HUMAN IMMUNOLOGY doi: 10.1111/sji.

12057
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Expression of Interleukin-22 in Myasthenia Gravis

S. Zheng*1, C. Dou*1, N. Xin*, J. Wang*, J. Wang*, P. Li*, L. Fu†, X. Shen*, G. Cui*, R. Dong*,
J. Lu* & Y. Zhang*

*Department of Neurology, Affiliated Hospital of
Xuzhou Medical College, Xuzhou, Jiangsu,
China; and †Department of Pathogenic
Organism, Xuzhou Medical College, Xuzhou,
Jiangsu, China
Received 25 February 2013; Accepted in revised
form 18 March 2013
Correspondence to: Y. Zhang, Department of
Neurology, Affiliated Hospital of Xuzhou
Medical College, Xuzhou, Jiangsu, China.
E-mail: zy20037416@163.com
1 significantly decreased in the serum of patients with MG compared with healthy
Shuangshuang Zheng and Changxin Dou
contributed equally to this work.
Abstract
IL-17 and IL-22 are implicated in the pathogenesis of autoimmune diseases. The
roles of IL-22 in the pathophysiology of myasthenia gravis (MG) remain
unsettled. The aim of this study was to investigate the possible relationship
between serum IL-22, IL-17 levels, anti-acetylcholine receptor antibody (anti-
AChR Ab) titres and clinical parameters in patients with MG. The serum IL-22,
IL-17 levels and anti-AChR Ab titres were tested by enzyme-linked immuno-
sorbent assay (ELISA), while the expression of IL-22 and IL-17 mRNAs in
peripheral blood mononuclear cells (PBMC) from healthy and MG subjects were
detected by quantitative real-time PCR (qRT-PCR). Furthermore, PBMC from
12 patients with generalized MG were purified and treated with recombinant
human IL-22 (rhIL-22), the IL-17 levels of supernatant were detected by ELISA.
We found that the IL-17 levels were significantly increased, but IL-22 levels were
controls. Consistantly, a significant decrease in IL-22 mRNA levels and an
increase in IL-17 mRNA levels were detected in PBMC collected from patients
with MG, compared with healthy controls. A negative correlation between IL-22
mRNA in PBMC, serum IL-22 and serum anti-AChR Ab levels was found in
patients with MG. Moreover, in cultured MG PBMC treated with recombinant
human IL-22 (rhIL-22), the IL-17 levels were decreased in a dose-dependent
manner. Our findings indicated a possible role of IL-22 as a protective factor
in MG.

(Treg) cells. Recently, a new T-helper subset, Th17, has

Introduction
The human autoimmune disease myasthenia gravis (MG) is
a T-cell-dependent, antibody-mediated, organ-specific
autoimmune disease, in which the nicotinic acetylcholine
receptor (AChR) at neuromuscular junctions is the major
autoantigen [1]. Patients with MG can be categorized into
two subgroups: (1) patients with purely ocular muscle
weakness (ocular MG); and (2) patients with generalized
muscle weakness, including bulbar and limb muscles
(generalized MG) [2]. Approximately, 80–85% of patients
with generalized MG and 50% of patients with ocular MG
have demonstrable antibodies against AChR. There is a
relationship between the antibody titres and disease in an
individual patient [3, 4].
Although antibodies to AChR are directly responsible
for the destruction of the muscle endplate resulting in both
MG and EAMG, the autoantibody response is T cell
dependent, with CD4+ T cells providing help for B cells to
produce anti-AChR antibodies [5, 6].Activated T cells can
be classified into subsets based on their cytokine production
profiles, that is, T-helper 1 (Th1), Th2 and T-regulatory
been identified and found to have a role in several
autoimmune diseases, such as multiple sclerosis and
systemic lupus erythematosus. These cells secrete a variety
of cytokines, including IL-17, IL-21 and IL-22, with
proinflammatory function [4]. Bai et al. [7] have reported
that IL-17 was a key to autoantibody responses in an
experimental murine autoimmune myasthenia gravis
model. The authors noted that IL-17 elicited higher
antibody responses to the autoantigen AChR. Roche JC
et al. [8] have recently reported that IL-17 concentrations
were higher in generalized MG compared with controls and
correlated with anti-acetylcholinesterase receptor antibody
titres.
Interleukin (IL)-22, a member of IL-10 family of
cytokines, is produced by special immune cell populations,
including CD4+ T cells (Th1, Th17 and Th22 cells), NK
cells, NKT cells and lymphoid tissue–inducer cells [9].
Previous studies have indicated the importance of IL-22 in
host defence against Gram-negative bacterial organisms (in
gut and lung). Recently, there is emerging evidence that
IL-22 is involved in the development and pathogenesis of

