ARTICLE
Auteur(s) :, Jolanta
Myśliwska1,*, K Zorena1, E
Semetkowska-Jurkiewicz2, D Rachoń1, H
Suchanek1, A Myśliwski3
1Department of Immunology, Medical University of
Gdańsk, ul. Dębinki 1, 80-210 Gdańsk, Poland
2Regional Diabetic Center, Medical University of Gdańsk,
Poland
3Department of Histology, Medical University of Gdańsk,
Poland
Insulin-dependent type 1 diabetes mellitus
(type 1 DM) confers an increased risk of various viral,
bacterial and fungal infections [1-3]. Moreover, diabetic
nephropathy (DN) in haemodialysis patients is an independent
predictor of mortality [4]. Although, uncontrolled glycaemia has
commonly been regarded as a causative factor of increased morbidity
and mortality in these patients, this does not represent the whole
picture. The concomitance between the long course of the disease
and the susceptibility to infection leads to the suggestion that
anti-inflammatory and immune-suppressive factors play an essential
role in the pathogenesis of DN. These factors may slow down the
course of diabetic nephropathy through a reduction of the
inflammatory processes, while simultaneously rendering the patient
more susceptible to infection. Interleukin-10 (IL-10) fulfils the
criteria for an anti-inflammatory and immunosuppressive
cytokine.IL-10 has been shown to limit the cascade of
pro-inflammatory cytokines activation [5] and to down-regulate T
cell-mediated immune responses [6]. In accordance with its
anti-inflammatory activity, high concentrations of IL-10 in
critically ill, septic patients protected them from death [7],
while low levels were found in non-survivors [8].The TH2 cytokines
have been documented to as dominating in nephropathies with a
relatively long pre-dialysis or pre-transplantation course such as
idiopathic membranous nephropathy [9], primary IgA nephropathy [10]
and HBV-associated membranous disease [11], all of which are
characterised by an increased production of IL-10 in different in
vitro experimental settings. The role of the TH1 system in
nephropathies is the reverse, as a high expression of TH1 cytokine
transcripts predicts severe glomerular lesions and poor clinical
outcome [12].The studies on experimentally induced, passive,
antiglomerular basement membrane antibody-induced model of
glomerulonephritis in rats showed that systemic treatment with
IL-10 significantly reduced the degree of proteinuria and systemic
inflammation, and attenuated renal injury [13]. Those rats with
mesangial, proliferative glomerulonephritis that received treatment
with IL-10, displayed a lesser degree of histological lesions,
limited cellular proliferation and a lower expression of
inflammatory mediators [14]. When IL-10 was delivered by means of
gene therapy, to mice with naturally occurring renal failure, it
effectively reduced the level of proteinuria and progression to
glomerulosclerosis [15].DN has been regarded as a slowly evolving
disease with a long pre-dialysis period, which suggests that the
course of the disease may be shaped by IL-10.We have not
encountered any reports analysing interleukin-10 in DN patients.
The fragmentary knowledge of the different nephropathies and the
absence of data focusing on diabetic nephropathy, inspired us to
analyse the level of IL-10 in the serum from patients with DN due
to long standing type 1 DM.
Patients and methods
Thirty patients (13 females and 17 males, age 41.76 ± 11.14 years)
with diabetic nephropathy and type 1 DM, were recruited
from The Regional Diabetic Centre at the Medical University in
Gdańsk. Type 1 DM was distinguished from type 2
diabetes by the presence of low BMI, in combination with
hyperglycaemia, ketonuria and a short duration of symptoms such as
polyuria, polydipsia, and weight loss. Their metabolic state was
regularly established by an examination of the fasting glucose
level and glycosylated haemoglobin level (HbA1c).
