|Year : 2022 | Volume
| Issue : 2 | Page : 114-120
Association of human leukocyte antigen-G 14-bp insertion/deletion and +3142G>C polymorphisms with rheumatoid arthritis: A meta-analysis
Animesh Chowdhury1, Manoj Lama2
1 Department of Zoology, University of Gour Banga; Department of Zoology, Malda College, Malda, West Bengal, India
2 Department of Zoology, University of Gour Banga, Malda, West Bengal, India
|Date of Submission||21-Dec-2021|
|Date of Acceptance||01-Feb-2022|
|Date of Web Publication||24-May-2022|
Dr Manoj Lama
Molecular Immunology Laboratory, Department of Zoology, University of Gour Banga, Malda - 732 103, West Bengal
Source of Support: None, Conflict of Interest: None
This meta-analysis investigated the significant association between human leukocyte antigen (HLA)-G 14-bp insertion/deletion and +3142G>C polymorphisms with susceptibility to rheumatoid arthritis (RA). This relationship has been scrutinized in several studies, and the findings are controversial. The association studies related to HLA-G gene polymorphisms and RA were extracted from PubMed, Mendeley, ScienceDirect, and Google Scholar databases up to December 31, 2020. Finally, a total of 1516 cases and 1683 control samples were chosen from 7 published research articles for this meta-analysis. A significant association was evaluated using the pooled odds ratios (ORs) and 95% confidence intervals (CIs) under allelic, dominant, recessive, and additive models. Overall, there is absence of significant association between 14-bp ins/del polymorphism and RA (recessive model: OR = 1.012, 95% CI = 0.841–1.217, P = 0.900; dominant model: OR = 1.068, 95% CI = 0.919–1.241, P = 0.393; ins/ins vs. del/del: OR = 1.059, 95% CI = 0.861–1.303, P = 0.587). In the case of +3142G>C polymorphism, the recessive model (OR = 1.332, 95% CI = 1.083–1.640, P = 0.007) showed a significant association with RA susceptibility. When subgroup analysis was done by ethnicity, +3142G>C polymorphism was found to be significantly associated with RA susceptibility in the Asian population under the recessive model (OR = 1.450, 95% CI = 1.048–2.006, P = 0.025). This meta-analysis brings to light that there was no significant association between 14-bp ins/del polymorphism and RA susceptibility. Whereas, +3142G>C polymorphism was found to be significantly associated with susceptibility to RA.
Keywords: +3142G>C, 14-bp ins/del, human leukocyte antigen-G, meta-analysis, polymorphism, rheumatoid arthritis
|How to cite this article:|
Chowdhury A, Lama M. Association of human leukocyte antigen-G 14-bp insertion/deletion and +3142G>C polymorphisms with rheumatoid arthritis: A meta-analysis. Indian J Health Sci Biomed Res 2022;15:114-20
|How to cite this URL:|
Chowdhury A, Lama M. Association of human leukocyte antigen-G 14-bp insertion/deletion and +3142G>C polymorphisms with rheumatoid arthritis: A meta-analysis. Indian J Health Sci Biomed Res [serial online] 2022 [cited 2022 Jul 2];15:114-20. Available from: https://www.ijournalhs.org/text.asp?2022/15/2/114/345831
| Introduction|| |
Rheumatoid arthritis (RA), a chronic inflammatory disorder, is characterized by polyarthritis with often progressive joint damage, immunologic abnormalities, systemic inflammation, increased comorbidity, and premature mortality. It causes severe inflammation, swelling of affected body parts, commonly in the hands, wrists, and knees. In addition, bone erosion and joint deformities can result from the associated inflammation of RA for a long time. In females, this disease is 2–3 times more common than in males affecting approximately 1% of the human population worldwide. To date, RA pathogenesis has the involvement of several genes or genetic loci. A significant genetic region identified in RA pathogenesis is the human leukocyte antigen (HLA) locus, which leads to about half of the genetic susceptibility to RA known so far.,
HLA-G is a nonclassical class Ib major histocompatibility complex located on chromosome 6p21.31. Low levels of polymorphism characterize the HLA-G gene. The IMGT-HLA update 3.35.0, January 2019, includes 61 HLA-G alleles, 19 HLA-G proteins, and 3 null alleles. HLA-G gene contains a 14-bp insertion/deletion (rs66554220) and a +3142G>C (rs1063320) polymorphism in 3' untranslated region (UTR). The 3' UTR becomes apparent to play an important role in regulating HLA-G expression, mainly through posttranscriptional regulatory mechanisms. A crucial role is played by HLA-G in immune suppression and decreasing cytotoxicity of natural killer (NK) cells and CD4 + and CD8 + lymphocytes. The HLA-G-specific polymorphism tends to influence its frequency of expression and may affect its vulnerability to autoimmune disorders. HLA-G works by interacting with inhibitory receptors (ILT2, ILT4, KIR2DL4, CD8, and CD160) expressed on the surface of effector immune cells, including subset populations of T-cells, B-cells, NK cells, and antigen-presenting cells (APCs). HLA-G 14-bp insertion/deletion polymorphisms affect the HLA-G function by influencing the stability and splicing pattern of the HLA-G mRNA isoform. The HLA-G +3142G allele has a high-affinity binding site for microRNAs, which reduces HLA-G expression.