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several autoimmune diseases, such as systemic lupus
erythematosus (SLE) [10–14], rheumatoid arthritis (RA)
[15–19], multiple sclerosis (MS) [20–22], Sjögren’s
syndrome (SS) [23, 24] and psoriasis [25, 26]. Therapeutics
targeting IL-22 therefore may have promise for treating
various autoimmune diseases. At present, no clinical data
are available on the expression of IL-22 in patients with
MG; therefore, we measured the serum concentration of
IL-22 and the expression of IL-22 mRNA in PBMCs in the
patients of this disease to determine whether the concen-
tration of this cytokine is linked to clinical parameters and
MG severity.
Materials and methods
Study population. Thirty patients with MG were recruited
prospectively from January 2011 to May 2012 (14 males
and 16 females; mean 41.93  13.48 years, median
39.5 years, range 21–66 years) from the Department of
Neurology at the Affiliated Hospital of Xuzhou Medical
College. The diagnosis of MG was made based on the
following criteria: typical history and signs of fluctuating
weakness of voluntary muscles, presence of serum anti-
acetylcholine receptor antibodies (AChR Ab), definite
clinical improvement on injection of the cholinesterase
inhibitor, edrophonium, and decremental pattern on
repetitive nerve stimulation [27]. The patients were ranked
according to the classification of MG (Osserman) [28]. The
clinical characteristics of the patients are summarized in
Table 1. All patients were classified into one of two groups:
ocular MG (18 patients) or generalized MG (12 patients)
[2]. All patients were seropositive for anti-AChR antibod-
ies. Thymic abnormalities were found in six patients (five
had a thymoma and had undergone a thymectomy, and the
other had a hyperplastic thymus that was diagnosed by
computed tomography). The patients did not have other
autoimmune diseases, ongoing infection and malignancies.
None of the patients had received any immunomodulatory
drugs within the past 3 months. The control group
consisted of 30 healthy subjects with no inflammatory
diseases (14 male and 16 female; 38.33  13.09 years of
age, range 21–65 years). Our study received prior approval
Table 1 General information of study subjects.
Information MG patients
Number of cases 30
Age (Years) 41.93  13.48
Female/Male 16/14
Disease course 6 days–20 years (Mean
34.2 months)
Thymus abnormalities/ 6/5
Thymectomy
MG, myasthenia gravis; HCs, healthy controls.
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Significantly Decreased IL-22 in MG 99
by local ethic committee and informed consent was
obtained from each subject.
Serum samples acquisition. Blood samples were obtained
by venipuncture and immediately centrifuged for 15 min
at 1610 g. Serum samples were stored at 80 °C until
assay. All samples were measured in duplicate and analysed
simultaneously.
RNA extraction. Total RNA was extracted from PBMCs
using Trizol reagent (Invitrogen, USA) according to the
manufacturer’s instructions. PBMCs were separated by
lymphocytes separation medium (Beijing Solarbio Science
& Technology Co., Ltd.) density gradient centrifugation
from heparinized venous peripheral blood. The aqueous
phase was transferred to a fresh tube, and equal volume of
isopropanol was added. After precipitation, samples were
centrifuged at 12,000 9 g for 15 min at 4 °C. The RNA
was washed once with 75% ethanol at 4 °C followed by
recentrifugation. The RNA pellet was air-dried, resus-
pended in nuclease-free water at 500 ng/ul concentration
and stored at 80 °C.
Cytokine measurement by enzyme-linked immunosorbent assay
(ELISA). For the quantification of IL-22 and IL-17 in
human serum samples, Human IL-22 and IL-17 Quanti-
kine Elisa Kit (R&D Systems, Minneapolis, MN) were used
following manufacturer’s guidelines.
Detection of Anti-AChR antibody production by ELISA.
Anti-AChR-IgG responses were measured by indirect
ELISA as described [29, 30]. 96-well flat-bottomed
polystyrene plates were coated with purified AChR
(0.5 lg/ml in 100 ll) overnight at 4 °C, washed with
PBS-T (PBS 0.05% Tween 20) the following day and
blocked with 10% fetal calf serum at room temperature
(RT) for 30 min. Then, washed the plate again with the
same step. Serum (1:100) was incubated at RT for 1.5 h in
a volume of 100 ll. Sera from healthy persons were used
for background control. Each sample was duplicated. After
four washes, HRP-conjugated goat-anti-human IgG
(1:1000) was added and incubated at 37 °C for 1.5 h.
Chromogenic reagent A solution and B solution were
added each 70 ul after four washes, with the seal plate film
sealed the reaction hole. The reaction allowed to develop at
37 °C in the dark for 10 min. Finally, 50 ul termination
Ocular MG Generalized MG (II a and II b) HCs
18 12 30
41  14.69 43.3  11.89 38.33  13.09
10/8 6/6 16/14
6 days–20 years (Mean 3 9. 7 weeks–9 years (Mean –
5 months) 29.2 months)
3/3 3/2 –
100 Significantly Decreased IL-22 in MG
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solution was joined in every reaction hole to terminate the
chromogenic reaction. Plates were read at an OD 490 nm
(OD, optical density) and results expressed as OD
values  standard deviation (SD).
Quantitative real-time polymerase chain reaction (qRT-PCR)