Moreover, Hb, creatinine, creatinine clearance, total cholesterol,
HDL-cholesterol and triglycerides were monitored. HbA1c
was measured using an immunoturbidometric method involving a
« Unimate 3 » set (Hoffmann-La Roche AG, Germany) with a
normal range of values of 3.0-6.0%. Fasting serum levels of total
cholesterol, HDL-cholesterol and triglycerides were measured with
the enzymatic kits: « Comray Chol », « Comray
HDL-Direct » and « Comray-TG » (P.Z. Comray,
Poland). Systolic and diastolic blood pressures were measured using
an automatic device, with the patient in the supine position after
5 min rest. The mean of all of the blood pressure recordings
measured in each patient during the six months prior to the study
was used. Mean systolic blood pressure below 140 mmHg and mean
diastolic below 90 mmHg were set as threshold values for
normal blood pressure. The urinary albumin excretion rate was
measured in three, timed, overnight urine samples at least twice a
year using an immunoturbidometric assay involving a
« Tina-quant® » (Boehringer Mannheim GmbH,
Germany) kit. Urinary albumin excretion was expressed as the mean
of all 24h collections obtained during the six months prior to our
examination. The serum creatinine level was measured using the
« CREA » assay system (Boehringer Mannheim GmbH, Germany)
with the reference values as follows: for men (< 50 years)
< 115 μmol/L and women < 97 μmol/L. All patients
presented with overt DN, which was characterised as an albumin
excretion rate above 300 mg/24h (macroalbuminuria), in at
least two out of three urine samples during the year, in the
absence of evidence of any other kidney or urinary tract disease.
All patients were receiving angiotensin-converting enzyme (ACE)
inhibitors, four of them in combination with diuretics. Twenty four
patients received HMG-CoA reductase inhibitors. The patients were
not receiving steroids or non-steroidal anti-inflammatory drugs.
The DN patients required insulin treatment at a dose of
20-66 U per day. They had not suffered from any acute or
chronic diseases for three months prior to the examination. The
second group of 30, type 1 DM patients without diabetic
nephropathy (17 females and 13 males, age 37.12 ±
13.30 years) were also recruited from the Regional Diabetic
Centre at the Medical University in Gdańsk. Their metabolic state
was established similarly to that of the group with DN. All
patients were receiving angiotensin-converting enzyme (ACE)
inhibitors. Ten patients received HMG-CoA reductase inhibitors.
They did not receive steroids or non-steroidal anti-inflammatory
drugs. The patients required insulin treatment at a dose of
20-60 U per day. They had not suffered from any acute or
chronic diseases for three month prior to the examination. In both
groups, patients with cardiovascular disease had been excluded from
the study. The control group consisted of 30 healthy subjects
(13 females and 17 males, age 42.5 ± 8.2 years) who did not
suffer from any acute or chronic disease and who had not received
drugs affecting immune system. Written, informed consent was
obtained from all patients and the healthy controls. This study was
approved by the Ethics Committee of The Medical University of
Gdańsk (TKEBN/522/02) and the investigation was carried out in
accordance with the principles of the Declaration of Helsinki as
revised in 1996.
Blood collection
Blood samples were collected between 8.00 and 9.00 after an
overnight fast. The serum was separated from the venous blood
within 30 min and kept frozen at -80 °C, up to three
months prior to analysis. All determinations were done on the same
sample of blood.
Determination of IL-10
Medgenix solid phase ELISA kits (Biosource, Belgium) were used for
the determination of IL-10. The ultra-sensitive assay was applied
(range 0.78-50 pg/mL) to the samples from the DM patients, and
the less sensitive test was used (11-1335 pg/mL) for the
samples from the DN patients. The serum samples were incubated on
microtitre plates coated with capture monoclonal, anti-IL-10
antibodies. Next, the anti-IL-10 HRP conjugate was added, followed
by chromogenic solution TMB. The absorbance was read at 650 nm
on the automated plate reader (Multiscan MCC/340, Labsystems,
Helsinki, Finland). The reference curve was prepared according to
the manufacturer’s recommendations. No significant cross-reaction
was observed between IL-10 and: IL-1, IL-2, IL-3, IL-5, IL-6,
TNF-α, TNF-β, IFN-α, IFN-β, GM-CSF, OSM, MIP-1α, MIF, MCP-1, G-CSF
and RANTES.
Statistical analysis
The results were analysed using the Statistica, Version 6 program
(StatSoft, Pl). The Shapiro-Wilk’s test was used to evaluate the
normality of variables. The differences between the groups of
normally distributed variables were calculated with the unpaired
Student’s t test; with the results presented as arithmetic means ±
SD. For comparison of the skew-distributed variables, the
non-parametric Mann-Whitney U test was applied. The results of the
Mann-Whitney U test are presented as medians (25th and
75th percentiles). The differences in concentration of
IL-10 between patients representing the albuminuria quartiles were
calculated with the ANOVA Kruskal-Wallis test. The Spearman
correlation coefficient was calculated to determine any correlation
between the concentration of IL-10 and albuminuria. The
multivariate linear stepwise regression was applied to assess
independent predictors of albuminuria. The non-normally distributed
values were log-transformed before multivariate regression was
performed. The level of significance was set at p < 0.05 and
two-sided tests were performed as the standard.
