Expression of non-thyroid specific autoantibodies among autoimmune thyroiditis patients in Iraq
Saffar3
- , Montaha A.Al Ani2
- Issam AL
1, Jibory -
Ali Majid Al
1,2Middle Technical university-College of Health and Medical Technologies-Department of Medical Laboratory Technology-Baghdad-Iraq.
3 Middle Technical university, Department of community health, Institute of Medical Technology, Baghdad, Iraq.
*Corresponding author E-mail: [email protected]
Abstract
Introduction: Autoimmune thyroid disease (AITD) are the most common organ specific autoimmune disorders affecting 5-10% of populations. AITD is characterized by the presence of autoantibodies to thyroid gland component as anti-Tg and anti-TPO, also the presence of autoantibodies of other diseases as dsDNA. Aim of study: Our study aimed to determine the correlation of AMA and dsDNA autoantibodies with Iraqi AITD patients, also to detect the hormonal changes among patients groups. Materials and methods: Ninety (90) blood samples were collected from patients suffering thyroid gland disease in both gender whose age ranged 15-65 years, for the period from September to the end of November 2020 from Baghdad teaching hospitals, samples divided into 60 patients groups and 30 healthy control. All samples were tested to evaluate the levels of TSH and T3 hormones, AMA and dsDNA autoantibodies by using the ELISA technique. The data obtained were subjected to analysis by using statistical package of social science (SPSS) version 26.0. Results: the highest rate was noticed in the autoimmune hyperthyroidism group (56.81%) to (43.18%) in hypothyroidism group. The percentage rate of incidence was in (15-44) years more than (45-65) years in both autoimmune hyperthyroidism and autoimmune hypothyroidism.
Females represent about 77.27% of studied groups, also our result indicate that non-organ specific autoantibodies (AMA and anti-ds DNA) levels are elevated in patient with autoimmune thyroiditis compared with healthy control group. Conclusion: From the revealed data we conclude that patients with AITD are prone for other autoimmune disease.
Keywords: autoimmune thyroiditis, nonorgan specific autoantibodies, AMA, dsDNA, T3, TSH, non-specific thyroid autoantibodies.
Introduction
The thyroid is an endocrine gland. Its location is in the inferior, anterior neck, and it is responsible for the formation and secretion of the thyroid hormones as well as iodine homeostasis within the human body. The thyroid produces approximately 90% inactive thyroid hormone, or thyroxine (T4), and 10%
active thyroid hormone, or triiodothyronine (T3) (1). One of the main disorders that occur in thyroid gland is autoimmune thyroiditis (AIT) which is a typical organ-specific autoimmune disorder, is the main cause of hypothyroidism and hyperthyroidism affecting 10% of the population. AIT is characterized by the presence of thyroid-specific autoantibodies (anti-thyroid peroxidase (TPOAbs), anti- thyroglobulin (TgAbs) or anti-TSH receptor (TSHRAbs)) in the serum, massive infiltration of lymphocytic cells, and destruction of the follicular structure within the thyroid (2). Genetically autoimmune thyroid diseases (AITD) are characterized through increased familial clustering, reduced DNASE1 gene expression, CTLA-4 polymorphisms, and are strongly associated with the major histocompatibility (MHC) complex (3). Most common types of AIT are GD and HT in which Hashimoto thyroiditis (HT) is the most common clinical presentations of TAI, characterized by the presence of thyroid autoantibodies, including thyroid peroxidase antibodies (TPOAbs) and thyroglobulin antibodies (TgAbs), mediating antibody-dependent cell-mediated cytotoxicity (4). GD is caused by TSHR-stimulating antibodies (TSAb), which act as agonists by stimulating thyroid growth and thyroid hormone synthesis in an unregulated manner. In contrast, blocking anti-TSHR-Ab (TBAb) acts as TSHR antagonists, which block the action of the thyroid-stimulating hormone (TSH) and can cause the hypothyroidism of HT (5). In individuals with anti-dsDNA antibodies and impaired clearance of cell debris including necrotic chromatin, this may change the situation from a controlled removal of chromatin into a condition where chromatin debris remains exposed and may be a contributor to produce and or amplify anti-dsDNA antibodies by interaction with TLR9 and to promote
inflammation (6). Existence of ATD among patients with systemic autoimmune diseases, such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), or Sjögren syndrome (SS), had been well recognized. On the other hand, a high prevalence of other autoantibodies directed against specific non-thyroid antigens has been described in patients with AITD, including anti-double stranded deoxyribonucleic acid (anti-dsDNA) whose clinical meaning is unknown (7). Anti-mitochondrial (AMA) antibodies are identified in approximately 95% of individuals with primary biliary cirrhosis, As well as found in patients with hyperthyroiditis patients. Antibodies against the mitochondrial ATP/ADP transporter protein can be produced by people with autoimmune thyroid illness. Anti- mitochondrial antibodies have been found in patients with hyperthyroiditis, and primary biliary cirrhosis has been linked to autoimmune thyroid illness (8).