analysis. In accordance with the manufacturer’s instruc-
tions, total RNA from PBMCs was extracted using Trizol
reagent (Invitrogen, USA), according to the method
described above, dissolved in ddH2O processed by DEPC.
Total RNA was reverse-transcribed to cDNA using MMLV
reverse transcriptase (Promega) and looped antisense
primer mix. The synthesized cDNA was amplified by
PCR using the following primers and conditions: Homo
IL-22, sense 5′- CCCTGGCACCCAGCAC-3′, antisense 5′-
GCCGATCCACACGGAGTAC-3′; IL-17, sense 5′-TGTC
ACTGCTACTGCTGCTG-3′, antisense 5′- GTGAGGTG
GATCGGTTGTAG -3′; b-actin, sense 5′- GCTGCCTCC
TTCTCTTGG -3′, antisense 5′- GTGCGGTTGGTGAT
ATAGG -3′. A total 20 ul of real-time PCR system
consists of 2xReal-time PCR Master Mix 10 ul, RNase-free
water 7.6 ul, 0.2 ul forward/reverse primer, 2 ul cDNA
template and 0.4 ul Taq DNA Polymerase. Amplification
and detection were performed as follows: 95 °C, 3 min
degeneration, 95 °C, 30 s, 62 °C, 40 s, a total of 40
cycles. A calibration curve was performed with purified
PCR products of target genes. Melting curve analysis,
obtained by increasing temperature from 60 °C to 95 °C
with a heating rate of 0.1 °C per second and a continuous
fluorescence measurement, revealed a single narrow peak of
suspected fusion temperature. cDNA was amplified by
SYBR Green Universal PCR Master mix (Biorad, Hercules,
CA, USA) in triplicate. All the procedures were strictly
performed as per the instructions of the corresponding kits.
To analyse the expression levels of IL-22 mRNA, we used
the comparative threshold cycle (DDCt) method of relative
quantification with ABI StepOne Plus Detection System.
The results were automatically analysed by the ABI
StepOne Plus instrument and method of 2 DDCt (DCt
represents the difference of threshold cycle value between
the target gene and the inner control; DDCt represents the
difference of DCt between different group) was used to
analyse the mRNA expression.
Cell isolation and culture. PBMC were prepared from
heparinized blood by Ficoll–Hypaque density gradient
centrifugation. Cell cultures were performed as described
previously [31]. In brief, cells were washed twice with PBS to
completely remove the serum. Then, they were suspended in
RPMI 1640 supplemented with 5% calf bovine serum (CBS,
Life Technologies, Inc.) and 1% penicillin–streptomycin
(Life Technologies, Inc.). The cell suspension was adjusted to
a concentration of 2 9 105 cells/ml, and cultured in a 24-
well plastic culture plate in a final volume of 1 ml at 37 °C
in 5% CO 2/95% air. Subsequently, MG and controls
PBMC was cultured with recombinant human IL-22 (rhIL-
22, R&D Systems) (0, 1, 5 or 10 ng/ml) for 96 h in
S. Zheng et al.
combination with anti-CD3 (10 lg/ml) and anti-CD28
(1 lg/ml) (BD Biosciences, San Diego, CA, USA). The
culture supernatant was collected and kept frozen until IL-
17 level were measured.
Statistical analysis. Data are expressed as mean  SD.
Group t test was applied to compare serum level of two
groups. Messenger RNA and anti-AChR Ab levels were
correlated by parametric Pearson correlation analysis. With
a = 0.05 for significant level, P-values less than 0.05 were
considered statistically significant.
Results
Serum IL-17, IL-22 and anti-AChR Ab levels from patients
with MG and healthy controls
We compared serum concentrations of IL-17, IL-22 and
anti-AChR Ab between patients with MG and the normal
control group. The level of IL-17 and anti-AChR Ab was
significantly higher in patients with MG than healthy
controls (Fig. 1A, Fig. 1E, P < 0.001), while the level of
serum IL-22 was significantly lower in patients with MG
(Fig. 1C, P < 0.001); Furthermore, in two types of
patients with MG, the expression level of the three indexes
was also different. There is a significant increase in serum
IL-17 and anti-AChR Ab levels from patients with
generalized MG compared with patients with ocular MG
(Fig. 1B, Fig. 1F, P < 0.01). Higher serum levels of IL-22
was observed in patients with generalized MG compared
with patients with ocular MG (Fig. 1D, P < 0.001).
Measurements of IL-22 and IL-17 mRNA levels in PBMC from
patients with MG and healthy controls
We quantified IL-17 mRNA and IL-22 mRNA levels in
normal control persons and patients with MG by qRT-PCR.
Consistent with the serum level, IL-17 mRNA level was
significantly higher in MG patients than normal persons
(Fig. 2A, P < 0.001), with lower expression in ocular
patients than in generalized ones (Fig. 2B, P < 0.01).
Conversely, as shown in Fig. 2C, patients with MG showed
significantly lower median relative expression of IL-22
mRNA when compared with healthy subjects (Fig. 2C,
P < 0.001). Furthermore, IL-22 mRNA levels were signi-
ficantly decreased in patients with generalized MG with
respect to those with ocular MG (Fig. 2D, P < 0.001).
Correlation between IL-17 mRNA, IL-22 mRNA level in PBMCs
and serum anti-AChR Ab titres
Moreover, a significative correlation between IL-22
mRNA, IL-17 mRNA levels and anti-AChR Ab titres
was found. In all of the 30 patients with MG, IL-17
mRNA level presents a significantly positive correlation
with the expression level of anti-AChR Ab in either all the
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A B

