December 2010
Volume 51, Issue 12
Free
Clinical and Epidemiologic Research  |   December 2010
Association of Interleukin-1β (IL1B) Polymorphisms with Graves' Ophthalmopathy in Taiwan Chinese Patients
Author Affiliations & Notes
  • Yu-Huei Liu
    From the Departments of Medical Genetics and Medical Research and
    the School of Chinese Medicine,
  • Rong-Hsing Chen
    the School of Post-Baccalaureate Chinese Medicine, and
  • Hsin-Hung Wu
    the Department of Business Administration, National Changhua University of Education, Changhua, Taiwan; and
  • Wen-Ling Liao
    From the Departments of Medical Genetics and Medical Research and
    the School of Chinese Medicine,
  • Wen-Chi Chen
    the Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan;
  • Yuhsin Tsai
    the School of Chinese Medicine,
  • Chang-Hai Tsai
    Pediatrics, China Medical University Hospital, Taichung, Taiwan;
    the Departments of Biotechnology,
    Biotechnology and Bioinformatics, and
  • Lei Wan
    From the Departments of Medical Genetics and Medical Research and
    the School of Chinese Medicine,
    Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan.
  • Fuu-Jen Tsai
    From the Departments of Medical Genetics and Medical Research and
    Pediatrics, China Medical University Hospital, Taichung, Taiwan;
    the School of Chinese Medicine,
    the School of Post-Baccalaureate Chinese Medicine, and
    the Departments of Biotechnology,
    Biotechnology and Bioinformatics, and
  • *Each of the following is a corresponding author: Lei Wan, Genetic Center, China Medical University Hospital, No.2 Yuh-Der Road, 404 Taichung, Taiwan; leiwan@mail.cmuh.org.tw. Fuu-Jen Tsai, Genetic Center, China Medical University Hospital, No.2 Yuh-Der Road, 404 Taichung, Taiwan; d0704@mail.cmuh.org.tw
Investigative Ophthalmology & Visual Science December 2010, Vol.51, 6238-6246. doi:10.1167/iovs.09-4965
  • Views
  • PDF
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Yu-Huei Liu, Rong-Hsing Chen, Hsin-Hung Wu, Wen-Ling Liao, Wen-Chi Chen, Yuhsin Tsai, Chang-Hai Tsai, Lei Wan, Fuu-Jen Tsai; Association of Interleukin-1β (IL1B) Polymorphisms with Graves' Ophthalmopathy in Taiwan Chinese Patients. Invest. Ophthalmol. Vis. Sci. 2010;51(12):6238-6246. doi: 10.1167/iovs.09-4965.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose.: To evaluate whether variations in the IL1B gene could be associated with Graves' ophthalmopathy (GO) in patients with Graves' disease (GD).

Method.: This case–control study included 471 Taiwan Chinese patients with GD (200 with GO and 271 without GO) and 160 healthy volunteers. Eight single-nucleotide polymorphisms (SNPs) in IL1B were genotyped with an allele-specific extension and ligation assay.

Results.: In the IL1B SNPs examined, the C allele of rs1143634 was associated with GD, whereas the T/T genotype of the SNPs rs1143634 and rs16944 were less associated with the disease. The A/A genotype of the SNPs rs3917368 and rs1143643, which had the strongest interaction, may increase the risk of GO (P = 0.024 and P = 0.017, respectively). Several GD susceptibility and insusceptibility IL1B haplotypes have been identified, and the Ht4-GCGCCTCC haplotype, composed of eight SNPs and associated with low circulating IL1β levels, may be protective against the development of GO (P = 0.025). Moreover, that the GO-susceptible genotype was associated with lower plasma IL1β concentrations implies that the origin of GO may go beyond the IL1B polymorphism-associated elevation of circulating IL1β.

Conclusions.: The data for IL1B polymorphisms and the association of GD and GO with plasma IL1β levels show that IL1B polymorphisms may be associated with the development of GD and GO.