Results
Basic clinical characteristics and parameters related to
neuropathy
The parameters related to nephropathy, and the basic clinical
characteristics are presented in tables 1 and 2( Table 1 )( Table 2 ).
The group of 30 patients developed overt nephropathy due to the
long-standing, type 1 DM. They were characterised by
albuminuria in the range of 300 and 3,600 mg/24h (mean: 2064 ±
1030), a creatinine level of 110.5 μmol/L (88.4-150.28) and a
creatinine clearance of 1.59 mL/s (1.49-1.66). They displayed
elevated levels of fasting glucose, HbA1c, and they
required insulin treatment. Systolic and diastolic blood pressures
were within the range of the reference values due to the medical
treatment.
The group of 30, non-DN patients with Type 1 DM were
characterised by an albumin excretion rate < 30 mg/24h, a
creatinine level of 102.5 μmol/L (94.5-109.6) and creatinine
clearance ≥ 1.0 mL/s. This group had elevated levels of
fasting glucose, HbA1c, and required insulin treatment.
Their systolic and diastolic blood pressures, controlled by the
medical treatment, were also within the range of the reference
values (tables 1 and 2).
The basic clinical characteristics of DN patients were similar
to that of Type 1 DM patients. The differences between
values for diabetes duration, BMI, fasting glucose,
HbA1c, Hb, triglycerides, total cholesterol,
HDL-cholesterol and insulin doses were not statistically
significant (table 2).
Table 1 Parameters related to the nephropathy
|
Parameter
|
DN
|
DM
|
Healthy
|
|
patients
|
patients
|
controls
|
|
Number of patients
|
30
|
30
|
30
|
|
Patients with hypertension
|
25
|
20
|
0
|
|
Systolic blood pressure (mmHg)
|
135 (125-140)
|
125 (117−130)
|
120 (115−125)
|
|
Diastolic blood pressure (mmHg)
|
80 (80-85)
|
80 (75-80)
|
79 (70-80)
|
|
Creatinine (μmol/L)
|
110.5 (88.4-150.28)
|
102.5 (94.5-109.6)
|
-
|
|
Creatinine clearance (mL/s)
|
1.59 (1.49-1.66)
|
≥ 1.0
|
-
|
|
Albumin excretion rate (mg/24h) a
|
2064 ± 1030
|
< 30
|
-
|
aThe values are presented as arithmetic means ± SD.
Table 2 Basic clinical parameters
|
Parameter
|
DN
|
DM
|
Healthy
|
|
patients
|
patients
|
controls
|
|
Age (years) a
|
41.76 ± 11.14
|
37.12 ±13.30
|
42.5 ± 8.20
|
|
Duration of the diabetes (years) a
|
22.76 ± 7.30
|
20.66 ± 9.21
|
-
|
|
Duration of the DN (years) a
|
7.50 ± 2.68
|
-
|
-
|
|
6 (20%)
|
5 (17%)
|
7 (23%)
|
|
Body mass index (kg/m2) a
|
24.56 ± 3.00
|
23.44 ± 1.22
|
24.70 ± 2.90
|
|
Fasting glucose (mmol/L) a
|
8.58 ± 1.75
|
8.65 ± 1.75
|
5.20 ± 0.70
|
|
HbA1c (%)
|
7.95 (7.6-8.4)
|
8.2 (6.6-8.8)
|
-
|
|
Hb (g/dL)
|
13.9 (13.6-14.3)
|
12.08 (10.5-12.0)
|
13.76 (13.5-14.6)
|
|
Triglycerides (mmol/L) a
|
1.40 ± 0.66
|
1.23 ± 0.52
|
1.23 ± 0.52
|
|
Total cholesterol (mmol/L)
|
5.34 (4.82-5.88)
|
5.34 (4.87-.16)
|
5.1 (4.5-5.7)
|
|
Cholesterol-HDL (mmol/L)
|
1.49 (1.29-1.83)
|
1.75 (1.47-.16)
|
1.5 (1.33-1.88)
|
|
Dose of insulin (U/day)
|
46 (33-56)
|
38,5 (30-44)
|
-
|
|
n
|
n = 30
|
n = 30
|
n = 30
|
aThe values are presented as arithmetic means ± SD.