C D
Figure 1 IL-17, IL-22 serum levels and anti-
AChR Ab titres in myasthenia gravis patients
and healthy persons (middle horizontal bars,
mean; each dot, single donor). (A, E) The
levels of IL-17 and anti-AChR Ab were
significantly higher in patients with
myasthenia gravis (MG) than normal persons
(19.31  5.11 pg/ml versus 8.87  2.45 pg/
ml; 0.63  0.08 versus 0.15  0.07); (B, F)
The levels of IL-17 and anti-AChR Ab were
significantly higher in generalized MG
patients than ocular MG patients
(24.11  3.78 pg/ml versus 16.11  2.84 E F
pg/ml; 0.67  0.07 versus 0.61  0.08);
(C) The levels of IL-22 were significantly
lower in patients with MG than normal
persons (19.63  4.59 pg/ml versus 41.95 
6.50 pg/ml). (D) The levels of IL-22 were
significantly lower in patients with generalized
MG than patients with ocular MG
(15.15  2.04 pg/ml versus 22.08  3.59
pg/ml) (**P < 0.01,***P < 0.001).

A B

Figure 2 Detection of IL-17 mRNA and IL-

22 mRNA in PBMCs from patients with D
C
myasthenia gravis and healthy persons by
qRT-PCR. (A, B) IL-17 mRNA levels were
significantly higher in patients with
myasthenia gravis (MG) than normal persons,
with higher expression in generalized patients
than in ocular ones. (C, D) Conversely, the
expression level of IL-22 mRNA decreased
obviously in patients with MG compared with
normal control group, with lower expression in
generalized patients than in ocular ones
(**P < 0.01, ***P < 0.001).