Graves' disease (GD), with or without Graves' ophthalmopathy (GO), is an autoimmune disease characterized by hyperthyroidism, diffuse goiter, thyroid-specific autoantibodies, and dermopathy due to circulating autoantibodies. 1 GO is the most common extrathyroid manifestation of GD and affects 25% to 50% of GD patients. 2 5 Approximately 28% of patients with GO present as severe cases, with restricted mobility, diplopia, keratopathy, and optic neuropathy. 6,7 Several genes have been reported that promote the development of GO, including human leukocyte antigen (HLA) class I and class II molecules. 8 For example, the +49G allele of cytotoxic T-lymphocyte-associated antigen-4 (CTLA4) confers genetic susceptibility to GO, although meta-analyses did not support this finding. CT60A/G of CTLA4 is one of the GD-associated polymorphisms that await further studies that examine their association with GO. In addition, polymorphisms in several immunomodulatory genes, such as the intron 1 (CA) repeat in interferon-γ (IFNG); G238A, C863A, and T1031C in tumor necrosis factor-α (TNFA); and A1405G in intracellular adhesion molecule-1 (ICAM1), have been reported to increase susceptibility to GO. 8 The combination of specific alleles among these genes would make the patient susceptible to GO. 
Interleukin-1 beta (IL1B), a proinflammatory cytokine expressed by activated macrophages and several other types of cells, is thought to play a crucial role in the pathogenesis of autoimmune diseases. 9,10 IL1β was initially known as one of the lymphocyte activating factors (LAFs), owing to its role in the induction of T-cell proliferation and maturation. 9,10 Recent studies have demonstrated that IL1β released by macrophages and fibroblasts can induce adipogenesis and accumulation of glycosaminoglycans (GAGs) and prostaglandin E2 (PGE2), which may result in the development of GO. 8,11,12 One study revealed that the single-nucleotide polymorphism (SNP) −511C of IL1B is associated with GO, whereas others did not. 13 15 Although the connection between IL1B polymorphism and GO remains controversial, several studies have demonstrated that polymorphisms of IL1B may correlate with IL1β expression in other diseases. 16 18 In addition, IL1β promotes the accumulation of GAGs through the upregulation of hyaluronan synthesis and accumulation of PGE2, stimulates adipogenesis, and hyperinduces the expression of interleukin (IL)-6 and -8 and macrophage chemoattractant protein (MCP)-1 in orbital fibroblasts derived from healthy individuals and patients with GO. 8,19 23 Moreover, the polymorphisms of the IL1 family are involved in several autoimmune diseases such as systemic lupus erythematosus (SLE), 24 26 rheumatoid arthritis, 27,28 autoimmune hemolytic anemia, 29 and GD. 30 These reports support that IL1B is a potential candidate gene in the development of GO. 
Although previous reports have suggested that IL1B polymorphisms and expression lead to autoimmune diseases, 24 30 the genetic role of IL1B in GO remains to be elucidated. In the present study, we investigated SNPs in IL1B that may be protective against or causative of GO in Taiwan Chinese patients with GD. 
Methods
Patients and Healthy Individuals
A group of 484 patients with a confirmed diagnosis of GD and a control group of 160 healthy volunteers at China Medical University Hospital in Taiwan were enrolled and actively observed. All individuals in this study provided informed consent, as approved by the ethics committee of China Medical University Hospital and in accordance with the guidelines in the Declaration of Helsinki. 
Patients.
Diagnosis of GD was based on the typical clinical features of hyperthyroidism: diffuse enlargement of the thyroid gland, increased free thyroxine or triiodothyronine levels, suppressed thyroid-stimulating hormone levels, positive thyrotrophin-receptor autoantibodies, and the presence (or absence) of anti-thyroid peroxidase (anti-TPO) antibodies or antithyroglobulin antibodies. Information regarding sex, age at onset of GD, treatment of hyperthyroidism, personal history of cigarette smoking, history of systemic diseases, and family history of autoimmune thyroid disease was obtained. The inclusion criteria were (1) meeting the diagnostic criteria of GD at the time of examination; (2) being willing to participate and capable of giving informed consent; and (3) being a self-reported nonaboriginal Taiwanese with no parent or grandparent having an aboriginal background. The exclusion criteria were (1) being unable to understand or give informed consent or (2) being pregnant or having given birth within 1 year, to exclude the possibility of including subjects with postpartum thyroiditis. Patients with GO (GD/GO) were identified according to the following criteria: Normal upper eyelid position was 1.5 mm below the superior limbus, and normal lower eyelid position was at the level of the inferior limbus in primary gaze. Proptosis was measured by a Hertel exophthalmometer and was defined as the anteroposterior protrusion of the globe >19 mm from the lateral orbital rim in either eye or any discrepancy in the degree of protrusion of the two eyes by >1 mm. All individuals classified as affected were interviewed and examined by experienced clinicians. A full medical record review was conducted to obtain demographics (age and sex); history of tobacco use; recurrence of GD (patients with GD who have accepted medical treatment); and progression (patients with ongoing GD), treatment, and clinical features of the condition. 
Healthy Individuals.
The healthy group was matched for sex according to the female predominance of GD, including 32 men (20.0%) and 128 women (80.0%). Age was significantly different between the groups of healthy volunteers (27.4 ± 6.4 years) and patients with GD (39.9 ± 12.2 years; P = 1.028 × 10−34). 
All blood samples were collected from consenting individuals by venipuncture for subsequent genomic DNA isolation. 
SNP Selection
IL1B SNP genotype information was downloaded in December 2008 from the HapMap CHB+JPT population. HapMap genotypes were analyzed in Haploview (Haploview software (http://www.broad.mit.edu/mpg/haploview/ provided in the public domain by The Broad Institute, Massachusetts Institute of Technology, Cambridge, MA), and Tag SNPs were selected by using the Tagger function and applying the following additional criteria: (1) a threshold minor allele frequency (MAF) in the HapMap CHB+JPT population of 0.10 for tag SNPs and (2) a genotyping score (Illumina, Inc., San Diego, CA) more than or equal to 0.6, as recommended by the manufacturer, to ensure a high genotyping success rate. Eight polymorphisms in the IL1B gene met the criteria and were selected, including the SNPs rs3917368 (A/G at 3′ UTR), rs2853550 (C/T at 3′ UTR), rs1143643 (A/G at intron 6), rs1143634 (C/T at exon 5, known as +3954A/G and +3962A/G), rs1143630 (A/C at intron 3), rs1143627 (C/T at 5′-UTR, known as −31C/T), rs16944 (C/T at 5′-UTR, known as −511C/T), and rs12621220 (C/T at 5′-UTR). 
Genomic DNA Extraction and Genotyping
All blood samples from individuals were collected by venipuncture for genomic DNA isolation. The genomic DNA was extracted from peripheral blood leukocytes (Genomic DNA kit; Qiagen, Valencia, CA) in accordance with the manufacturer's instructions. DNA concentration was quantified in all samples before genotyping. Thirteen of the 484 samples were excluded in this study because the amount of DNA was not enough to perform the assay. All eight single-nucleotide polymorphisms (SNPs) in IL1B were genotyped with an allele-specific extension and ligation assay according to the manufacturer's instructions (Illumina). 
IL1β Quantitative Measurement
The plasma IL1β level was measured by using a quantitative enzyme-linked immunosorbent assay according to the manufacturer's instructions (eBioscience, San Diego, CA). 
Statistical Analysis
Associations between each SNP and disease were assessed by χ2 test. Allele and genotype frequencies in cases and controls were compared and odds rations (ORs) per SNP were estimated by applying unconditional logistic regression. Alleles and genotype frequencies of alleles, genotypes, haplotypes, and diplotypes were expressed as a percentage of the total number of alleles, genotypes, haplotypes, and diplotypes. Results reaching P < 0.05 were statistically significant. The OR, with the 95% confidence interval (CI), was calculated from the genotype and allelic frequencies. Associations between each SNP and IL1β plasma levels were assessed by Student's t-test (for two-category variable) or ANOVA (for a three or more category variable; SPSS for Windows, ver., 14.0; Chicago, IL). Haplotypes were inferred by using Phase 2.1, a computational tool based on Bayesian methods. 31 Linkage disequilibrium (LD) was performed with Haploview 4.1. 31 The multifactor dimensionality reduction (MDR) 1.1.0 of the open-source MDR software package (Dartmouth Medical School, Hanover, NH) was used to detect the best locus–locus interaction models with an estimated testing accuracy of >50% consistency. The interaction dendrogram was established according to a hierarchical clustering algorithm. 32 35  
Results
Basic Characteristics of Patients with GD and Correlation between the Factors
The demographics and clinical information of the participants are summarized in Table 1. We also examined the association of GO with age, sex, smoking status, recurrence, treatment, and clinical features. The χ2 test and Mann-Whitney U test revealed that sex, smoking status, and radioiodine therapy were significantly associated with GO among patients with GD. 
Table 1.
 
Background and Demographic Characteristics of Healthy Individuals and Graves' Patients with or without GO
Table 1.
 
Background and Demographic Characteristics of Healthy Individuals and Graves' Patients with or without GO
Patients' Characteristics Healthy (n = 160) GD/GO (n = 200) GD/non-GO (n = 271) P GD vs. Healthy P GD/GO vs. GD/nonGO
Age at diagnosis (mean ± SD) 27.4 ± 6.4 37.5 ± 10.8 41.7 ± 12.8 1.028 × 10−34 * 2.978 × 10−4 *
Sex
    Male 32 (20.0) 51 (25.5) 48 (17.7) 0.784† 0.040
    Female 128 (80.0) 149 (74.5) 223 (82.3)
Smoking status‡
    Smoking 57 (28.5) 54 (19.9) 0.030
    Nonsmoking 146 (71.5) 217 (80.1)
Recurrence
    Yes 99 (49.5) 128 (47.2) NS†
    No 101 (50.5) 143 (52.8)
Treatment
    Radioiodine
        Yes 15 (7.5) 6 (2.2) 0.006
        No 185 (92.5) 265 (97.8)
    Thyroid gland surgery
        Yes 24 (12.0) 23 (8.5) NS†
        No 176 (88.0) 248 (91.5)
Clinical features
    Goiter
        Grade 1 14 (7.0) 17 (6.3) NS§
        Grade 2 5 (2.5) 21 (7.7)
        Grade 3 22 (11.0) 32 (11.8)
        Grade 4 129 (64.5) 169 (62.4)
        Grade 5 30 (15.0) 32 (11.8)
Nodular hyperplasia
    Yes 22 (11.0) 25 (9.2) NS†
    No 178 (89.0) 246 (90.8)
Myxedema
    Yes 5 (2.5) 1 (0.4) 0.042 *
    No 195 (97.5) 270 (99.6)
Vitiligo
    Yes 2 (1.0) 2 (0.7) NS†
    No 198 (99.0) 269 (99.3)
Allele and Genotype Frequencies of the IL1B Polymorphisms
To identify the SNPs associated with GO, we genotyped eight SNPs in IL1B. As comparing with healthy individuals, the presence of the C allele of SNP rs1143643 may increase the risk of GD (P = 1.655 × 10−112) and GO (P = 1.414 × 10−57), although all eight allele distributions of the IL1B polymorphisms did not differ significantly between GD patients with or without GO (Table 2). Table 3 summarizes the genotype distributions of the IL1B polymorphisms in all individuals. The T/T genotype of SNP rs1143634 was present only in healthy individuals, and the T/T genotype of SNP rs16944 was less frequent in patients with GD (P = 0.041). In addition, the A/A genotype of SNPs rs3917368 and rs1143643 may increase the risk of GO among patients with GD (P = 0.024 and P = 0.017, respectively). The interaction dendrogram of the eight SNPs in the IL1B gene were constructed with the MDR software, and the results revealed a strong interaction of the rs3917368 and rs1143643 loci in the IL1B gene in modulating the risk of GO (Fig. 1). These results suggest that patients with the T/T genotype at SNPs rs1143634 and rs16944 have a lower risk of developing GD. In addition, patients with the A/A genotype of SNPs rs3917368 and rs1143643 may have a higher risk of developing GO. 
Table 2.
 
