treated
animals, respectively, and at age 15.5 mo when the study was terminated
two anti-CD40L-treated mice remained alive and appeared healthy
(Fig. 4A).
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3+ (300 mg/dl). The
effect of anti-CD40L therapy on proteinuria levels in the 7
month-old mice was not as dramatic as that seen in animals that began
treatment at age 5.5 mo, probably because 6 of 10 animals in the
anti-CD40L-treated group had severe nephritis before the start of
treatment. Regardless, when compared with HIg-treated animals (5 of 10
had severe nephritis before treatment), the proportion of
anti-CD40L-treated mice with 3+ proteinuria differed
significantly (p < 0.001) from controls at
each timepoint analyzed except at 7.5, 8.5, and 10 mo (Fig. 4B). Furthermore, two anti-CD40L-treated animals
exhibited a decline in proteinuria that remained at the diminished
level for several mo (Fig. 4B). Long-term anti-CD40L therapy reduces autoantibody production in SNF1 mice
Autoantibodies, a hallmark of human SLE and the
SNF1 lupus model, are seen in increasing amounts as
SNF1 female mice age. After reaching a peak level, which in
our colony is seen at approximately age 9.5 mo, detectable serum titers
drop dramatically, probably due to immune complex deposition in the
kidneys and other tissues. Serum levels of anti-ssDNA and
anti-dsDNA autoantibodies were determined at regular intervals,
and, regardless of whether mice began anti-CD40L treatment at 5.5
or 7 mo of age, anti-CD40L therapy resulted in an overall reduction
in the mean value of anti-ssDNA and anti-dsDNA autoantibody
detected when compared with HIg-treated controls (Fig. 5). Most animals that began immunotherapy
at age 5.5 mo had detectable autoantibody levels and, whereas the
HIg-treated controls developed increased titers until age 9.5 mo, the
mean values for anti-CD40L-treated mice remained low and in some
cases declined. These differences were significant at the timepoints
indicated in Figure 5 (p < 0.05 at 8.5 mo for
both anti-ssDNA and anti-dsDNA). Mice that began treatment at
age 7 mo also had detectable autoantibody titers at the start of
therapy; however, the mean values of anti-ssDNA and anti-dsDNA
autoantibodies for anti-CD40L-treated mice remained low or declined
compared with controls, which continued to rise until approximately age
9.5 mo. These values differed significantly at certain timepoints (at
8.5 mo, p = 0.06 for anti-ssDNA and
p < 0.05 for anti-dsDNA; at 9.5 mo,
p < 0.05 for anti-dsDNA). Statistical analysis was
not done for those timepoints where there were fewer than 2 control
mice alive.
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). The response, however, was not
persistent and the sporadic nature did not appear to adversely affect
the animals since there was no parallel increase in proteinuria (Fig. 1B), and only 50% of mice had a correlative rise in
autoantibody titers (Fig. 5). On the other hand, several animals that
did not begin anti-CD40L treatment until age 7 mo developed an
anti-anti-CD40L response within 2 mo of therapy, and it was
persistent ) in most cases (Table III). Two of eight
surviving animals 1.5 mo after the start of therapy (age 8.5 mo) had
developed an anti-anti-CD40L response, and this number
increased to four of eight surviving mice at age 9.5 mo. For mice
beginning immunotherapy at age 7 mo, there was a strong correlation
between the animal having severe nephritis at the start of therapy,
development of an anti-anti-CD40L response, and a lack of
long-term survival.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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The hallmarks of interfering with the CD40-CD40L pathway, inhibiting Ig isotype switch and Ab production, have been demonstrated many times in various disease models, primarily, however, in a fashion that inhibited the onset of a humoral immune response. In general most SNF1 animals, without therapeutic intervention, developed significant autoantibody titers as they aged, reaching a peak before disappearance from the periphery due to immune complex deposition in the kidneys and other tissues. Animals that received anti-CD40L immunotherapy, however, maintained consistently low autoantibody titers when compared with controls, often falling below baseline, which was determined just before the start of therapy. Nevertheless, an anti-anti-CD40L response in animals that began treatment at age 7 mo developed rather quickly and was persistent, unlike what was observed in mice that received treatment earlier, at age 5.5 mo, where such a response was sporadic among a few mice and apparently without consequence to disease progression. It is possible that with advanced disease select B cells no longer require a costimulatory signal through CD40, or they may function in a T-independent manner (25), making inhibition of the CD40-CD40L pathway inconsequential. Indeed, CD40L knockout/lpr mice can produce some autoantibodies and develop a markedly delayed and mild form of lupus (26).