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102 Significantly Decreased IL-22 in MG S. Zheng et al.
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A D

B E

C F

Figure 3 Correlation between IL-17,IL-22 mRNA expression and anti-AChR Ab titres. (A, B, C) IL-17 mRNA level presents a significantly positive
correlation with the expression level of anti-AChR-IgG in either all the myasthenia gravis (MG) subjects or the different types of patients with MG, the
correlation coefficient is 0.536 (P < 0.01), 0.551 (ocular MG, P < 0.05) and 0.619 (generalized MG, P < 0.05) respectively; (D) IL-22 mRNA level
presents a significantly negative correlation with the expression level of anti-AChR Ab in all the patients with MG, the correlation coefficient is 0.566
(P < 0.01). (E, F) The IL-22 mRNA and anti-AChR Ab also exist negative correlation in the different types of patients with MG,and the correlation
coefficient is 0.549(ocular MG, P < 0.05) and 0.583 (generalized MG, P < 0.05) separately.

MG subjects or the different types of patients with MG
(Fig. 3A–C); On the contrary, IL-22 mRNA level presents
a significantly negative correlation with the expression
level of anti-AChR Ab in all the patients with MG
(Fig. 3D). Moreover, the IL-22 mRNA and anti-AChR Ab
also present negative correlation in the different types of
patients with MG (Fig. 3E, F).
Correlation between serum IL-17, IL-22 levels and anti-AChR
Ab titres
Meantime, a significative correlation between IL-22, IL-17
levels in serum and anti-AChR Ab titres was found. In the
30 patients with MG, IL-17 serum level presents a
significantly positive correlation with the expression level

Scandinavian Journal of Immunology, 2013, 78, 98–107

S. Zheng et al. Significantly Decreased IL-22 in MG 103
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A D

B E

C F

Figure 4 Correlation between serum IL-17 and anti-AChR Ab titres (A, B, C), serum IL22 and anti-AChR Ab titres (D, E, F), serum IL-17 level presents an
positive correlation with the expression level of anti-AChR-IgG in either all the myasthenia gravis (MG) subjects or the different types of patients with MG,
the correlation coefficient is 0.538 (Fig. 4A, P < 0.01), 0.510 (Fig. 4B, ocular MG, P < 0.05) and 0.589 (Fig. 4C, generalized MG, P < 0.05) respectively.
Serum IL-22 level presents a significantly negative correlation with the expression level of anti-AChR Ab in all the patients with MG, the correlation
coefficient is 0.572 (Fig. 4D, P < 0.001) Moreover, the IL-22 serum level and anti-AChR Ab also exist negative correlation in the different types of patients
with MG, and the correlation coefficient is 0.582 (Fig. 4E, ocular MG P < 0.05) and 0.629 (Fig. 4F, generalized MG P < 0.05) respectively.

of anti-AChR Ab in either all the MG subjects or the
different types of patients with MG (Fig. 4A–C); On the
contrary, IL-22 serum level presents a significantly negative
correlation with the expression level of anti-AChR Ab in all
the patients with MG (Fig. 4D). Moreover, the IL-22 serum
level and anti-AChR Ab also exist negative correlation in the
different types of patients with MG (Fig. 4E, F).
Correlation analysis between IL-17 and IL-22 level in patients
with MG
A significantly negative correlation between IL-22 and IL-
17 levels in either PBMCs or serum of MG subjects was
found. In all of the 30 patients with MG, IL-17 mRNA
level presents a significantly negative correlation with the

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104 Significantly Decreased IL-22 in MG S. Zheng et al.
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A D

B E

C F

Figure 5 Correlation between IL-17 and IL-22 expression either in PBMCs or in the serum of patients with myasthenia gravis (MG). (A, B, C) In the
PBMCs, IL-17 mRNA presents a significantly negative correlation with the expression level of IL-22 mRNA in either all the MG subjects or the different
types of patients with MG, the correlation coefficient is 0.884 (P < 0.001), 0.668 (ocular MG, P = 0.002) and 0.613 (generalized MG, P < 0.05)
respectively; (D, E, F) In the serum, IL-17 level also exists a significantly negative correlation with the expression level of IL-22 in all the patients with
MG, as well as in the different types of patients with MG, the correlation coefficient is 0.805 (P < 0.001), 0.499 (ocular MG, P < 0.05) and 0.720
(generalized MG, P < 0.01) separately.

