1.Copy number variants of ABCF1, IL17REL, and FCGR3A are associated with the risk of gout.
Zheng DONG ; Yuan LI ; Jingru ZHOU ; Shuai JIANG ; Yi WANG ; Yulin CHEN ; Dongbao ZHAO ; Chengde YANG ; Qiaoxia QIAN ; Yanyun MA ; Hongjun HE ; Hengdong JI ; Yajun YANG ; Xiaofeng WANG ; Xia XU ; Yafei PANG ; Hejian ZOU ; Li JIN ; Feng ZHANG ; Jiucun WANG
Protein & Cell 2017;8(6):467-470
2.Eukaryotic expression of human IL17-RD-ECD and generation of its monoclonal antibody.
Xiaojun SUN ; Kunrong MEI ; Yinyin WANG ; Fangli REN ; Yongjing XIA ; Zhijie CHANG
Chinese Journal of Biotechnology 2011;27(10):1507-1512
IL-17 Receptor D (IL-17 RD) is a cytokine receptor that mediates IL-17 signaling and plays an important role in responding to the invasion of extracellular pathogens and many inflammatory and autoimmune diseases such as rheumatoid arthritis. In this study we report the generation of a mouse monoclonal antibody against human IL-17 RD. The recombinant human IL-17RD extracellular domain (hIL-17RD-ECD) was produced in the baculovirus expression system and purified from culture medium of sf9 insect cells. The purified protein was used as a T-dependent antigen to immune Balb/C mice. B cells from the spleen of immunized mice were fused with murine cell SP2/0. Hybridoma cell lines were screened for the production of the monoclonal antibody against hIL-17-RD-ECD using ELISA. A hybridoma cell line 1F8 was found to have a high production of the antibody, which was further confirmed for the specificity by both western blot and ELISA analyses. The monoclonal antibody obtained from hybridoma 1F8 was characterized to be IgG1+Kappa subclass. This study provided a base for the further therapeutic application of the antibody on the autoimmune disease including rheumatoid arthritis.
Animals
;
Antibodies, Monoclonal
;
biosynthesis
;
Baculoviridae
;
Humans
;
Insecta
;
genetics
;
metabolism
;
Mice
;
Mice, Inbred BALB C
;
Receptors, Interleukin-17
;
biosynthesis
;
genetics
;
immunology
;
Recombinant Proteins
;
biosynthesis
;
genetics
;
immunology
3.Expression and clinical significance of IL-17 and IL-17 receptor in ulcerative colitis.
Heng ZHANG ; Bing XIA ; Jin LI ; Qiu ZHAO ; Zhi-tao CHEN ; Rui ZHOU ; Jie WU ;
Journal of Huazhong University of Science and Technology (Medical Sciences) 2016;36(1):37-40
The purpose of this study was to determine the expression levels of IL-17 in serum and IL-17 receptor (IL-17R) in intestinal mucosa tissue in patients with ulcerative colitis (UC) and controls, and evaluate their relationship with disease activity and explore the role of IL-17 in the patho-genesis of UC. A total of 36 Chinese UC patients and 60 healthy controls were enrolled in this study. Serum IL-17 and C-reactive protein (CRP) levels were determined by ELISA and immunonephelometry, respectively. The IL-17R mRNA expression levels were detected by quantitative PCR. Serum IL-17 levels were significantly elevated in UC patients as compared with those in the healthy controls (P<0.05). Among UC patients, serum IL-17 levels were significantly increased in active phase as compared with those in inactive phase (P<0.05), and correlated with CRP levels (r=0.578, P<0.01). IL-17R expression levels were higher in active UC patients than in healthy controls (P<0.05). It was concluded that IL-17 levels were highly expressed in UC, especially in active phase, and correlated with CRP levels in UC patients.
Adult
;
Biomarkers
;
blood
;
C-Reactive Protein
;
metabolism
;
Case-Control Studies
;
Colitis, Ulcerative
;
blood
;
pathology
;
Female
;
Humans
;
Interleukin-17
;
blood
;
Male
;
Middle Aged
;
RNA, Messenger
;
blood
;
Receptors, Interleukin-17
;
genetics
;
metabolism
4.Dexamethasone reduces IL-17 and Tim-3 expression in BALF of asthmatic mice.