Additionally, for mice not receiving anti-CD40L therapy until age 7
mo, increased dosing was necessary to establish efficacy. It was clear
that animals with 3+ nephritis just before dosing did not
benefit from treatment, most likely because of pre-existing kidney
damage that was too severe and irreversible. Possible explanations for
the need of a more aggressive anti-CD40L treatment regimen in
7-month-old animals are an increased level of CD40L expression on Th
cells and a greater number of cells expressing CD40L. Studies have
documented the disregulated expression of CD40L on Th cells in
SNF1 mice (16) as well as in lupus patients (27), whereby
autoreactive Th cells of lupus express an abnormally high level of
CD40L, including T cells taken directly from patients without further
in vitro stimulation. CD40L has also been shown to be expressed on
normal human B cells when stimulated in vitro and, surprisingly, B
cells from lupus patients have been found to exhibit endogenous
hyperexpression of CD40L, reaching the level expressed by activated Th
cells (27, 28). (It should be noted that we have examined both freshly
isolated and mitogen-stimulated purified B cells from SNF1
mice at various ages and have been unable to convincingly detect CD40L
either by flow cytometry or RT-PCR (data not shown)). Interestingly,
previous examination of freshly isolated PBMCs from lupus patients has
shown that patients with the highest level of CD40L expression had
active or end stage renal disease (28). Lastly, with advanced disease,
it is possible that non-T, CD40L-bearing cells, as found in humans,
such as stimulated NK cells (29), vascular endothelial cells, smooth
muscle cells, and macrophages (30) may interact with and activate
CD40+ cells, resulting in an expanded pool of stimulated
lymphoid and non-lymphoid cells.
Together the above data indicate that interrupting CD40-CD40L
interaction not only blocks the initiation and maintenance of the
pathogenic immune response, particularly the humoral arm of the
response, but is also beneficial during the effector phase of disease
when CD40-CD40L interaction takes place in a T/non-B cell or
non-T/non-B cell setting. Indeed, the presence of CD40 on vascular
endothelial cells (31, 32) and on a variety of parenchymal and
nonparenchymal cells in the normal human kidney has already been
established (33). Interestingly, in patients with lupus nephritis, CD40
expression is markedly increased in the kidney along with the presence
of infiltrating CD40L+ mononuclear cells (33). Because
CD40-mediated signals can induce secretion of proinflammatory cytokines
by monocytes, dendritic cells, and fibroblasts (34, 35, 36, 37), it has been
suggested that CD40L+ mononuclear cells may interact with
CD40+ renal target cells to induce or enhance
proinflammatory molecules that contribute to renal inflammation and
damage (33). van Kooten et al. (38) have recently demonstrated that
cross-linking CD40 on human proximal tubular epithelial cells leads to
the production of chemokines IL-8, monocyte chemoattractant protein
(MCP)-1, and RANTES, known inflammatory mediators that may contribute
to the pathway leading to tissue damage and fibrosis. RANTES may be of
particular importance since it is a known chemoattractant for T cells
(39), and IL-2 and IFN-
produced by activated T cells can directly
activate human proximal tubular epithelial cells (40, 41, 42), thus
providing a positive feedback loop for interstitial infiltration. In
fact, Lloyd et al. (43) have used a nephrotoxic serum animal model to
show that MCP-1 is indeed involved in glomerular crescent formation and
interstitial fibrosis and together with RANTES plays a role in the
inflammatory phase of crescentic nephritis. The ability of
anti-CD40L immunotherapy to inhibit the development of fibrosis is
of special significance given that there is a correlation between the
degree of interstitial fibrosis and incidence of chronic renal failure
in patients with glomerular diseases (44, 45). Current studies are
underway to examine CD40, CD40L, and chemokine expression in
SNF1 mice treated with anti-CD40L vs controls.