expression level of IL-22 mRNA in either all the MG
subjects or the different types of patients with MG
(Fig. 5A–C); Similarly, the expression level of IL-17 in the
serum of patients with MG presents a significantly negative
correlation with the expression level of IL-22, as well as in
the different types of patients with MG (Fig. 5D–F).
Downregulated expression of IL-17 in PBMC by stimulation
with rhIL-22
We investigated the effect of different concentrations of
rhIL-22 on IL-17 production from MG and control PBMC.
After stimulation of PBMC with rhIL-22, the IL-17

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A B

Figure 6 Effect of recombinant human IL-22

(rhIL-22) on interleukin (IL)-17 release by
anti-CD3 and anti-CD28-stimulated PBMC
from patients with generalized myasthenia C D
gravis and healthy controls. PBMC were
cultured at a concentration of 2 9 105 cells
per well with medium, anti-CD3, anti-CD28
and rhIL-22 under the conditions described in
the Materials and methods section. After 96 h
of treatment, IL-17 expression in culture
supernatant was measured by ELISA (A, B).
Cell viability was assessed by the trypan blue
dye exclusion method and expressed as a
percentage with the formula 100 9 (number
of viable cells/number of both viable and dead
cells (C, D). (*P < 0.05,**P < 0.01, ***P <
0.001).

expression in culture supernatant was determined by
ELISA. As shown in Figure 6A,when PBMC from patients
with MG were treated with rhIL-22, the levels of IL-17
decreased in a dose-dependent manner. A significant
decrease of IL-17 in patient with MG was detected in
1 ng/ml rhIL-22 treatment group. When PBMC from
healthy controls were treated with rhIL-22, modest
decrease of IL-17 in culture supernatant was observed in
10 ng/ml rhIL-22 treatment group. Cytotoxic effects on
PBMC by rhIL-22 at experimental concentrations were not
observed (Fig. 6C, D).
Discussion
In our study, we found that the expression of IL-17 mRNA
in PBMC and serum IL-17 levels was higher in patients with
MG than in controls, with the results consistent with
previous study [8]. We also found that IL-17 mRNA and
protein level in patients with generalized MG was higher
than that of patients with ocular MG. Moreover, we analysed
the relationships of IL-17 and anti-AChR Ab levels. We
have arrived at the conclusion that IL-17 expression was
positively correlated with anti-AChR Ab titres. Integrated
above all the evidence, we can conclude an important role for
IL-17 in the pathogenesis of patients with MG.
About IL-22, on the one hand, our results showed that
IL-22 mRNA in PBMC and IL-22 levels in serum were
significantly decreased in patients with MG compared with
normal controls; on the other hand, significant difference
was also found between ocular MG and generalized MG.
Moreover, correlation analysis between IL-22 mRNA levels
and anti-AChR Ab titres showed negative association. The
decreased serum IL-22 level in MG indicated that IL-22
might be protective for MG.
Unregulated T-cell responses or overwhelming cyto-
kines secreted by T cells and other cell sources are
associated with many autoimmune diseases such as SLE,
RA, MS, SS and psoriasis. IL-22 and other Th17 cytokines
have been most commonly described as a pro-inflammatory
cytokine due to its expression in lesions of patients with
chronic inflammatory diseases and its induction of pro-
inflammatory cytokines such as IL-6, IL-8 and TNF [32].
However, other data suggest that IL-22 has a less direct
inflammatory role and instead induces expression of genes
associated with antimicrobial defence and cellular differ-
entiation [26]. Moreover, a protective role of IL-22 has been
described in inflammatory bowel disease [33, 34], exper-
imental hepatitis [32] and experimental autoimmune
myocarditis [35]. These discrepancies may be explained
by the dual role of IL-22 in inflammation, for example, IL-
22 plays a pro-inflammatory role in the onset of ovalbumin-
induced model of allergic asthma in mice; in contrast,
IL-22 plays a regulatory role in established asthma and
participates to the resolution of inflammation [36]. In
addition, the role of IL-22 in inflammation differs
depending on the specific tissue [34]. IL-22 contributes