Allele Frequencies of IL1B Single-Nucleotide Polymorphism in Healthy Individuals and Patients with or without GO
Table 2.
 
Allele Frequencies of IL1B Single-Nucleotide Polymorphism in Healthy Individuals and Patients with or without GO
Alleles Healthy (n = 320) n (%) GD/non-GO (n = 542) n (%) GD/GO (n = 400) n (%) P * (OR, 95% CI)†
GD vs. Healthy GD/GO vs. Healthy GD/GO vs. GD/non-GO
rs3917368
    A allele 171 (53.4) 297 (54.8) 234 (58.5) 0.362 0.174 0.257
    G allele 149 (46.6) 245 (45.2) 166 (41.5)
rs2853550
    C allele 300 (93.8) 505 (93.2) 367 (91.8) 0.478 0.307 0.410
    T allele 20 (6.3) 37 (6.8) 33 (8.3)
rs1143643
    A allele 170 (53.1) 297 (54.8) 235 (58.8) 0.297 0.131 0.226
    G allele 150 (46.9) 245 (45.2) 165 (41.3)
rs1143634
    T allele 168 (52.5) 8 (1.5) 5 (1.3) 1.655 × 10−112 1.414 × 10−57 0.769
    C allele 152 (47.5) 534 (98.5) 395 (98.8) (78.984, 43.795–142.446) (87.316, 35.184–216.689)
rs1143630
    C allele 266 (83.1) 453 (83.6) 332 (83.0) 0.931 0.965 0.814
    A allele 54 (16.9) 89 (16.4) 68 (17.0)
rs1143627
    T allele 182 (56.9) 309 (57.0) 228 (57.0) 0.967 0.973 0.997
    C allele 138 (43.1) 233 (43.0) 172 (43.0)
Rs16944
    C allele 166 (51.9) 309 (57.0) 229 (57.3) 0.090 0.150 0.968
    T allele 154 (48.1) 233 (43.0) 171 (42.8)
rs12621220
    C allele 201 (62.8) 334 (61.6) 256 (64.0) 0.954 0.742 0.456
    T allele 119 (37.2) 208 (38.4) 144 (36.0)
Table 3.
 
Genotype Frequencies of IL1B SNPs in Healthy Individuals and Patients with or without GO
Table 3.
 
Genotype Frequencies of IL1B SNPs in Healthy Individuals and Patients with or without GO
Genotypes Healthy (n = 160) n (%) GD/non-GO (n = 271) n (%) GD/GO (n = 200) n (%) P * OR (95% CI)†
GD vs. Healthy GO vs. Healthy GD/GO vs. GD/non-GO GD vs. Healthy GO vs. Healthy GD/GO vs. GD/non-GO
rs3917368
    A/A 44 (27.5) 70 (25.8) 71 (35.5) 0.594 0.27 0.030 1
    A/G 83 (51.9) 157 (57.9) 92 (46.0) 0.578 (0.380–0.878)
    G/G 33 (20.6) 44 (16.2) 37 (18.5) 0.829 (0.479–1.434)
    A/G+G/G 0.559 0.106 0.024 0.633 (0.425–0.941)
rs2853550
    C/C 141 (88.1) 234 (86.3) 167 (83.5) 0.123 0.202 0.391
    C/T 18 (11.3) 37 (13.7) 33 (16.5)
    T/T 1 (0.6) 0 (0) 0 (0)
    C/T+T/T 0.086 0.263
rs1143643
    A/A 44 (27.5) 70 (25.8) 72 (36.0) 0.495 0.229 0.022 1
    A/G 82 (51.3) 157 (57.9) 91 (45.5) 0.564 (0.371–0.856)
    G/G 34 (21.3) 44 (16.2) 37 (18.5) 0.818 (0.473–1.413)
    A/G+G/G 0.526 0.086 0.017 0.619 (0.416–0.921)
rs1143634
    T/T 53 (33.1) 0 (0) 0 (0) 1 1
    C/T 31 (19.4) 263 (97.0) 195 (97.5) 1.065 × 10−91 2.993 × 10−51 0.767 2.387 × 1010 (0.0–0.0) 10.162 × 109 (0.0–0.0)
    C/C 76 (47.5) 8 (3.0) 5 (2.5) 2.763 × 108 (0.0–0.0) 1.063 × 108 (0.0–0.0)
    C/T+C/C 6.283 × 10−39 1.207 × 10−18 0.767 7.111 × 109 (0.0–0.0) 3.020 × 109 (0.0–0.0)
rs1143630
    C/C 112 (70.0) 186 (68.6) 138 (69.0) 0.447 0.877 0.496
    A/C 42 (26.3) 81 (29.9) 56 (28.0)
    A/A 6 (3.8) 4 (1.5) 6 (3.0)
    A/C+A/A 0.775 0.838 0.933
rs1143627
    T/T 46 (28.8) 83 (30.6) 65 (32.5) 0.528 0.379 0.710
    C/T 90 (56.3) 143 (52.8) 98 (49.0)
    C/C 24 (15.0) 45 (16.6) 37 (18.5)
    C/T+C/C 0.481 0.379 0.592
rs16944
    C/C 48 (29.9) 84 (31.0) 66 (33.0) 0.041 0.21 0.638 1
    C/T 70 (43.8) 143 (52.8) 97 (48.5) 0.911 (0.599–1.387)
    T/T 42 (26.3) 44 (16.2) 37 (18.5) 0.563 (0.356–0.889)
    C/T+T/T 0.664 0.543 0.645
rs12621220
    C/C 58 (36.3) 101 (37.3) 83 (41.5) 0.358 0.296 0.642
    C/T 85 (53.1) 132 (48.7) 90 (45.0)
    T/T 17 (10.6) 38 (14.0) 27 (13.5)
    C/T+T/T 0.527 0.311 0.352
Figure 1.
 
Interaction dendrogram. The location of the longitudinal connecting bars indicates the strength of the dependence: left is weaker and right is stronger. The hierarchical cluster analysis placed IL1B rs3917368 and rs1143643 on the same branch, demonstrating the strong interaction between these two SNPs. There were interactions between IL1B rs3917368-rs1143643 and other SNPs as shown in the dendrogram. IL1B, interleukin-1β gene; SNP, single nucleotide polymorphism.
Figure 1.
 
Interaction dendrogram. The location of the longitudinal connecting bars indicates the strength of the dependence: left is weaker and right is stronger. The hierarchical cluster analysis placed IL1B rs3917368 and rs1143643 on the same branch, demonstrating the strong interaction between these two SNPs. There were interactions between IL1B rs3917368-rs1143643 and other SNPs as shown in the dendrogram. IL1B, interleukin-1β gene; SNP, single nucleotide polymorphism.
Haplotype Frequencies of the IL1B Polymorphisms
Combination of the eight selected SNPs in IL1B by tagging SNPs in the HapMap CHB+JPT population may represent different IL1B haplotypes. In addition, MDR analysis indicated that the best interaction model for predicting the development of GO is the eight-locus model composed by all eight SNPs analyzed in the present study (testing accuracy, 63.9%; cross-validation consistency, 100/100; P < 0.0001). Therefore, we determined the haplotypes in the eight SNPs that had frequencies of >5% and identified the 13 haplotypes shown in Table 4). Ht3-GCGCACTT, Ht5-ACACACTT, and Ht6-GTGCCCTC were found only in patients with GD, whereas Ht9-ACATCTTC, Ht10-ACATCTCC, Ht11-GCGCCCCT, Ht12-ACACCTTC, and HT13-GCGTCCCT were found only in the healthy individuals. Haplotype-specific analysis showed that the Ht1-ACACCTCC and Ht2-GCGCCCTT haplotypes may increase the risk of GD (P = 1.241 × 10−44 and 7.388 × 10−8; OR, 21.599 and 3.917; 95% CI, 12.221–38.175 and 2.304–6.658, respectively) compared with the risk in healthy individuals. Ht4-GCGCCTCC may reduce the risk of GO among the patients with GD (P = 0.025; OR, 0.502; 95% CI, 0.273–0.925). Table 5 showed that 141 patients with GD bore the diplotype A-A/A-A, and it appeared more frequently in patients with GO than did A-A/G-G or G-G/G-G (P = 0.008; OR, 1.650; 95% CI, 1.141–2.384). In 419 patients with GD, the non-Ht4/non-Ht4 diplotype was less frequently found in the patients who also had GO compared with at least one Ht4 haplotype (diplotypes Ht4/Ht4 and Ht4/non-Ht4, P = 0.007; OR, 0.414; 95% CI, 0.214–0.797). These findings confirm that the results from genotype and haplotype analysis. In addition, the LD plots of IL1B in the healthy individuals and GD patients with or without GO showed an apparent variation in these polymorphisms (Fig. 2). These observations suggest that Ht1-ACACCTCC and Ht2-GCGCCCTT haplotypes put the individual at risk for the development of GD, whereas Ht4-GCGCCTCC may play a protective role against the development of GO. 
Table 4.
 