It has been suggested that anti-CD40L may not be effective after establishment of disease, particularly for Th1-mediated autoimmune diseases (46). Our report demonstrates that long-term immunotherapy with an anti-CD40L mAb provides significant therapeutic benefit to nephritic, autoimmune SNF1 female mice, a lupus model in which the nephritogenic autoantibodies are Th1-dependent (47). Cytokines from CD40L-expressing Th2 cells have also been shown to be necessary for survival of autoimmune B cells and disease progression in lupus (48, 49); thus, our data suggest that preventing CD40-CD40L interaction has consequences for both the Th1 and Th2 subpopulations of T cells. Furthermore, we present the longest survival of SNF1 females ever reported, which is extraordinary at age 15.5 mo. At this age these animals continued to appear in good health with no obvious signs of infection or complications, which is particularly remarkable given that they are autoimmune-prone and were housed in a conventional facility. Importantly, there was no effect on total serum Ig levels (data not shown) indicating there was no general immunosuppression. These data suggest that long-term immunotherapy with anti-CD40L in people may not result in significant detrimental consequences, and overall the data lend promise to the potential use of anti-CD40L immunotherapy to treat human SLE and, possibly, other autoimmune diseases as well.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Susan L. Kalled, Department of Immunology, Biogen Inc., 14 Cambridge Center, Cambridge, MA 02142.
3 Abbreviations used in this paper: SLE, systemic lupus erythematosus; CD40L, CD40 ligand; SNF1, (SWR x NZB)F1; HIg, hamster IgG; H&E, hematoxylin-eosin.
Received for publication July 29, 1997. Accepted for publication November 7, 1997.
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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P I Sidiropoulos and D T Boumpas Lessons learned from anti-CD40L treatment in systemic lupus erythematosus patients Lupus, May 1, 2004; 13(5): 391 - 397. [Abstract] [PDF]
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M. Batten, C. Fletcher, L. G. Ng, J. Groom, J. Wheway, Y. Laabi, X. Xin, P. Schneider, J. Tschopp, C. R. Mackay, et al. TNF Deficiency Fails to Protect BAFF Transgenic Mice against Autoimmunity and Reveals a Predisposition to B Cell Lymphoma J. Immunol., January 15, 2004; 172(2): 812 - 822. [Abstract] [Full Text] [PDF]
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A.-J. Ruth, A. R. Kitching, T. J. Semple, P. G. Tipping, and S. R. Holdsworth Intrinsic Renal Cell Expression of CD40 Directs Th1 Effectors Inducing Experimental Crescentic Glomerulonephritis J. Am. Soc. Nephrol., November 1, 2003; 14(11): 2813 - 2822. [Abstract] [Full Text] [PDF]
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L E Schiffer, N Hussain, X Wang, W Huang, J Sinha, M Ramanujam, and A Davidson Lowering anti-dsDNA antibodies--what's new? Lupus, December 1, 2002; 11(12): 885 - 894. [Abstract] [PDF]
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T. Tanaka, T. Kuroiwa, H. Ikeuchi, F. Ota, Y. Kaneko, K. Ueki, Y. Tsukada, I. B. McInnes, D. T. Boumpas, and Y. Nojima Human Platelets Stimulate Mesangial Cells to Produce Monocyte Chemoattractant Protein-1 via the CD40/CD40 Ligand Pathway and May Amplify Glomerular Injury J. Am. Soc. Nephrol., October 1, 2002; 13(10): 2488 - 2496. [Abstract] [Full Text] [PDF]
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 C. J.J Alderman, P. R Bunyard, B. M Chain, J. C Foreman, D. S Leake, and D. R Katz Effects of oxidised low density lipoprotein on dendritic cells: a possible immunoregulatory component of the atherogenic micro-environment? Cardiovasc Res, September 1, 2002; 55(4): 806 - 819. [Abstract] [Full Text] [PDF]