Xiao-xia LU ; Karen S MCCOY ; Wei-kun HU ; Jia-li XU ; Hai-qin WANG ; Peng CHEN ; He-bin CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2013;33(4):479-484
This study investigated the expression of interleukin-17 (IL-17) and T cell immunoglobulin mucin and domain-containing molecule-3 (Tim-3) in bronchoalveolar lavage fluid (BALF) of asthmatic mice and the effect of dexamethasone (DEX) on these factors. Thirty-six mice were randomly divided into three groups: normal group, asthmatic group and DEX group. The mouse model of asthma was established by sensitization with ovalbumin in both the asthmatic and DEX groups. The levels of IL-6, IL-10, IL-17 and TGF-β were measured in BALF by enzyme-linked immunesorbent assay (ELISA). The mRNA expression level of Tim-3 was detected by reverse transcription polymerase chain reaction (RT-PCR). The ratio of Tim-3+CD4+ cells to total CD4+ cells in BALF was determined by flow cytometry. Differential inflammatory cells in BALF were detected. The correlations among IL-17, IL-6, IL-10, Tim-3 and inflammatory cells were analyzed. The results showed that the levels of IL-17, IL-6 and Tim-3 were substantially increased and the IL-10 level decreased in BALF in the asthmatic mice, which was significantly reversed by DEX treatment. IL-17 expression was positively correlated with IL-6 and Tim-3 expression and the number of inflammatory cells but negatively with IL-10 expression. These results indicate that the increased expression of IL-17 and Tim-3 in BALF may be implicated in the occurrence and development of asthmatic inflammation; the mechanism by which DEX suppresses asthmatic airway inflammation involves down-regulation of IL-17 and Tim-3 levels.
Animals
;
Asthma
;
drug therapy
;
genetics
;
metabolism
;
Bronchoalveolar Lavage Fluid
;
chemistry
;
Dexamethasone
;
pharmacology
;
Female
;
Gene Expression
;
drug effects
;
genetics
;
Hepatitis A Virus Cellular Receptor 2
;
Interleukin-17
;
genetics
;
metabolism
;
Mice
;
Mice, Inbred BALB C
;
Receptors, Virus
;
genetics
;
metabolism
5.Effects of interleukin-17 on murine pulmonary fibroblast proliferation, transformation and collagen synthesis.
Zhaoxing DONG ; Qingxin KANG ; Wen LEI ; Hong ZHONG ; Wenlin TAI ; Dianhua WANG
Journal of Southern Medical University 2012;32(1):75-79
OBJECTIVETo investigate the effects of interleukin-17 (IL-17) on the proliferation, transformation and collagen synthesis of the lung fibroblasts in mice with bleomycin-induced pulmonary fibrosis.
METHODSIn a mouse model of pulmonary fibrosis established by intratracheal administration of 5 mg/kg bleomycin, the dynamic expressions of IL-17/IL-17 receptor (IL-17R) mRNAs were detected by RT-PCR. At 14 days following bleomycin administration, the pulmonary fibroblasts were isolated, cultured and identified. MTT assay was used to assess the proliferation of the pulmonary fibroblasts in response to IL-17 treatment at different concentrations, and RT-PCR and Western blotting were employed to examine the mRNA and protein expressions of α-smooth muscle actin (α-SMA) and types I and III collagen.
RESULTSIL-17/IL-17R mRNA levels were increased obviously in the pulmonary fibroblasts of rats with pulmonary fibrosis, and the highest expressions occurred at 14 days following bleomycin administration. Exogenous IL-17, at the optimal concentration of 50 ng/ml, significantly promoted the proliferation of the pulmonary fibroblasts in primary culture and obviously increased α-SMA expression and types I and III collagen synthesis in the fibroblasts.
CONCLUSIONIL-17 can promote the proliferation, transformation, and collagen synthesis of the pulmonary fibroblasts from rats with bleomycin-induced pulmonary fibrosis.
Animals ; Bleomycin ; Cell Proliferation ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Epithelial-Mesenchymal Transition ; Fibroblasts ; metabolism ; pathology ; Interleukin-17 ; genetics ; metabolism ; Lung ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Pulmonary Fibrosis ; chemically induced ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Receptors, Interleukin-17 ; genetics ; metabolism
6.Molecular mechanism of acupoint catgut-embedding for experimental colitis in rats.
Xia-Yi ZHANG ; Lin SHEN ; Heng FAN ; Yi LIAO ; Li LIANG
Chinese Acupuncture & Moxibustion 2011;31(10):913-918
OBJECTIVETo explore the effect and its mechanism of acupoint catgut-embedding for experimental colitis in rats.
METHODSEighteen SD rats were randomly divided into a normal control (NC) group, a model (MO) group and a acupoint catgut-embedding (AC) group, 6 rats in each group. Animals in MO group and AC group were treated by trinitro-benzene-sulfonic acid (TNBS) to establish model with colitis. No other treatment was given to the rats in MO group, but acupoint catgut-embedding was implanted at "Shangjuxu" (ST 37), "Tianshu" (ST 25) and "Dachangshu" (BL 25) in the rats in AC group. The symptoms of diarrhea and bloody stool, and histopathology changes in colon were observed 15 days after the treatment. Expression of IL-17, beta2AR, NFkappaBp65 were observed by immunohistochemistry. Expressions of NF-kappaBp65 and beta2AR in splenic lymphocyte were detected by the Western blot method.
RESULTSDiarrhea and mucus bloody purulent stool were soon controlled, and colon mucosa injures were obviously improved in AC group. The NF-kappaBp65 value in splenic lymphocytes of 249.70 +/- 13.66 in MO) group was higher than 86.22 +/- 8.09 in NC group (P < 0.01), and 219.02 +/- 7.42 in AC group was less than that in MO group (P < 0.01). The expression of beta2AR in splenic lymphocytes of 594.97 +/- 173.22 in MO group was less than 957.45 +/- 171.56 in NC group (P < 0.01), and 1335.93 +/- 244.34 in AC group was higher than that in MO group (P < 0.01). The expression of IL-17 in colon mucosa in MO group was increased, while the expression of IL-17 in colon mucosa in AC group was decreased.