Haplotypes from SNPs of IL1B in Healthy Individuals and Patients with or without GO
Table 4.
 
Haplotypes from SNPs of IL1B in Healthy Individuals and Patients with or without GO
Haplotypes* Healthy n (%) GD P, Global† P, Individual‡ OR (95% CI)§
Non-GO n (%) GO n (%) GD vs. Healthy GD/GO vs. Healthy GD/GO vs. GD/nonGO GD vs. Healthy GD/GO vs. Healthy GD/GO vs. GD/nonGO
Ht1 ACACCTCC 13 (4.1) 251 (46.3) 199 (49.8) 1.241 × 10 −44 9.684 × 10 −41 0.296
21.599 (12.221–38.175) 23.380 (12.979–42.118)
Ht2 GCGCCCTT 16 (5.0) 97 (17.9) 64 (16.0) 7.388 × 10 −8 3.058 × 10 −6 0.445
3.917 (2.304–6.658) 3.619 (2.048–6.395)
Ht3 GCGCACTT 0 (0.0) 53 (9.8) 37 (9.3) 9.598 × 10−9 2.232 × 10−8 0.785
6.068 × 108 (0.0–0.0) 1.424 × 109 (0.0–0.0)
Ht4 GCGCCTCC 13 (4.1) 39 (7.2) 15 (3.8) 0.250 0.829 0.025
0.502 (0.273–0.925)
Ht5 ACACACTT 0 (0.0) 36 (6.6) 31 (7.8) 9.454 × 10−7 3.566 × 10−7 0.513
5.908 × 108 (0.0–0.0) 1.401 × 109 (0.0–0.0)
Ht6 GTGCCCTC 0 (0.0) 14 (2.6) 15 (3.8) 1.496 × 10−3 4.639 × 10−4 0.305
5.662 × 108 (0.0–0.0) 1.343 × 109 (0.0–0.0)
Ht7 GTGCCCTT 1 (0.3) 9 (1.7) 9 (2.3) 1.643 × 10−173 1.793 × 10−100 0.348 0.043 0.273 0.514
6.214 (0.826–46.737)
Ht8 GCGCCCTC 5 (1.6) 12 (2.2) 12 (3.0) 0.310 0.207 0.449
Ht9 ACATCTTC 39 (12.2) 0 (0.0) 0 (0.0) 1.370 × 10−27 7.009 × 10−13
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht10 ACATCTCC 35 (10.9) 0 (0.0) 0 (0.0) 7.485 × 10−25 1.191 × 10−11
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht11 GCGCCCCT 29 (9.1) 0 (0.0) 0 (0.0) 8.971 × 10−21 7.954 × 10−10
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht12 ACACCTTC 23 (7.2) 0 (0.0) 0 (0.0) 1.003 × 10−16 5.047 × 10−8
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht13 GCGTCCCT 23 (7.2) 0 (0.0) 0 (0.0) 1.003 × 10−16 5.047 × 10−8
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Remaining 123 (38.4) 31 (5.7) 18 (4.5)
Total 320 542 400
Table 5.
 
Distribution of IL1B Diplotypes and Their Associations with GO
Table 5.
 
Distribution of IL1B Diplotypes and Their Associations with GO
Diplotypes* Without GO (n = 271) n (%)With GO (n = 200) n (%)Cross Validation ConsistencyPOR (95% CI)
AImage not availableA/AImage not availableA70 (25.8)71 (35.5)1
AImage not availableA/GImage not availableG157 (57.9)92 (44.0)0.030§ 0.578 (0.380–0.878)
GImage not availableG/GImage not availableG44 (14.2)37 (18.5)0.830 (0.480–1.434)‖
AImage not availableA/GImage not availableG + GImage not availableG/GImage not availableG201 (74.2)129 (64.5)0.024§ 0.633 (0.425–0.941)
100/1000.008† 1.650 (1.141–2.384)
Non-Ht4/non-Ht4232 (85.6)187 (93.5)1
Ht4/non-Ht439 (14.4)11 (5.5)0.002§ 0.350 (0.174–0.702)
Ht4/Ht40 (0.0)2 (1.0)2.004 × 109 (0.0–0.0)‖
Ht4/Ht4 + Ht4/non-Ht439 (75.0)13 (25.0)0.007§ 0.414 (0.214–0.797)
Figure 2.
 
Pairwise LD measures of D′ (top) and r 2 (bottom) for the SNPs of the IL1B locus. The scale above the figures indicates the site of each SNP around the IL1B gene region in healthy individuals (A), patients with Graves' disease without ophthalmopathy (B), and patients with Graves' disease with ophthalmopathy (C).
Figure 2.
 
Pairwise LD measures of D′ (top) and r 2 (bottom) for the SNPs of the IL1B locus. The scale above the figures indicates the site of each SNP around the IL1B gene region in healthy individuals (A), patients with Graves' disease without ophthalmopathy (B), and patients with Graves' disease with ophthalmopathy (C).
GO-Associated IL1B Polymorphisms Related to Circulating IL1β Levels among Patients with GD
We performed enzyme-linked immunosorbent assays on plasma proteins from 432 of 471 GD patients and examined their association with IL1B genotypes and diplotypes (Table 6). The mean plasma IL1β concentration in patients with GO was 181.5 ± 580.1 pg/mL, which was significantly higher than in those without GO (88.8 ± 190.9 pg/mL, P = 0.038). Stratified analysis showed that patients with Ht4-GCGCCTCC had much lower concentrations of IL1β (P = 0.042). Although patients with the A-A/G-G and G-G/G-G diplotypes of the SNPs rs3917368 and rs1143643 are less susceptible to GO (GO versus nonGO, 45.9% vs. 57.8%, respectively), they had unexpectedly higher IL1β concentrations than did those with the A-A/A-A diplotype (P = 0.029). Genotype analysis is consistent with the observation. Our results demonstrated that higher circulating IL1β concentrations may correlate with GO development. The GO-protective haplotype Ht4-GCGCCTCC may be used to predict IL1β-induced GO in patients with GD. However, patients with A-A/G-G and G-G/G-G, although less susceptible to GO, may have higher plasma IL1β concentrations than do those with A-A/A-A. These findings imply that the involvement of IL1β in GO development may be more than the IL1B polymorphism-associated circulating IL1β elevation in patients with GD. 
Table 6.
 
Stratified Analysis of Plasma IL1β Levels on GO Risk by IL1B Genotypes
Table 6.
 