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C G Katsiari, S-N. Liossis, A M Dimopoulos, D V Charalambopoulos, M Mavrikakis, and P P Sfikakis CD40L overexpression on T cells and monocytes from patients with systemic lupus erythematosus is resistant to calcineurin inhibition Lupus, June 1, 2002; 11(6): 370 - 378. [Abstract] [PDF]
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S. Nierkens, P. van Helden, M. Bol, R. Bleumink, P. van Kooten, S. Ramdien-Murli, L. Boon, and R. Pieters Selective Requirement for CD40-CD154 in Drug-Induced Type 1 Versus Type 2 Responses to Trinitrophenyl-Ovalbumin J. Immunol., April 15, 2002; 168(8): 3747 - 3754. [Abstract] [Full Text] [PDF]
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 I. G. Luzina, S. P. Atamas, C. E. Storrer, L. C. daSilva, G. Kelsoe, J. C. Papadimitriou, and B. S. Handwerger Spontaneous formation of germinal centers in autoimmune mice J. Leukoc. Biol., October 1, 2001; 70(4): 578 - 584. [Abstract] [Full Text] [PDF]
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B H Hahn Lessons in lupus: the mighty mouse Lupus, September 1, 2001; 10(9): 589 - 593. [PDF]
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S. L. Kalled, A. H. Cutler, and L. C. Burkly Apoptosis and Altered Dendritic Cell Homeostasis in Lupus Nephritis Are Limited by Anti-CD154 Treatment J. Immunol., August 1, 2001; 167(3): 1740 - 1747. [Abstract] [Full Text] [PDF]
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V Strand Monoclonal antibodies and other biologic therapies Lupus, March 1, 2001; 10(3): 216 - 221. [Abstract] [PDF]
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D. I. Daikh and D. Wofsy Cutting Edge: Reversal of Murine Lupus Nephritis with CTLA4Ig and Cyclophosphamide J. Immunol., March 1, 2001; 166(5): 2913 - 2916. [Abstract] [Full Text] [PDF]
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J. Zhang-Hoover, A. Sutton, and J. Stein-Streilein CD40/CD40 Ligand Interactions Are Critical for Elicitation of Autoimmune-Mediated Fibrosis in the Lung J. Immunol., March 1, 2001; 166(5): 3556 - 3563. [Abstract] [Full Text] [PDF]
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X. Zhang, D. S. Smith, A. Guth, and L. J. Wysocki A Receptor Presentation Hypothesis for T Cell Help That Recruits Autoreactive B Cells J. Immunol., February 1, 2001; 166(3): 1562 - 1571. [Abstract] [Full Text] [PDF]
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M L Huggins, I Todd, and R J Powell CD40 Ligand--an important target for immunotherapy? Lupus, January 1, 2001; 10(1): 1 - 3. [PDF]
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S L Kalled, A H Cutler, and J L Ferrant Long-term anti-CD154 dosing in nephritic mice is required to maintain survival and inhibit mediators of renal fibrosis Lupus, January 1, 2001; 10(1): 9 - 22. [Abstract] [PDF]
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 A. E. King, R. W. Kelly, H. O. D. Critchley, A. Malmstrom, M. Sennstrom, and R. P. Phipps CD40 Expression in Uterine Tissues: A Key Regulator of Cytokine Expression by Fibroblasts J. Clin. Endocrinol. Metab., January 1, 2001; 86(1): 405 - 412. [Abstract] [Full Text]
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A. M. WOLTMAN, S. DE HAIJ, J. G. BOONSTRA, S. J.P. GOBIN, M. R. DAHA, and C. V. KOOTEN Interleukin-17 and CD40-Ligand Synergistically Enhance Cytokine and Chemokine Production by Renal Epithelial Cells J. Am. Soc. Nephrol., November 1, 2000; 11(11): 2044 - 2055. [Abstract] [Full Text]
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T. Sugiura, Y. Kawaguchi, M. Harigai, K. Takagi, S. Ohta, C. Fukasawa, M. Hara, and N. Kamatani Increased CD40 Expression on Muscle Cells of Polymyositis and Dermatomyositis: Role of CD40-CD40 Ligand Interaction in IL-6, IL-8, IL-15, and Monocyte Chemoattractant Protein-1 Production J. Immunol., June 15, 2000; 164(12): 6593 - 6600. [Abstract] [Full Text] [PDF]