Objectives of the study
Our study focused on the following: Determining the correlation of the non-thyroid specific autoantibodies with autoimmune thyroid diseases (AITD), identification of the hormonal changes in autoimmune thyroiditis Iraqi patients, and demonstrate the significant degree of the anti-mitochondrial antibody (AMA) and anti-double stranded DNA (dsDNA) in autoimmune thyroiditis patients.
Materials and methods
The current included a collection of ninety (90) blood samples divided into sixty patients groups whose suffering from thyroid gland diseases and thirty control group in both gender whose aged 15-65 years for the period from September.to the end of November 2020 from educational Laboratories / Baghdad teaching hospital. Serum centrifuged at 3000 xg for 30 min and stored at (-20 to -80 °C) until analysis.
During the sample collection, a questionnaire has been collected by patients that include: Sex, age, family and relative history, smoking, radiation exposure, sera of all 90 sample were analyzed by the ELISA technique to detect the concentrations of the immunological markers AMA ELISA kit, according to manufacturer’s instructions (MyBiosource Co. USA) and dsDNA ELISA Kit, according to manufacturer’s instructions (Cusabio Biotech Co. China). For the biochemical markers (TSH ELISA Kit, Cusabio biotech Co. China and T3 ELISA Kit, Co. USA hormones).
The tests were performed using ELISA, Human Co. LTD. Germany Statistical analysis:
The data obtained were subjected to analysis by using statistical package of social science (SPSS) version 26.0. Results were expressed as mean standard error and values of (P>0.05) were considered statically non-significant. While (P<0.05) and (P<0.01, 0.001), were considered significantly different and highly significantly different respectively.
Result and discussion
In this study, patients have been classified into two groups autoimmune hyperthyroidism and autoimmune hypothyroidism at 25(56.8%) and 19(43.2%) respectively, as shown in Table (1). For the current work it is sufficient to put out that the percentage rate of incidence was in age range (15-44) years more than age range (45-65) years in both autoimmune hyperthyroidism and autoimmune hypothyroidism at 19(76%) and 12(63.16%) respectively.
As noted in our results in Table (1) by analyzing according gender of distribution, the result of studied groups according to gender pointed the majority of autoimmune hyperthyroidism were females at 19(76%) while the frequency of males at 6(24%) and that mean that the females represent about 77.27% of the studied group. There were highly significant differences among the patient’s ages and sex of these groups were (p≤0.01).
The hormone levels of T3 and TSH were utilized to assess if the patients had hyperthyroidism or hypothyroidism. In this study, patients were classified into two group autoimmune hyperthyroidism and autoimmune hypothyroidism, the highest rate was noticed in the autoimmune hyperthyroiditis group (56.81%) to (43.18%) in hypothyroiditis group Table (1). Similar result was obtained by (9) when they found that out of (383) autoimmune thyroiditis patients 94.5% were hyperthyroiditis and 5.5% hypothyroiditis also mentioned that the risk of autoimmune thyroiditis increases with patient’s age.
For the current work it is sufficient to put out that the percentage rate of incidence was in (15-44) years more than (45-65) years in both autoimmune hyperthyroidism and autoimmune hypothyroidism.
As noted in our results by analyzing gender of distribution, the result of studied groups according to gender pointed the majority of autoimmune hyperthyroidism were females at 19 (76%) while the frequency of males at 6(24%) as noted from this distribution females represent about 77.27% of studied group Table (1), several surveys suggested a females are more susceptible to autoimmune disease (10), while (11) has been detected that autoimmune thyroiditis is thought to be a disease that can appear in childhood but is more prevalent during adulthood. And the result are consistent with finding done in Mosul province (12) which has been reported that the peak of age was 30 ± 5.2 years.
The age range of both patients groups in our study was similar to that reported by (13), which indicated that highest percentage of participants (81%) at age group15-44 year and result revealed female gender has categorically significantly high (P≤0.01) percentage of occurrence of thyroid abnormality as compared to male gender 75.8% females vs. 24.2% males. The finding observed in the study are in agreement with Fairweather and his coworkers who reported that numerous theories proposed for clarification of the high incidence in autoimmune diseases in women due to that females are known to interact with trauma, vaccination, and infection with exaggerated antibody secretion and a predominance T helper (Th2) immune reaction, However Th1 response and inflammation are often more extreme in males. Females have comparable number of lymphocytes but higher potential for antibody generation by B cells. As well as, females have much more vigorous humoral and cellular immune reactions and greater than normal CD4+ T cell level following immunization (14).