Stratified Analysis of Plasma IL1β Levels on GO Risk by IL1B Genotypes
VariablesGD/non-GOGD/GOP * P
88.8 ± 190.9 (249)181.5 ± 580.1 (183)0.038
rs3917368
    A/A62.5 ± 80.9 (63)65.7 ± 124.8 (65)0.868
    A/G102.1 ± 224.8 (144)302.8 ± 825.9 (84)0.0300.032
    G/G81.0 ± 180.1 (42)103.7 ± 200.3 (34)0.606
P = 0.377‡ P = 0.031
    A/G+G/G97.3 ± 215.2 (186)245.4 ± 709.5 (118)0.029
rs1143643
    A/A62.5 ± 80.9 (63)65.8 ± 123.9 (66)0.859
    A/G102.1 ± 224.8 (144)305.5 ± 830.6 (83)0.0290.031
    G/G81.0 ± 180.1 (42)103.7 ± 200.3 (34)0.606
P = 0.377‡ P = 0.029
    A/G+G/G97.3 ± 215.2 (186)246.8 ± 712.4 (117)0.029
    AImage not availableA/AImage not availableA62.5 ± 80.9 (63)65.6 ± 124.8 (65)0.868
    AImage not availableA/GImage not availableG102.1 ± 224.8 (144)302.8 ± 825.9 (84)0.0300.032
    GImage not availableG/GImage not availableG81.0 ± 180.1 (42)103.7 ± 200.3 (34)0.606
P = 0.377‡ P = 0.031
    AImage not availableA/GImage not availableG + GImage not availableG/GImage not availableG97.3 ± 215.2 (186)245.4 ± 709.5 (118)0.029
    Non-Ht4/non-Ht491.1 ± 202.0 (213)189.0 ± 598.1 (171)0.042
    Ht4/non-Ht473.1 ± 103.7 (36)86.6 ± 178.5 (10)0.6040.042
    Ht4/Ht414.2 ± 10.9 (2)0.758
P = 0.402‡ P = 0.547‡
    Ht4/non-Ht4 + Ht4/Ht473.1 ± 103.7 (36)74.6 ± 163.9 (12)0.042
Discussion
We found that the SNPs rs3917368 and rs1143643 in the 3′ UTR and intron regions of IL1B, respectively, may be risk genotypes for development of GO. Persons with the genotypes containing both rs3917368 A/A and rs1143643 A/A may bear a higher risk of developing GO. Clinical association studies showed that the presence of the A-A/A-A diplotype was significantly associated with a higher risk of GO, but the presence of the diplotype along with the Ht4-GCGCCTCC haplotype may be protective against GO. In addition, the circulating IL1β concentrations have been analyzed in GD patients and the association with GO as well as genotypes have been made. Our results showed that IL1B polymorphisms may be associated with the level of circulating IL1β as well as GO development in patients with GD. 
Several reports have predicted the genetic association of IL1B with the development of GD and GO. Hayashi et al. 36 found a significant association between GD and the −31T (rs1143627) allele in the promoter region in their study of the Japanese population. Liu et al. 13 found an association between −511C/T (rs16944) and GD with GO in China, whereas Lacka et al. 14 and Khalilzadeh et al. 15 did not find an association between the IL1B polymorphisms –511C/T (rs16944) and +3954 C/T (rs1143634) and GO in their studies in Iran and Poland, respectively. We found that the C allele of +3954 (rs1143634) is related to the development of GD and GO. In addition, the T/T genotype of −511 (rs16944) is related to less susceptibility to GD. However, in the present study, the genotypes −31C/T (rs1143627) and –511C/T (rs16944) were not associated with the development of GO. The effect of population differences in the determination of such associations should not be underestimated. In the present study, for the first time, we found the association between A/A genotypes of rs3917368 and rs1143643 and the risk for GO, although the significance was weak. Although we identified haplotypes with the Phase program, we found much more diverse haplotype distribution in healthy individuals than in the patients with GD. In addition, patients with GO had more similar haplotype distribution than did those without GO. This observation is consistent with our results from LD analysis. These results may provide new information for prediction of development of GD and GO by different IL1B haplotypes. 
Recent studies have shown that a positive feedback cycle composed of mechanical, immunologic, and cellular processes is involved in the development of GO. 8,21,22 Interstitial accumulation of GAGs, PGE2, and adipogenesis in the orbital tissue contributes to GO in GD patients and these observations and a series of thyroid-related factors, such as the thyrotropin receptor antigen, are thought to be a consequence of the release of certain cytokines, including IL1β, from T cells, monocytes, and activated fibroblasts. 8,21 23,37 Polymorphisms may cause alternations in the expression and function of IL1B, 16 18 which may affect the downregulation of T-cell activation and the subsequent inflammatory diseases, autoimmune diseases, cancer, and GAG accumulation. Given the important role of T cells in the pathogenesis of GO, IL1B may be a candidate gene for the induction of these autoimmune reactions. Our results demonstrated that GO patients have higher plasma IL1β levels than those without GO, which supports this hypothesis. 
Although our hypothesis was that polymorphisms of IL1B relate to the development of GO in patients with GD, the −31C and −511T of IL1B, the polymorphism associated with decreased and increased transcriptional activity, 38,39 were associated with neither IL1β levels nor GO development in our study. We found that the Ht4-GCGCCTCC is related to lower circulating IL1β concentration and less susceptibility toward GO, indicating that Ht4-GCGCCTCC may be used to predict the IL1β-induced GO in patients with GD. However, the A-A/G-G and G-G/G-G diplotypes (less susceptible for GO were related to high circulating levels of IL1β. It is notable that the IL1β concentration examined in our patients (0–3.843 ng/mL) was lower than the IL1β dosage most used in the in vitro experiments (10 ng/mL), 19,20,40,41 and the outliers (IL1β >1 ng/mL) were only the patients with the A-A/G-G genotype. Although plasma IL1β in healthy individuals is yet to be determined, it would be interesting to know whether there are critical dose-dependent levels of IL1β in protection against or promotion of the development of GO. The linkage among the IL1B polymorphisms, IL1β level, and GO development should be further confirmed in studies with larger samples. 
In the present study, our results suggest that IL1B genotypes are associated with the development of GO, the most common orbital disease in GD. This report provides evidence from examination of patient outcomes that polymorphisms of the IL1B gene may predict the development of GO. 
Footnotes
 Supported by Grant CMU96-081 from China Medical University and Grant DMR-95-007 from China Medical University Hospital, Taichung, Taiwan.
Footnotes
 Disclosure: Y.-H. Liu, None; R.-H. Chen, None; H.-H. Wu, None; W.-L. Liao, None; W.-C. Chen, None; Y. Tsai, None; C.-H. Tsai, None; L. Wan, None; F.-J. Tsai, None
The authors thank Yu-Huei Liang and Su-Ching Liu for technical assistance in analyzing the polymorphisms. 
References
Mishra A Mishra SK . Multicentre study of thyroid nodules in patients with Graves' disease (comment on Br J Surg 2000;87:1111–1113). Br J Surg. 2001;88:313. [CrossRef] [PubMed]
Gianoukakis AG Khadavi N Smith TJ . Cytokines, Graves' disease, and thyroid-associated ophthalmopathy. Thyroid. 2008;18:953–958. [CrossRef] [PubMed]
Perros P Neoh C Dickinson J . Thyroid eye disease. BMJ. 2009;338:560. [CrossRef]
Kuriyan AE Phipps RP Feldon SE . The eye and thyroid disease. Curr Opin Ophthalmol. 2008;19:499–506. [CrossRef] [PubMed]
Khoo TK Bahn RS . Pathogenesis of Graves' ophthalmopathy: the role of autoantibodies. Thyroid. 2007;17:1013–1018. [CrossRef] [PubMed]