CONCLUSIONAcupoint catgut-embedding at Shangjuxu (ST 37), Tianshu (ST 25) and Dachangshu (BL 25) has obviously effect in treating experimental colitis and the mechanism may be related to regulate the expression of IL-17, beta2AR and NF-kappaBp65.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Catgut ; Colitis ; genetics ; metabolism ; therapy ; Disease Models, Animal ; Female ; Gene Expression Regulation ; Humans ; Interleukin-17 ; Male ; NF-kappa B ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; genetics ; metabolism
7.Is Whole Exome Sequencing Clinically Practical in the Management of Pediatric Crohn's Disease?.
Seak Hee OH ; Jiwon BAEK ; Kyung Mo KIM ; Eun Ju LEE ; Yusun JUNG ; Yeoun Joo LEE ; Hyun Seung JIN ; Byong Duk YE ; Suk Kyun YANG ; Jong Keuk LEE ; Eul Ju SEO ; Hyun Taek LIM ; Inchul LEE ; Kyuyoung SONG
Gut and Liver 2015;9(6):767-775
BACKGROUND/AIMS: The aim of this study was to identify the profile of rare variants associated with Crohn's disease (CD) using whole exome sequencing (WES) analysis of Korean children with CD and to evaluate whether genetic profiles could provide information during medical decision making. METHODS: DNA samples from 18 control individuals and 22 patients with infantile, very-early and early onset CD of severe phenotype were used for WES. Genes were filtered using panels of inflammatory bowel disease (IBD)-associated genes and genes of primary immunodeficiency (PID) and monogenic IBD. RESULTS: Eighty-one IBD-associated variants and 35 variants in PID genes were revealed by WES. The most frequently occurring variants were carried by nine (41%) and four (18.2%) CD probands and were ATG16L2 (rs11235604) and IL17REL (rs142430606), respectively. Twenty-four IBD-associated variants and 10 PID variants were predicted to be deleterious and were identified in the heterozygous state. However, their functions were unknown with the exception of a novel p.Q111X variant in XIAP (X chromosome) of a male proband. CONCLUSIONS: The presence of many rare variants of unknown significance limits the clinical applicability of WES for individual CD patients. However, WES in children may be beneficial for distinguishing CD secondary to PID.
Asian Continental Ancestry Group/genetics
;
Carrier Proteins/genetics
;
Child
;
Child, Preschool
;
Crohn Disease/*genetics
;
*Exome
;
Female
;
Genetic Predisposition to Disease
;
*Genetic Variation
;
Humans
;
Immunologic Deficiency Syndromes/genetics
;
Infant
;
Male
;
Phenotype
;
Receptors, Interleukin-17/genetics
;
Republic of Korea
;
Sequence Analysis, DNA/*methods
;
X-Linked Inhibitor of Apoptosis Protein/genetics
8.Advanced glycation end products promote differentiation of CD4(+) T helper cells toward pro-inflammatory response.
Xiao-qun HAN ; Zuo-jiong GONG ; San-qing XU ; Xun LI ; Li-kun WANG ; Shi-min WU ; Jian-hong WU ; Hua-fen YANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(1):10-17
This study investigated the effect of advanced glycation end products (AGEs) on differentiation of naïve CD4(+) T cells and the role of the receptor of AGEs (RAGE) and peroxisome proliferator-activated receptors (PPARs) activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin (BSA) with glucose. Human naïve CD4(+) T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin (sh) RNA knock-down experiment, naïve CD4(+) T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-X(TM) 293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4(+) T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T (Treg) cells was determined by a [(3)H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from naïve CD4(+) T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in naïve CD4(+) T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4(+) T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα PPARγ agonist, PGJ2, inhibited the effect of AGEs on naïve CD4(+) T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4(+) T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity.
Adult
;
Animals
;
Blotting, Western
;
CD4-Positive T-Lymphocytes
;
drug effects
;
metabolism
;
Cattle
;
Cell Differentiation
;
drug effects
;
Cells, Cultured
;
Glucose
;
pharmacology
;
Glycation End Products, Advanced
;
pharmacology
;
HEK293 Cells
;
Humans
;
Interferon-gamma
;
metabolism
;
Interleukin-17
;
metabolism
;
PPAR gamma
;
agonists
;
genetics
;
metabolism
;
Prostaglandin D2
;
analogs & derivatives
;
pharmacology
;
RNA Interference
;
Receptor for Advanced Glycation End Products
;
Receptors, Immunologic
;
genetics
;
metabolism
;
Reverse Transcriptase Polymerase Chain Reaction
;
Serum Albumin, Bovine
;
pharmacology
;
T-Lymphocytes, Regulatory
;
drug effects
;
metabolism
;
Th1 Cells
;
drug effects
;
metabolism
;
Th17 Cells
;
drug effects
;
metabolism