Circulating levels of IL-10
The concentration of circulating IL-10 was greatly elevated (10 to
100 times) in 30/30 DM patients with DN (mean
140 pg/mL ± 102), as compared to DM patients without DN, in
whom IL-10 was low, and detectable in only 11/30 patients
(0.79 pg/mL ± 1.24). These low IL-10 serum concentrations in
DM patients without DN were comparable with values in a group of
healthy donors in whom IL-10 was low, and detectable in only
3/30 donors (0.92 pg/mL ± 0.17).
The differences between the three groups (p = 0.04), as well as
between DM patients with DN and DM patients without DN (p =
0.0002), and between DM patients with DN and the control (p =
0.0001) group were statistically significant (table 3)( Table 3 ).
The multivariate stepwise linear regression analysis revealed
that IL-10 was the strongest independent predictor of albuminuria
(β = 1.700; p = 0.0001) in DM patients with DN. Several other
variables also positively influenced the degree of albuminuria, but
with less power. They were: HbA1c (β = 0.481; p =
0.00001), diastolic blood pressure (β = 0.281; p = 0.005) and DN
duration (β = 0.238; p = 0.03). The multivariate regression test
was highly significant (F = 10.81; p = 0.000001) and the model
explained 65% of the variance in the albumin excretion rate.
In the univariate analysis, albuminuria correlated significantly
with the concentration of IL-10 (r = 0.541; p = 0.002) ( (figure 1) ).
Patients in the fourth quartile of albuminuria had a decidedly
higher concentration of IL-10 than those in the quartiles 1 to 3 (F
= 4.6; p = 0.013) ( (figure 2) ).
No correlations between albuminuria and metabolic parameters
were found.
Table 3 Circulating levels of IL-10
|
IL-10
|
DN
|
DM
|
Healthy
|
|
pg/mL
|
patients
|
patients
|
controls
|
|
a
|
121.0 (70-180)
|
0.00 (0.0-1.5)
|
0.92 ± 0.17
|
|
140.0 ± 102
|
0.79 ± 1.24
|
|
|
Statistical significance Mann-Whitney U test
|
|
0.0002
|
0.0001
|
|
Number of patients with detectable IL-10 in serum
|
30/30
|
11/30
|
3/30
|
aThe values are presented as arithmetic means ± SD.
Effect of medical treatment on IL-10 concentration
In our study, according to the results of the multivariate
regression test, neither insulin nor treatment with
angiotensin-converting enzyme (ACE) inhibitors influenced the level
of IL-10 in the patients. The patients receiving a combination of
ACE inhibitors and diuretics also had similar levels of IL-10 as
the patients treated with the ACE inhibitors alone.
Discussion
In the quest for factors that may shape the course of diabetic
nephropathy and influence immune response to microorganisms in DN
patients, we hypothesized that IL-10 may fulfil these criteria. In
line with this assumption, our results have revealed significantly
elevated concentrations of IL-10 in the serum of DN patients. These
concentrations by far exceeded the values present in other
inflammatory diseases such as atopic dermatitis [16] and juvenile
chronic arthritis [17], but were lower in comparison with
circulating IL-10 levels in septic patients [8]. They were however,
similar to those levels found in patients with intractable Grave’s
disease [18].
To our knowledge, this is the first study focusing on the
association between albuminuria and the level of IL-10 in diabetic
nephropathy. Therefore, the answer to the intriguing question about
the causative or protective role of IL-10 in DN can only be
supposed.