Table 1 Distribution of patient groups (n=44) between (Autoimmune hyperthyroidism and Autoimmune hypothyroidism) according age group and sex group.
Sex Age(Years)
Patients group
Female Male
No.
(45-65) (15-44)
No.
19 (76.00%) 6
(24.00%) 25
(56.81%) 6
(24.00%) 19
(76.00%) 25
(56.81%) Autoimmune
hyperthyroidism
15 (78.95%) 4
(21.05%) 19
(43.18%) 7 (36.84%)
12 (63.16%) 19
(43.18%) Autoimmune
hypothyroidism
34 (77.27%) 10
(22.73%) 44
13 (29.55%) 31
(70.45%) 44
Total
0.0084 **
-- 0.0037 **
-- P-value
** (P≤0.01)-Highly Significant.
In the present work, patients with autoimmune thyroiditis were studied in the search for potential immunological markers for diagnosis assessment.
Sera of all study groups were estimated for serum AMA and anti-ds DNA, the results from Table (2) revealed the correlation between immunological markers and patients groups.
Our results indicate that non-organ specific autoantibodies (AMA and anti-ds DNA) levels are elevated in patient with autoimmune thyroiditis compared with healthy control group and the major Mean value of non-organ specific autoantibodies was AMA (6.15±3.25) monitored in studied group, while the less Mean value of immunological markers in anti-dsDNA were ( 0.98±0.63 ), Table (2), the proper explanation for the above result is that the patients included in the study are either prone for poly autoimmunity or they have other autoimmune disorder(s) but still asymptomatic. However the patients in the study wasn’t have any features of cirrhosis, SLE or arthritis.
Study by (8) conducted in 1994, reported that thyroid antibodies are present in more than 15% of patients with primary biliary cirrhosis. Inversely, the prevalence of antimitochondrial antibodies in patients with thyroid antibodies is still unknown.
Our result agrees with Iraqi study conducted in Mosel by (15), they reported that from a group of four patients with Hashimoto thyroiditis assessed for AMA a single case was positive. However, the patient was not clinically known to have any features of cirrhosis.
Further study elevate the relation between autoimmune thyroiditis and other autoimmune diseases, the researcher (16) pointed out to the prevalence of autoimmune thyroid disease in patients with autoimmune liver disease (AILD) has been observed. Data on the clinical relationship between AILD and AITD remain scanty. The high incidence of AILD concomitant with AITD, as well as the higher levels of serum IgG and γ-globulin, and the strong correlation between thyroid antibody and IgG, suggesting that we should strengthen the screening of autoimmune thyroid disease when diagnosing and treating autoimmune
On the other hand, the findings observed in studies done by (17), (18) which also have fund elevated levels of anti-dsDNA in AITD patients at 74.5% .Also , a study by (19) conducted evaluate the risk of developing RA in non-Rheumatoid Arthritis AITD patient that found a significant difference from control in the level of anti- ATD is commonly associated with nonorgan-specific autoantibodies.
Increased prevalence of anti-dsDNA autoantibodies in AITDs without any evidence of any rheumatic disease is unclear (20).
In AITDs, lymphocytic infiltration in the thyroid shows up a deranged immune system charging the follicular cells of the thyroid gland (thyrocytes) leading to the condition of either hyperthyroidism or hypothyroidism. The organ-specific autoimmune attack on thyroid witnesses inflammation of the thyroid tissue leading to thyrotoxicosis(21). Also some studies reported that other nonorgan-specific antibodies as anti-dsDNA were not significantly different among patients with AITD and healthy control (22) although our study reveals some significant results regarding different antibodies between cases and control.
In a study conducted on sixty one autoimmune thyroiditis patients free from rheumatic manifestations were assessed for the presence of these non-organ-specific autoantibodies, Regarding ATD patients, anti-ds DNA, were present in a percentage of (18%) with statistically significance difference (P < 0.5) rather than controls, meaning this result agree with our result (19).
Table (2) Comparison between Autoimmune Thyroiditis patients group (n=60) and Healthy Control group (n=30) according to Test Results (AMA and Anti-ds DNA).
Parameters Groups Mean ±SD P-Value
AMA Autoimmune
Thyroiditis
6.15±3.25 0.0001 I*
Control 2.73±2.01
Anti-ds DNA Autoimmune Thyroiditis
0.98±0.63 0.0008 **
Control 0.34±0.14
* (P≤0.05) - Significant, ** (P≤0.01)-Highly Significant.
Conclusion: AITD in Iraq affect females more than males. The patients with AITD are prone for other autoimmune disease. Anti-dsDNA antibody can be used to determine thyroid autoimmunity.
References:
1. Armstrong M, Asuka E, Fingeret A. Physiology, thyroid function. StatPearls [https://www.ncbi.nlm.nih.gov/books/NBK537039]. 2020.