Kloprogge S Kowal L Wall J Frauman AG . The clinicopathologic basis of Graves' ophthalmopathy: a review. Eur J Ophthalmol. 2005;15:315–323. [PubMed]
Bartalena L . Editorial: glucocorticoids for Graves' ophthalmopathy: how and when. J Clin Endocrinol Metab. 2005;90:5497–5499. [CrossRef] [PubMed]
Bednarczuk T Gopinath B Ploski R Wall JR . Susceptibility genes in Graves' ophthalmopathy: searching for a needle in a haystack? Clin Endocrinol (Oxf). 2007;67:3–19. [CrossRef] [PubMed]
Rasmussen AK Bendtzen K Feldt-Rasmussen U . Thyrocyte-interleukin-1 interactions. Exp Clin Endocrinol Diabetes. 2000;108:67–71. [CrossRef] [PubMed]
Dinarello CA . Biologic basis for interleukin-1 in disease. Blood. 1996;87:2095–2147. [PubMed]
Gianoukakis AG Jennings TA King CS . Hyaluronan accumulation in thyroid tissue: evidence for contributions from epithelial cells and fibroblasts. Endocrinology. 2007;148:54–62. [CrossRef] [PubMed]
Cawood TJ Moriarty P O'Farrelly C O'Shea D . The effects of tumour necrosis factor-alpha and interleukin1 on an in vitro model of thyroid-associated ophthalmopathy: contrasting effects on adipogenesis. Eur J Endocrinol. 2006;155:395–403. [CrossRef] [PubMed]
Liu N Li X Liu C Zhao Y Cui B Ning G . The association of interleukin-1alpha and interleukin-1beta polymorphisms with the risk of Graves' disease in a case-control study and meta-analysis. Hum Immunol. 2010;71:397–401. [CrossRef] [PubMed]
Lacka K Paradowska A Gasinska T . Interleukin-1beta gene (IL-1beta) polymorphisms (SNP -511 and SNP +3953) in thyroid-associated ophthalmopathy (TAO) among the Polish population. Curr Eye Res. 2009;34:215–220. [CrossRef] [PubMed]
Khalilzadeh O Mojazi Amiri H Tahvildari M . Pretibial myxedema is associated with polymorphism in exon 1 of CTLA-4 gene in patients with Graves' ophthalmopathy. Arch Dermatol Res. 2009;301:719–723. [CrossRef] [PubMed]
Qian N Chen X Han S . Circulating IL-1beta levels, polymorphisms of IL-1B, and risk of cervical cancer in Chinese women. J Cancer Res Clin Oncol. 2010;136:709–716. [CrossRef] [PubMed]
Garcia-Gonzalez MA Aisa MA Strunk M . Relevance of IL-1 and TNF gene polymorphisms on interleukin-1beta and tumor necrosis factor-alpha gastric mucosal production. Hum Immunol. 2009;70:935–945. [CrossRef] [PubMed]
Landvik NE Hart K Skaug V Stangeland LB Haugen A Zienolddiny S . A specific interleukin-1B haplotype correlates with high levels of IL1B mRNA in the lung and increased risk of non-small cell lung cancer. Carcinogenesis. 2009;30:1186–1192. [CrossRef] [PubMed]
Wong YK Tang KT Wu JC Hwang JJ Wang HS . Stimulation of hyaluronan synthesis by interleukin-1beta involves activation of protein kinase C betaII in fibroblasts from patients with Graves' ophthalmopathy. J Cell Biochem. 2001;82:58–67. [CrossRef] [PubMed]
Hwang CJ Afifiyan N Sand D . Orbital fibroblasts from patients with thyroid-associated ophthalmopathy overexpress CD40: CD154 hyperinduces IL-6, IL-8, and MCP-1. Invest Ophthalmol Vis Sci. 2009;50:2262–2268. [CrossRef] [PubMed]
Prabhakar BS Bahn RS Smith TJ . Current perspective on the pathogenesis of Graves' disease and ophthalmopathy. Endocr Rev. 2003;24:802–835. [CrossRef] [PubMed]
Bahn RS . Pathophysiology of Graves' ophthalmopathy: the cycle of disease. Clinical review 157. J Clin Endocrinol Metab. 2003;88:1939–1946. [CrossRef] [PubMed]
Heufelder AE . Pathogenesis of ophthalmopathy in autoimmune thyroid disease. Rev Endocr Metab Disord. 2000;1:87–95. [CrossRef] [PubMed]
Tsai LJ Hsiao SH Tsai JJ Lin CY Tsai LM Lan JL . Higher genetic susceptibility to inflammation in mild disease activity of systemic lupus erythematosus. Rheumatol Int. 2009;29:1001–1011. [CrossRef] [PubMed]
Parks CG Cooper GS Dooley MA . Systemic lupus erythematosus and genetic variation in the interleukin 1 gene cluster: a population based study in the southeastern United States. Ann Rheum Dis. 2004;63:91–94. [CrossRef] [PubMed]
Arnett FC . Genetic studies of human lupus in families. Int Rev Immunol. 2000;19:297–317. [CrossRef] [PubMed]
Zini N Lisignoli G Solimando L . IL1-beta and TNF-alpha induce changes in the nuclear polyphosphoinositide signalling system in osteoblasts similar to that occurring in patients with rheumatoid arthritis: an immunochemical and immunocytochemical study. Histochem Cell Biol. 2003;120:243–250. [CrossRef] [PubMed]
Johnsen AK Plenge RM Butty V . A broad analysis of IL1 polymorphism and rheumatoid arthritis. Arthritis Rheum. 2008;58:1947–1957. [CrossRef] [PubMed]
Fagiolo E Vigevani F Pozzetto U . High cytokine serum levels in patients with autoimmune hemolytic anemia (AIHA). Immunol Invest. 1994;23:449–456. [CrossRef] [PubMed]
Chen RH Chen WC Chang CT Tsai CH Tsai FJ . Interleukin-1-beta gene, but not the interleukin-1 receptor antagonist gene, is associated with Graves' disease. J Clin Lab Anal. 2005;19:133–138. [CrossRef] [PubMed]
Barrett JC Fry B Maller J Daly MJ . Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics. 2005;21:263–265. [CrossRef] [PubMed]
Ritchie MD Hahn LW Roodi N . Multifactor-dimensionality reduction reveals high-order interactions among estrogen-metabolism genes in sporadic breast cancer. Am J Hum Genet. 2001;69:138–147. [CrossRef] [PubMed]
Hahn LW Ritchie MD Moore JH . Multifactor dimensionality reduction software for detecting gene-gene and gene-environment interactions. Bioinformatics. 2003;19:376–382. [CrossRef] [PubMed]
Coffey CS Hebert PR Ritchie MD . An application of conditional logistic regression and multifactor dimensionality reduction for detecting gene-gene interactions on risk of myocardial infarction: the importance of model validation. BMC Bioinformatics. 2004;5:49. [CrossRef] [PubMed]
Ritchie MD Hahn LW Moore JH . Power of multifactor dimensionality reduction for detecting gene-gene interactions in the presence of genotyping error, missing data, phenocopy, and genetic heterogeneity. Genet Epidemiol. 2003;24:150–157. [CrossRef] [PubMed]
Hayashi F Watanabe M Nanba T Inoue N Akamizu T Iwatani Y . Association of the −31C/T functional polymorphism in the interleukin-1beta gene with the intractability of Graves' disease and the proportion of T helper type 17 cells. Clin Exp Immunol. 2009;158:281–289. [CrossRef] [PubMed]
Fatourechi V . Pretibial myxedema: pathophysiology and treatment options. Am J Clin Dermatol. 2005;6:295–309. [CrossRef] [PubMed]
Lind H Haugen A Zienolddiny S . Differential binding of proteins to the IL1B −31 T/C polymorphism in lung epithelial cells. Cytokine. 2007;38:43–48. [CrossRef] [PubMed]
Chen H Wilkins LM Aziz N . Single nucleotide polymorphisms in the human interleukin-1B gene affect transcription according to haplotype context. Hum Mol Genet. 2006;15:519–529. [CrossRef] [PubMed]
Han R Smith TJ . T helper type 1 and type 2 cytokines exert divergent influence on the induction of prostaglandin E2 and hyaluronan synthesis by interleukin-1beta in orbital fibroblasts: implications for the pathogenesis of thyroid-associated ophthalmopathy. Endocrinology. 2006;147:13–19. [CrossRef] [PubMed]
Chen B Tsui S Smith TJ . IL-1 beta induces IL-6 expression in human orbital fibroblasts: identification of an anatomic-site specific phenotypic attribute relevant to thyroid-associated ophthalmopathy. J Immunol. 2005;175:1310–1319. [CrossRef] [PubMed]
Figure 1.
 