IL-10 seems to be able to modulate the natural course of
type 1 DM and of DN at different stages. This effect
however, is non-specific and similar to IL-10-dependent effects in
other nephropathies. Systemic treatment with IL-10 of NOD mice may
delay the onset of the disease or ameliorate its course. This may
be achieved by a transfer of the favouring regulatory response
dendritic cells [19] or by an induction of IL-10-producing
regulatory CD4 + CD25 + cells [20]. The two treatment models are
believed to act by restoring, in the NOD mice, the defective
anti-inflammatory/TH2 immune responses. The decreased in vitro
production of IL-10 in first degree relatives of
type 1 DM patients [21] suggests that faulty
anti-inflammatory/TH2 immune reactions may also underlie human
diabetic disease. Immune cells from recent-onset
type 1 DM patients are able to produce IL-10 in vitro,
but in general, the immune response, at this stage, is biased
towards the TH1 type cytokines [22]. As the disease progresses,
IL-10 may still be found in the serum of some patients but at a
concentration of only a few pg/mL, and with no relation to the
diabetes-associated, immune parameters [23]. These data, together
with the results of our study, suggest that IL-10 probably does not
contribute to the pathology of uncomplicated DM later than during
the pre-diabetic stage.
The immune activation in DN does not seem to be dependent on
antigenic stimulation [24], but appears to be a consequence of
hyperglycaemia. The advanced glycation end products (AGE), induce
inflammatory immune responses that contribute to the growth of the
cortical fibroblasts, collagen synthesis and damage to the proximal
tubular epithelial cells [25, 26]. Mesangial expansion, the
principal glomerular lesion in diabetic nephropathy, has also been
found to be attributable to hyperglycaemia-related inflammation
[27].
It is remarkable that DN, recognized as a devastating disease,
persists for a long time as relatively mild, systemic inflammation
[28]. Given this, our patients with long-lasting DN had relatively
well preserved renal function. This may be due to the protective
role of high concentrations of IL-10.
What is more, the results of our study provide strong evidence
that IL-10 may influence the degree of albuminuria. IL-10 was the
strongest independent predictor of albuminuria in DN patients. In
the univariate analysis, albuminuria significantly correlated with
the concentration of IL-10, and patients in the fourth quartile of
albuminuria had a markedly higher concentration of IL-10 than those
in quartiles 1 to 3. Other variables, such as HbA1c,
diastolic blood pressure and DN duration influenced the degree of
albuminuria with less power. The possible mechanism of action of
IL-10 may be inferred by analogy to other nephropathies. It has
been found that inflammatory/TH1 and anti-inflammatory/TH2 cytokine
transcripts are already weakly expressed in the normal, healthy
kidney [29], and an enhancement of their expression is noticeable
from the very beginning in various nephropathies [12, 30]. Clearly,
the highest expression of IL-10 was found in those patients with
severe proteinuria and extensive glomerular sclerosis, both in
immune-mediated [12, 30] as well as non-immune nephropathies [31].
Thus, a joint induction of IL-10 with a set of counteracting
cytokines from the start of the disease is a general phenomenon
independent of the pathogenesis of the nephropathy. The positive
link between IL-10 and albuminuria presented in this paper
complements the above cited studies. It also however, brings into
question the positive or deleterious effect of IL-10 on the albumin
excretion rate. It may be concluded from an animal model of
nephropathy that IL-10 is not a causative factor of DN, but rather
a regulatory cytokine, opposing the deleterious effect of
inflammatory/TH1 cytokines induced by a poor glyco-metabolic
control. Three studies on various experimentally-induced
glomerulonephritis have demonstrated that treatment with IL-10,
regardless of the route of administration, significantly reduced
the degree of proteinuria [13, 15] and systemic inflammation [13,
14], attenuated glomerular damage and slowed down the progression
to glomerulosclerosis [13, 15]. These papers argue in favour of the
protective role of IL-10 in kidney disorders. The questions remains
as to how to reconcile, the protective role of IL-10 with the
positive correlation found in this study, between the level of
IL-10 and albuminuria, which suggests that IL-10 might be a herald
of DN progression?
The pro-progressive role of IL-10 in DN seems to be executed
indirectly. The production of IL-10 has been documented to be under
the strong control of TGF-β [32], and these two cytokines mutually
regulate each others’ activity [33]. Thus, TGF-β drives the TH1/TH2
balance toward a TH2 immune response with all the positive
consequences cited above. In addition however, TGF-β promotes renal
cell hypertrophy and stimulates extracellular matrix accumulation,
which are two, known hallmarks of diabetic renal disease [34].