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106 Significantly Decreased IL-22 in MG
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to the regulation of hepatitis [37], whereas dermal
inflammation is mediated by this cytokine [26]. Studies
found that serum IL-22 is upregulated in several autoim-
mune diseases such as RA [17], MS [22], SS [24] and
psoriasis [25], but decreased serum and plasma IL-22 levels
were observed in SLE patients by Pan et al. and Cheng
et al., respectively [10, 11].The reason for this phenomenon
is not known. Ziesche et al. [12] have previously demon-
strated that glucocorticoid dexamethasone (DEX) can
suppress IL-22 production of plasma and PBMCs in the
context of acute bacterial infections, this result may
partially explain the decreased levels of serum IL-22 in
SLE patients. However, McKinley et al. [9] reported that
IL-22 production was not sensitive to DEX treatment at
any doses tested. Our study found that plasma IL-22 level
was downregulated in patients with MG. However, our
patients had received no DEX within the past 3 months,
which suggesting that downregulated IL-22 in patients
with MG have little relation to DEX. AS far as we know,
both SLE and MG are antibody-mediated autoimmune
diseases, they might share similar underlying mechanism
required for disease progression. Therefore, further studies
are needed to explore the potential mechanisms of IL-22 in
MG and other autoimmune diseases.
Previous studies had already shown a cross-talk between
IL-17A and IL-22 production [38, 39]. Sonnenberg and
co-workers [40] showed in vitro that IL-17A could suppress
IL-22 expression from Th17 cells in a dose-dependent manner.
In vivo, they found an increase of IL-22 in the lung of mice
deficient in IL-17A after bleomycin injury. Ke et al. [41]
showed that IL-22 treatment in experimental autoimmune
uveitis changed the function of Ag-primed CD11b+ APCs,
which then bestowed autoreactive pathogenic Th17 cells
with regulatory activities. Similarly, Besnard et al. [36]
showed that IL-17A production is abolished in the lung by
exogenous IL-22 treatment during antigen challenge,
which protects mice from lung inflammation. Collectively,
these observations suggest a reciprocal regulation of
IL-17A and IL-22. However, the underlying molecular
mechanisms are unknown. In our study, we found a
negative correlation between IL-17 and IL-22 expression in
patients with MG, suggesting that this two cytokines play
opposite role in this disease. Moreover, in agreement with
previous studies [36, 41], we found that IL-17 production
by activated MG PBMC is inhibited in the presence of
rhIL-22, but IL-17 production produced by controls’
PBMC had a slight decline in the presence of high-dose
rhIL-22. This result suggested that PBMC cells from
patient with MG are more sensitive to rhIL-22 treatment
in comparison with PBMC cells from healthy control.
In summary, the decreased serum IL-22 level and IL-22
mRNA in PBMC from persons suffering of MG indicated
that IL-22 might be protective for MG. Given that IL-22 is a
potent anti-inflammatory cytokine in MG, an important
question is whether IL-22 can be a therapeutic target. In
S. Zheng et al.
MG, a variety of cytokines, including IFN-c, IL-1 and IL-
17, are thought to play a pathogenic role. The relative
contribution of these inflammatory cytokines to MG can
differ. In this context, the possibility that supplementing
IL-22 impedes the progression of MG should be explored in
animal models of MG. For example, development of
recombinant IL-22 engineered to last long in vivo, discovery
of small chemical compounds that mimic IL-22 signalling
for immunosuppression will contribute to the development
of novel therapeutic approaches to manage MG. Further
studies are necessary to establish the pathophysiologic role
of IL-22 in MG. As the decreased IL-22 mRNA levels were
found in patients with generalized MG with respect to those
with ocular MG, IL-22 could be considered a potential
marker of the severity of disease. Moreover, we did found
association of IL-22 mRNA levels with anti-AChR Ab, this
is a noteworthy finding, although this may be explained by
the small sample size of this study. Therefore, prospective
cohort studies with large sample size are needed.
Acknowledgment
We would like to thank all patients and normal control
persons for their collaboration. This work was supported by
National Nature Science Foundation of China (81072465),
Natural Science Fund of the Educational Committee of
Jiangsu Province (10KJD320003), Special foundation of
president of the Xuzhou Medical College (2010KJZ01),
Key medical talents fund of Jiangsu Province (H201130),
Jiangsu Province ordinary university postgraduate research
innovation fund (CXLX11_0734).
Disclosures
The authors have no financial conflict of interest.
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