Interaction dendrogram. The location of the longitudinal connecting bars indicates the strength of the dependence: left is weaker and right is stronger. The hierarchical cluster analysis placed IL1B rs3917368 and rs1143643 on the same branch, demonstrating the strong interaction between these two SNPs. There were interactions between IL1B rs3917368-rs1143643 and other SNPs as shown in the dendrogram. IL1B, interleukin-1β gene; SNP, single nucleotide polymorphism.
Figure 1.
 
Interaction dendrogram. The location of the longitudinal connecting bars indicates the strength of the dependence: left is weaker and right is stronger. The hierarchical cluster analysis placed IL1B rs3917368 and rs1143643 on the same branch, demonstrating the strong interaction between these two SNPs. There were interactions between IL1B rs3917368-rs1143643 and other SNPs as shown in the dendrogram. IL1B, interleukin-1β gene; SNP, single nucleotide polymorphism.
Figure 2.
 
Pairwise LD measures of D′ (top) and r 2 (bottom) for the SNPs of the IL1B locus. The scale above the figures indicates the site of each SNP around the IL1B gene region in healthy individuals (A), patients with Graves' disease without ophthalmopathy (B), and patients with Graves' disease with ophthalmopathy (C).
Figure 2.
 
Pairwise LD measures of D′ (top) and r 2 (bottom) for the SNPs of the IL1B locus. The scale above the figures indicates the site of each SNP around the IL1B gene region in healthy individuals (A), patients with Graves' disease without ophthalmopathy (B), and patients with Graves' disease with ophthalmopathy (C).
Table 1.
 
Background and Demographic Characteristics of Healthy Individuals and Graves' Patients with or without GO
Table 1.
 
Background and Demographic Characteristics of Healthy Individuals and Graves' Patients with or without GO
Patients' Characteristics Healthy (n = 160) GD/GO (n = 200) GD/non-GO (n = 271) P GD vs. Healthy P GD/GO vs. GD/nonGO
Age at diagnosis (mean ± SD) 27.4 ± 6.4 37.5 ± 10.8 41.7 ± 12.8 1.028 × 10−34 * 2.978 × 10−4 *
Sex
    Male 32 (20.0) 51 (25.5) 48 (17.7) 0.784† 0.040
    Female 128 (80.0) 149 (74.5) 223 (82.3)
Smoking status‡
    Smoking 57 (28.5) 54 (19.9) 0.030
    Nonsmoking 146 (71.5) 217 (80.1)
Recurrence
    Yes 99 (49.5) 128 (47.2) NS†
    No 101 (50.5) 143 (52.8)
Treatment
    Radioiodine
        Yes 15 (7.5) 6 (2.2) 0.006
        No 185 (92.5) 265 (97.8)
    Thyroid gland surgery
        Yes 24 (12.0) 23 (8.5) NS†
        No 176 (88.0) 248 (91.5)
Clinical features
    Goiter
        Grade 1 14 (7.0) 17 (6.3) NS§
        Grade 2 5 (2.5) 21 (7.7)
        Grade 3 22 (11.0) 32 (11.8)
        Grade 4 129 (64.5) 169 (62.4)
        Grade 5 30 (15.0) 32 (11.8)
Nodular hyperplasia
    Yes 22 (11.0) 25 (9.2) NS†
    No 178 (89.0) 246 (90.8)
Myxedema
    Yes 5 (2.5) 1 (0.4) 0.042 *
    No 195 (97.5) 270 (99.6)
Vitiligo
    Yes 2 (1.0) 2 (0.7) NS†
    No 198 (99.0) 269 (99.3)
Table 2.
 
Allele Frequencies of IL1B Single-Nucleotide Polymorphism in Healthy Individuals and Patients with or without GO
Table 2.
 
Allele Frequencies of IL1B Single-Nucleotide Polymorphism in Healthy Individuals and Patients with or without GO
Alleles Healthy (n = 320) n (%) GD/non-GO (n = 542) n (%) GD/GO (n = 400) n (%) P * (OR, 95% CI)†
GD vs. Healthy GD/GO vs. Healthy GD/GO vs. GD/non-GO
rs3917368
    A allele 171 (53.4) 297 (54.8) 234 (58.5) 0.362 0.174 0.257
    G allele 149 (46.6) 245 (45.2) 166 (41.5)
rs2853550
    C allele 300 (93.8) 505 (93.2) 367 (91.8) 0.478 0.307 0.410
    T allele 20 (6.3) 37 (6.8) 33 (8.3)
rs1143643
    A allele 170 (53.1) 297 (54.8) 235 (58.8) 0.297 0.131 0.226
    G allele 150 (46.9) 245 (45.2) 165 (41.3)
rs1143634
    T allele 168 (52.5) 8 (1.5) 5 (1.3) 1.655 × 10−112 1.414 × 10−57 0.769
    C allele 152 (47.5) 534 (98.5) 395 (98.8) (78.984, 43.795–142.446) (87.316, 35.184–216.689)
rs1143630
    C allele 266 (83.1) 453 (83.6) 332 (83.0) 0.931 0.965 0.814
    A allele 54 (16.9) 89 (16.4) 68 (17.0)
rs1143627
    T allele 182 (56.9) 309 (57.0) 228 (57.0) 0.967 0.973 0.997
    C allele 138 (43.1) 233 (43.0) 172 (43.0)
Rs16944
    C allele 166 (51.9) 309 (57.0) 229 (57.3) 0.090 0.150 0.968
    T allele 154 (48.1) 233 (43.0) 171 (42.8)
rs12621220
    C allele 201 (62.8) 334 (61.6) 256 (64.0) 0.954 0.742 0.456
    T allele 119 (37.2) 208 (38.4) 144 (36.0)
Table 3.
 
Genotype Frequencies of IL1B SNPs in Healthy Individuals and Patients with or without GO
Table 3.
 
Genotype Frequencies of IL1B SNPs in Healthy Individuals and Patients with or without GO
Genotypes Healthy (n = 160) n (%) GD/non-GO (n = 271) n (%) GD/GO (n = 200) n (%) P * OR (95% CI)†
GD vs. Healthy GO vs. Healthy GD/GO vs. GD/non-GO GD vs. Healthy GO vs. Healthy GD/GO vs. GD/non-GO
rs3917368
    A/A 44 (27.5) 70 (25.8) 71 (35.5) 0.594 0.27 0.030 1
    A/G 83 (51.9) 157 (57.9) 92 (46.0) 0.578 (0.380–0.878)
    G/G 33 (20.6) 44 (16.2) 37 (18.5) 0.829 (0.479–1.434)
    A/G+G/G 0.559 0.106 0.024 0.633 (0.425–0.941)
rs2853550
    C/C 141 (88.1) 234 (86.3) 167 (83.5) 0.123 0.202 0.391
    C/T 18 (11.3) 37 (13.7) 33 (16.5)
    T/T 1 (0.6) 0 (0) 0 (0)
    C/T+T/T 0.086 0.263
rs1143643
    A/A 44 (27.5) 70 (25.8) 72 (36.0) 0.495 0.229 0.022 1
    A/G 82 (51.3) 157 (57.9) 91 (45.5) 0.564 (0.371–0.856)
    G/G 34 (21.3) 44 (16.2) 37 (18.5) 0.818 (0.473–1.413)
    A/G+G/G 0.526 0.086 0.017 0.619 (0.416–0.921)
rs1143634
    T/T 53 (33.1) 0 (0) 0 (0) 1 1
    C/T 31 (19.4) 263 (97.0) 195 (97.5) 1.065 × 10−91 2.993 × 10−51 0.767 2.387 × 1010 (0.0–0.0) 10.162 × 109 (0.0–0.0)
    C/C 76 (47.5) 8 (3.0) 5 (2.5) 2.763 × 108 (0.0–0.0) 1.063 × 108 (0.0–0.0)
    C/T+C/C 6.283 × 10−39 1.207 × 10−18 0.767 7.111 × 109 (0.0–0.0) 3.020 × 109 (0.0–0.0)
rs1143630
    C/C 112 (70.0) 186 (68.6) 138 (69.0) 0.447 0.877 0.496
    A/C 42 (26.3) 81 (29.9) 56 (28.0)
    A/A 6 (3.8) 4 (1.5) 6 (3.0)
    A/C+A/A 0.775 0.838 0.933
rs1143627
    T/T 46 (28.8) 83 (30.6) 65 (32.5) 0.528 0.379 0.710
    C/T 90 (56.3) 143 (52.8) 98 (49.0)
    C/C 24 (15.0) 45 (16.6) 37 (18.5)
    C/T+C/C 0.481 0.379 0.592
rs16944
    C/C 48 (29.9) 84 (31.0) 66 (33.0) 0.041 0.21 0.638 1
    C/T 70 (43.8) 143 (52.8) 97 (48.5) 0.911 (0.599–1.387)
    T/T 42 (26.3) 44 (16.2) 37 (18.5) 0.563 (0.356–0.889)
    C/T+T/T 0.664 0.543 0.645
rs12621220
    C/C 58 (36.3) 101 (37.3) 83 (41.5) 0.358 0.296 0.642
    C/T 85 (53.1) 132 (48.7) 90 (45.0)
    T/T 17 (10.6) 38 (14.0) 27 (13.5)
    C/T+T/T 0.527 0.311 0.352
Table 4.
 