Strong expression of TGF-β transcripts has been detected in kidneys
of patients with various nephropathies characterized by marked
proteinuria, but who have preserved renal function. This expression
was related to the degree of renal damage; the plasma and urine
TGF-β 1 protein levels correlated with the amount of protein
excretion [35]. In the long term, the extensive systemic secretion
of IL-10 in DN patients may contribute to the progression of DN and
may be an indicator of marked albuminuria with fairly well
preserved renal function.
To summarise, the DN patients were characterised by a greatly
elevated level of circulating IL-10, which may explain the long
course of the disease with relatively well preserved renal
function. Moreover, the excessive IL-10 production in DN patients
may indirectly contribute towards the progression of DN.
References
1 Eibl N, Spatz M, Fischer GF, et al. Impaired
primary immune response in type-1 diabetes: results from a
controlled vaccination study. Clin Immunol 2002; 103: 249.
2 Chen MK, Wen YS, Chang CC, Lee HS,
Huang MT, Hsiao HC. Deep neck infections in diabetic
patients. Am J Otolaryngol 2000; 21: 169.
3 Moreno R, Zamorano J, Almeria C, et al.
Influence of diabetes mellitus on short- and long-term outcome in
patients with active infective endocarditis. J Heart Valve 2002;
11: 651.
4 Pecoits-Filho R, Barany P, Lindholm B,
Heimburger O, Stenvinkel P. Interleukin-6 is an
independent predictor of mortality in patients starting dialysis
treatment. Nephrol Dial Transplant 2002; 17: 1684.
5 Dokter WH, Koopmans SB, Vellenga E. Effects of
IL-10 and IL-4 on LPS-induced transcription factors (AP-1, NF-IL-6,
NFκB) which are involved in IL-6 regulation. Leukemia 1996; 10:
1308.
6 Korholz D, Banning U, Bonig H, et al. The
role of Interleukin-10 in IL-15-mediated T cell responses. Blood
1997; 90: 4513.
7 Lowe PR, Galley HF, Abdel-Fattah A,
Webster NR. Influence of Interleukin-10 polymorphism on
Interleukin-10 expression and survival in critically ill patients.
Crit Care Med 2003; 31: 34.
8 Yeh FL, Shen HD, Fang RH. Deficient
transforming growth factor β and interleukin-10 responses
contribute to the septic death of burned patients. Burns 2002; 28:
631.
9 Hirayama K, Ebihara I, Yamamoto S, et al.
Predominance of Type-2 immune response in idiopathic membranous
nephropathy. Nephron 2002; 91: 255.
10 De Fijter JW, Daha MR, Schroeijrs WE, Van
Es LA, Van Kooten C. Increased IL-10 production by
stimulated whole blood cultures in primary IgA nephropathy. Clin
Exp Immunol 1998; 111: 429.
11 Lin CY, Lin CC, Chang GJ, King CC. Defect
of cell-mediated immune response against hepatitis B virus: An
indication for pathogenesis of hepatitis-B-virus-associated
membranous nephropathy. Nephron 1997; 76: 176.
12 Lim CS, Zheng S, Kim YS, et al. TH1/TH2
predominance and proinflammatory cytokines determine the
clinicopathological severity of IgA nephropathy. Nephrol Dial
Transplant 2001; 16: 269.
13 Huang XR, Kitching AR, Tipping PG,
Holdsworth SR. Interleukin-10 inhibits macrophage-induced
glomerular injury. J Am Soc Nephrol 2000; 11: 262.
14 Kitching AR, Katerelos M, Mugde SJ,
Tipping PG, Power DA, Holdsworth SR. Interleukin-10
inhibits experimental mesangial proliferative glomerulonephritis.
Clin Exp Immunol 2002; 128: 36.
15 Choi YK, Kim YJ, Park HS, et al.
Suppression of glomerulosclerosis by adenovirus-mediated IL-10
expression in the kidney. Gene Ther 2003; 10: 559.
16 Yoshizawa Y, Nomaguchi H, Izaki S,
Kitamura K. Serum cytokine levels in atopic dermatitis. Clin
Exp Dermatol 2002; 7: 225.
17 Shahin AA, Shaker OG, Hafez N, et al.
Circulating Interleukin-6, soluble Interleukin-2 receptors, tumor
necrosis factor α, and Interleukin-10 levels in juvenile chronic
arthritis: correlations with soft tissue vascularity assessed by
power Doppler sonography. Rheumatol Int 2002; 22: 84.