2. Guo Q, Wu Y, Hou Y, Liu Y, Liu T, Zhang H, et al. Cytokine secretion and pyroptosis of thyroid follicular cells mediated by enhanced NLRP3, NLRP1, NLRC4, and AIM2 inflammasomes are associated with autoimmune thyroiditis. Frontiers in immunology. 2018;9:1197.
3. Beradhi SB, Flesch B, Hansen M, Matheis N, Kahaly G. HLA class II differentiates between thyroid and polyglandular autoimmunity. Hormone and Metabolic Research. 2016;48(04):232-7.
4. Medenica S, Garalejic E, Arsic B, Medjo B, Bojovic Jovic D, Abazovic D, et al. Follicular fluid thyroid autoantibodies, thyrotropin, free thyroxine levels and assisted reproductive technology outcome. Plos one. 2018;13(10):e0206652.
5. Diana T, Olivo PD, Kahaly GJ. Thyrotropin receptor blocking antibodies. Hormone and Metabolic Research. 2018;50(12):853.
6. Rekvig OP. The dsDNA, Anti-dsDNA antibody, and lupus nephritis: what we agree on, what must be done, and what the best strategy forward could be. Frontiers in immunology. 2019;10:1104.
7. He L, Zhu X, Yang Q, Li X, Huang X, Shen C, et al. Low serum IL-17A in pregnancy during second trimester is associated with an increased risk of subclinical hypothyroidism. Frontiers in Endocrinology. 2020;11:298.
8. VALLAS M, MONIER J, TOURNIAIRE J, DAMON H, GIRARD D. Prévalence des anticorps anti-mitochondries chez les femmes atteintes de thyroïdite de Hashimoto. La Presse médicale (1983).
1994;23(24):1117-20.
9. Calcaterra V, Nappi RE, Regalbuto C, De Silvestri A, Incardona A, Amariti R, et al. Gender differences at the onset of autoimmune thyroid diseases in children and adolescents. Frontiers in endocrinology. 2020;11:229.
10. Ørstavik KH. Why are autoimmune diseases more prevalent in women? Tidsskrift for Den norske legeforening. 2017.
11. Amador-Patarroyo MJ, Rodriguez-Rodriguez A, Montoya-Ortiz G. How does age at onset influence the outcome of autoimmune diseases? Autoimmune diseases. 2012;2012.
12. Al-Hashimi AM. Hashimoto's thyroiditis: a personal experience. Al-Qadisiyah Medical Journal. 2014;10(17):248-52.
13. Hasan RI, Raziq AH. Studying the Frequency of Autoimmune Thyroid Diseases in Duhok Province. Science Journal of University of Zakho. 2019;7(2):45-9.
14. Fairweather D, Frisancho-Kiss S, Rose NR. Sex differences in autoimmune disease from a pathological perspective. The American journal of pathology. 2008;173(3):600-9.
15. AK Al-Irhayim B, F Lazim A. Thyroiditis in Mosul: a clinico-pathological study. Annals of the College of Medicine, Mosul. 2008;34(2):167-75.
16. Zeng Q, Zhao L, Wang C, Gao M, Han X, Chen C, et al. Relationship between autoimmune liver disease and autoimmune thyroid disease: a cross-sectional study. Scandinavian journal of
17. Pedro A, Romaldini J, Americo C, Takei K. Association of circulating antibodies against double-stranded and single-stranded DNA with thyroid autoantibodies in Graves' disease and Hashimoto's thyroiditis patients. Experimental and clinical endocrinology & diabetes.
2006;114(01):35-8.
18. Dua’a Akram A, Hasan BF, AL-Hafidh AH. Estimation of Some Immunological and Biochemical in the Patients with Systemic Lupus Erythematosus in Males and Females in Baghdad.
Medico Legal Update. 2020;20(3):972-8.
19. Elnady BM, Kamal NM, Shaker RH, Soliman AF, Hasan WA, Alghamdi HA, et al. Prevalence and clinical significance of nonorgan specific antibodies in patients with autoimmune thyroiditis as predictor markers for rheumatic diseases. Medicine. 2016;95(38).
20. Chung W-S, Lin C-L, Kao C-H. Comparison of acute respiratory events between asthma–
COPD overlap syndrome and COPD patients: a population-based cohort study. Medicine.
2015;94(17).
21. Rahman S, Archana A, Jan AT, Dutta D, Shankar A, Kim J, et al. Molecular insights into the relationship between autoimmune thyroid diseases and breast cancer: a critical perspective on autoimmunity and ER stress. Frontiers in immunology. 2019;10:344.
22. Benhatchi K, Lazúrová I, Kozáková D, Rovenský J. Prevalence of rheumatic manifestations and non-organ specific autoimmunity in patients with autoimmune thyreopathy. Vnitrni lekarstvi.
2010;56(2):106-10.