Haplotypes from SNPs of IL1B in Healthy Individuals and Patients with or without GO
Table 4.
 
Haplotypes from SNPs of IL1B in Healthy Individuals and Patients with or without GO
Haplotypes* Healthy n (%) GD P, Global† P, Individual‡ OR (95% CI)§
Non-GO n (%) GO n (%) GD vs. Healthy GD/GO vs. Healthy GD/GO vs. GD/nonGO GD vs. Healthy GD/GO vs. Healthy GD/GO vs. GD/nonGO
Ht1 ACACCTCC 13 (4.1) 251 (46.3) 199 (49.8) 1.241 × 10 −44 9.684 × 10 −41 0.296
21.599 (12.221–38.175) 23.380 (12.979–42.118)
Ht2 GCGCCCTT 16 (5.0) 97 (17.9) 64 (16.0) 7.388 × 10 −8 3.058 × 10 −6 0.445
3.917 (2.304–6.658) 3.619 (2.048–6.395)
Ht3 GCGCACTT 0 (0.0) 53 (9.8) 37 (9.3) 9.598 × 10−9 2.232 × 10−8 0.785
6.068 × 108 (0.0–0.0) 1.424 × 109 (0.0–0.0)
Ht4 GCGCCTCC 13 (4.1) 39 (7.2) 15 (3.8) 0.250 0.829 0.025
0.502 (0.273–0.925)
Ht5 ACACACTT 0 (0.0) 36 (6.6) 31 (7.8) 9.454 × 10−7 3.566 × 10−7 0.513
5.908 × 108 (0.0–0.0) 1.401 × 109 (0.0–0.0)
Ht6 GTGCCCTC 0 (0.0) 14 (2.6) 15 (3.8) 1.496 × 10−3 4.639 × 10−4 0.305
5.662 × 108 (0.0–0.0) 1.343 × 109 (0.0–0.0)
Ht7 GTGCCCTT 1 (0.3) 9 (1.7) 9 (2.3) 1.643 × 10−173 1.793 × 10−100 0.348 0.043 0.273 0.514
6.214 (0.826–46.737)
Ht8 GCGCCCTC 5 (1.6) 12 (2.2) 12 (3.0) 0.310 0.207 0.449
Ht9 ACATCTTC 39 (12.2) 0 (0.0) 0 (0.0) 1.370 × 10−27 7.009 × 10−13
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht10 ACATCTCC 35 (10.9) 0 (0.0) 0 (0.0) 7.485 × 10−25 1.191 × 10−11
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht11 GCGCCCCT 29 (9.1) 0 (0.0) 0 (0.0) 8.971 × 10−21 7.954 × 10−10
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht12 ACACCTTC 23 (7.2) 0 (0.0) 0 (0.0) 1.003 × 10−16 5.047 × 10−8
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Ht13 GCGTCCCT 23 (7.2) 0 (0.0) 0 (0.0) 1.003 × 10−16 5.047 × 10−8
0.000 (0.0–0.0) 0.000 (0.0–0.0)
Remaining 123 (38.4) 31 (5.7) 18 (4.5)
Total 320 542 400
Table 5.
 
Distribution of IL1B Diplotypes and Their Associations with GO
Table 5.
 
Distribution of IL1B Diplotypes and Their Associations with GO
Diplotypes* Without GO (n = 271) n (%)With GO (n = 200) n (%)Cross Validation ConsistencyPOR (95% CI)
AImage not availableA/AImage not availableA70 (25.8)71 (35.5)1
AImage not availableA/GImage not availableG157 (57.9)92 (44.0)0.030§ 0.578 (0.380–0.878)
GImage not availableG/GImage not availableG44 (14.2)37 (18.5)0.830 (0.480–1.434)‖
AImage not availableA/GImage not availableG + GImage not availableG/GImage not availableG201 (74.2)129 (64.5)0.024§ 0.633 (0.425–0.941)
100/1000.008† 1.650 (1.141–2.384)
Non-Ht4/non-Ht4232 (85.6)187 (93.5)1
Ht4/non-Ht439 (14.4)11 (5.5)0.002§ 0.350 (0.174–0.702)
Ht4/Ht40 (0.0)2 (1.0)2.004 × 109 (0.0–0.0)‖
Ht4/Ht4 + Ht4/non-Ht439 (75.0)13 (25.0)0.007§ 0.414 (0.214–0.797)
Table 6.
 
Stratified Analysis of Plasma IL1β Levels on GO Risk by IL1B Genotypes
Table 6.
 
Stratified Analysis of Plasma IL1β Levels on GO Risk by IL1B Genotypes
VariablesGD/non-GOGD/GOP * P
88.8 ± 190.9 (249)181.5 ± 580.1 (183)0.038
rs3917368
    A/A62.5 ± 80.9 (63)65.7 ± 124.8 (65)0.868
    A/G102.1 ± 224.8 (144)302.8 ± 825.9 (84)0.0300.032
    G/G81.0 ± 180.1 (42)103.7 ± 200.3 (34)0.606
P = 0.377‡ P = 0.031
    A/G+G/G97.3 ± 215.2 (186)245.4 ± 709.5 (118)0.029
rs1143643
    A/A62.5 ± 80.9 (63)65.8 ± 123.9 (66)0.859
    A/G102.1 ± 224.8 (144)305.5 ± 830.6 (83)0.0290.031
    G/G81.0 ± 180.1 (42)103.7 ± 200.3 (34)0.606
P = 0.377‡ P = 0.029
    A/G+G/G97.3 ± 215.2 (186)246.8 ± 712.4 (117)0.029
    AImage not availableA/AImage not availableA62.5 ± 80.9 (63)65.6 ± 124.8 (65)0.868
    AImage not availableA/GImage not availableG102.1 ± 224.8 (144)302.8 ± 825.9 (84)0.0300.032
    GImage not availableG/GImage not availableG81.0 ± 180.1 (42)103.7 ± 200.3 (34)0.606
P = 0.377‡ P = 0.031
    AImage not availableA/GImage not availableG + GImage not availableG/GImage not availableG97.3 ± 215.2 (186)245.4 ± 709.5 (118)0.029
    Non-Ht4/non-Ht491.1 ± 202.0 (213)189.0 ± 598.1 (171)0.042
    Ht4/non-Ht473.1 ± 103.7 (36)86.6 ± 178.5 (10)0.6040.042
    Ht4/Ht414.2 ± 10.9 (2)0.758
P = 0.402‡ P = 0.547‡
    Ht4/non-Ht4 + Ht4/Ht473.1 ± 103.7 (36)74.6 ± 163.9 (12)0.042
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×