18 Takeoka K, Watanabe M, Matsuzuka F,
Miyauchi A, Yoshinori I. Increase of serum Interleukin-10
in intractable Grave’s disease. Thyroid 2004; 14:
201.
19 Morin J, Faideau B, Gagnerault MC,
Lepault F, Boitard C, Boudaly S. Passive transfer of
flt-3L-derived dendritic cells delays diabetes development in NOD
mice and associates with early production of interleukin-(IL)-4 and
IL-10 in the spleen of recipient mice. Clin Exp Immunol 2003; 134:
388.
20 Goudy KS, Burkhardt BR, Wasserfall C,
et al. Systemic overexpression of IL-10 induces CD4
+ CD25 + cell populations in vivo and ameliorates
type 1 diabetes in non-obese diabetic mice in a dose-dependent
fashion. J Immunol 2003; 171: 2270.
21 Szelachowska M, Krętowski A, Kinalska I.
Decreased in vitro IL-4 and IL-10 production by peripheral blood in
first degree relatives at high risk of diabetes Type 1. Horm
Metab Res 1998; 30: 526.
22 Kallmann BA, Huther M, Tubes M, et al.
Systemic bias of cytokine production toward cell-mediated immune
regulation in IDDM and toward humoral immunity in Graves’disease.
Diabetes 1997; 46: 237.
23 Hanifi-Moghaddam P, Schloot NC, Kappler S,
Sei-Bler J, Kolb H. An association of autoantibody status
and serum cytokine levels in Type 1 diabetes. Diabetes 2003;
52: 1137.
24 Zerbini G, Bazigalluppi E, Mangili R,
Bosi E, Bonifacio E. revalence of GAD65 and IA-2
auto-antibodies in IDDM patients affected by diabetic nephropathy.
Diabetologia 1997; 40: 485.
25 Morcos M, Sayed AA, Bierhaus A, et al.
Activation of tubular epithelial cells in diabetic nephropathy.
Diabetes 2002; 51: 3532.
26 Jones SC, Saunders HJ, Pollock CA. High
glucose increases growth and collagen synthesis in cultured human
tubulointerstitial cells. Diabetes Med 1999; 16: 932.
27 Wang A, Hascall VC. Hyaluronan structures
synthesized by rat mesangial cells in response to hyperglycemia
induce monocyte adhesion. J Biol Chem 2003; 279: 10279.
28 Saraheimo M, Teppo AM, Forsblom C,
Fagerudd J, Groop PH. Diabetic nephropathy is associated
with low-grade inflammation in Type diabetic patients. Diabetologia
2003; 46: 1402.
29 von Schnakenburg C, Strehlau J, Ehrich JH,
Melk A. uantitative gene expression of TGF-β, IL-10, TNF-α and
Fas ligand in renal cortex and medulla. Nephrol Dial Transplant
2002; 17: 573.
30 Niemir ZI, Ondracek M, Dworacki G, et al.
In situ upregulation of IL-10 reflects the activity of human
glomerulonephritides. Am J Kidney Dis 1998; 32: 80.
31 Rangan GK, Wang Y, Harris DCH. Pharmacologic
modulators of nitric oxide exacerbate tubulointerstitial
inflammation in proteinuric rats. J Am Soc Nephrol 2001; 12:
1696.
32 Maeda H, Kuwahara H, Ichimura Y,
Ohtsuki M, Kurakata S, Shiraishi A. TGF-beta
enhances macrophage ability to produce IL-10 in normal and
tumor-bearing mice. J Immunol 1995; 155: 4926.
33 Cottrez F, Groux H. Regulation of TGF-beta response
during T cell activation is modulated by IL-10. J Immunol 2001;
167: 773.
34 Chen S, Hong SW, Iglesias-de la Cruz MC,
Isono M, Casaretto A, Ziyadeh FN. The key role of
the transforming growth factor-beta system in the pathogenesis of
diabetic nephropathy. Ren Fail 2001; 23: 471.
35 Goumenos DS, Tsakas S, El Nahas AM,
et al. Transforming growth factor-beta (1) in the kidney and
urine of patients with glomerular disease and proteinuria. Nephrol
Dial Transplant 2002; 17: 2145.
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