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Z. Liu, K. Geboes, S. Colpaert, L. Overbergh, C. Mathieu, H. Heremans, M. de Boer, L. Boon, G. D'Haens, P. Rutgeerts, et al. Prevention of Experimental Colitis in SCID Mice Reconstituted with CD45RBhigh CD4+ T Cells by Blocking the CD40-CD154 Interactions J. Immunol., June 1, 2000; 164(11): 6005 - 6014. [Abstract] [Full Text] [PDF]
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T. Kuroiwa, R. Schlimgen, G. G. Illei, I. B. McInnes, and D. T. Boumpas Distinct T Cell/Renal Tubular Epithelial Cell Interactions Define Differential Chemokine Production: Implications for Tubulointerstitial Injury in Chronic Glomerulonephritides J. Immunol., March 15, 2000; 164(6): 3323 - 3329. [Abstract] [Full Text] [PDF]
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P. J. Blair, J. L. Riley, D. M. Harlan, R. Abe, D. K. Tadaki, S. C. Hoffmann, L. White, T. Francomano, S. J. Perfetto, A. D. Kirk, et al. Cd40 Ligand (Cd154) Triggers a Short-Term Cd4+ T Cell Activation Response That Results in Secretion of Immunomodulatory Cytokines and Apoptosis J. Exp. Med., February 21, 2000; 191(4): 651 - 660. [Abstract] [Full Text] [PDF]
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 Y.-i. Hwang, M. H. Nahm, D. E. Briles, D. Thomas, and J. M. Purkerson Acquired, but Not Innate, Immune Responses to Streptococcus pneumoniae Are Compromised by Neutralization of CD40L Infect. Immun., February 1, 2000; 68(2): 511 - 517. [Abstract] [Full Text] [PDF]
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 M. L. Stoll and J. Gavalchin Systemic lupus erythematosus--messages from experimental models Rheumatology, January 1, 2000; 39(1): 18 - 27. [Full Text] [PDF]
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F. Mackay, S. A. Woodcock, P. Lawton, C. Ambrose, M. Baetscher, P. Schneider, J. Tschopp, and J. L. Browning Mice Transgenic for Baff Develop Lymphocytic Disorders along with Autoimmune Manifestations J. Exp. Med., December 6, 1999; 190(11): 1697 - 1710. [Abstract] [Full Text] [PDF]
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Z. Liu, S. Colpaert, G. R. D'Haens, A. Kasran, M. d. Boer, P. Rutgeerts, K. Geboes, and J. L. Ceuppens Hyperexpression of CD40 Ligand (CD154) in Inflammatory Bowel Disease and Its Contribution to Pathogenic Cytokine Production J. Immunol., October 1, 1999; 163(7): 4049 - 4057. [Abstract] [Full Text] [PDF]
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T. Kuroiwa, E. G. Lee, C. L. Danning, G. G. Illei, I. B. McInnes, and D. T. Boumpas CD40 Ligand-Activated Human Monocytes Amplify Glomerular Inflammatory Responses Through Soluble and Cell-to-Cell Contact-Dependent Mechanisims J. Immunol., August 15, 1999; 163(4): 2168 - 2175. [Abstract] [Full Text] [PDF]
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 N. S. Kenyon, M. Chatzipetrou, M. Masetti, A. Ranuncoli, M. Oliveira, J. L. Wagner, A. D. Kirk, D. M. Harlan, L. C. Burkly, and C. Ricordi Long-term survival and function of intrahepatic islet allografts in rhesus monkeys treated with humanized anti-CD154 PNAS, July 6, 1999; 96(14): 8132 - 8137. [Abstract] [Full Text] [PDF]
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A. Kaliyaperumal, M. A. Michaels, and S. K. Datta Antigen-Specific Therapy of Murine Lupus Nephritis Using Nucleosomal Peptides: Tolerance Spreading Impairs Pathogenic Function of Autoimmune T and B Cells J. Immunol., May 15, 1999; 162(10): 5775 - 5783. [Abstract] [Full Text] [PDF]
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S K Datta Production of pathogenic antibodies: Cognate interactions between autoimmune T and B cells Lupus, November 1, 1998; 7(9): 591 - 596. [Abstract] [PDF]
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) or HIg (•). An
asterisk (*) denotes points at which the autoantibody levels differed
significantly (p < 0.05 by Student’s
t test); a double asterisk denotes significance,
p = 0.06. Statistical analysis was not performed at
timepoints where the number of surviving control animals was less than
2.