Several studies have focused on the interaction of multiple polymorphisms in 3' UTR and promoter regions of the HLA-G gene with RA. Wu et al. failed to find any HLA-G 14-bp insertion/deletion association with autoimmune rheumatic diseases. Rubini et al. suggested that the HLA-G 14-bp insertion/deletion polymorphism affects the likelihood of developing RA but only in females. Hashemi et al. observed a significant association between HLA-G +3142G>C polymorphism and susceptibility to RA in the Iranian population. Veit et al. also suggested an association between an HLA-G 3' UTR +3142G>C polymorphism with susceptibility to RA. Catamo et al. failed to find any association between 14-bp insertion/deletion and +3142G>C polymorphisms and risk for RA in the Brazilian population. HLA-G 3' UTR polymorphisms may have functional significance as the disease-specific role of HLA-G can be related to the breakdown in the resistance or the production of autoimmune disorders, including RA.
To address the shortcomings of individual experiments, meta-analysis is being conducted to fix contradictions and reduce the probability that random mistakes are responsible for positive or false-negative comparisons. Therefore, this meta-analysis and systematic review was conducted to derive a more accurate association of HLA-G 14-bp insertion/deletion and +3142G>C polymorphisms with RA susceptibility.
| Materials and Methods|| |
Identification of eligible studies
The PRISMA guidelines were followed when conducting this study (Preferred Reporting Items for Systemic Assessments and Meta-Analyses). PubMed (www.ncbi.nlm.nih.gov/pubmed), Mendeley (www.data.mendeley.com), ScienceDirect (www.sciencedirect.com), and Google Scholar (www.scholar.google.com) databases were searched thoroughly for all eligible studies on 14-bp insertion/deletion and +3142G>C polymorphism in RA (up to December 31, 2020). We searched and included all potential studies using different combinations of keywords, namely 14 bp insertion-deletion polymorphism in rheumatoid arthritis or rheumatoid arthritis 14 bp ins del polymorphism or 14 bp ins del polymorphism in RA or +3142G>C polymorphism in rheumatoid arthritis or 3' UTR polymorphism in rheumatoid arthritis.
Inclusion and exclusion criteria
In compliance with inclusion and exclusion requirements, all the articles found through database searching have been screened. In addition, all studies which were not considered significant for this meta-analysis were excluded. The inclusion criteria were (i) presence of original data containing 14-bp insertion/deletion polymorphism in RA, (ii) +3142G>C polymorphism in RA, (iii) case–control studies, and (iv) 3' UTR polymorphism containing G>C polymorphism. However, we excluded (i) review articles on RA, (ii) studies not having 14-bp ins/del or +3142G>C polymorphism, (iii) studies that have incomplete data on 3' UTR polymorphism in RA, and (iv) studies that did not show genotypic frequencies.
Data obtained from the potential articles include (1) first author's name, (2) year of publication, (3) country of origin, (4) genotyping method, (5) number of cases and controls, (6) genotypic frequencies of 14-bp insertion/deletion polymorphism in cases and controls, and (7) frequencies of CC, GC, and GG genotypes in cases and controls. [Table 1] presents the characteristics of the studies considered in this analysis. Two independent authors extracted data from various studies listed from the above databases, and by consensus, potential conflicts were resolved.
The correlation power was measured using a pooled odds ratio (OR) and their 95% confidence intervals (CIs) between HLA-G 14-bp ins/del and +3142G>C polymorphisms in RA susceptibility. The pooled ORs were calculated under following genotyping contrast models: the allele model (ins vs. del), the homozygous model (ins/ins vs. del/del), the heterozygous model (ins/del vs. del/del), the recessive model (ins/ins vs. ins/del + del/del), and the dominant model (ins/ins + ins/del vs. del/del).The significance of the pooled OR with the Z-test was calculated with a statistically significant P < 0.05. The heterogeneity was calculated by Cochran's Q statistics and the I2 test., Heterogeneity was found to be substantial when P < 0.10. A random-effect model was selected when the I2 value is >50%; otherwise, the fixed-effect model was adopted to test the heterogeneity. The Begg's funnel plot and Egger's linear regression test were carried out, and P < 0.05 was found significant to assess the existence of the publication bias., These analyses were performed by STATA version 16.0 (StataCorp LP, College Station, Texas, USA).
| Results|| |
Characteristics of eligible studies
The selection process of potential studies for this meta-analysis is depicted in [Figure 1]. After completion of the search process, a total of 32 articles were initially identified. However, on applying the inclusion and exclusion criteria as a set, 7 publications were included with a total of 1516 cases and 1683 controls for this meta-analysis. Among them, 3 publications were characterized as Asian,,, and 4 were Caucasian.,,, Only 3 published articles were on +3142G>C polymorphism in RA.,, All of the articles were in HWE except for one. The characteristics of all eligible studies are summarized in [Table 1].
|Figure 1: PRISMA flowchart illustrating the search strategy used to identify relevant studies. Fom: Moher D, Liberati A, Tetzlaff J, Altman DG, The PISMA Group. Preferred Reporting Items for Systematic Reviews and Meta- Analyses: The PRISMA Statement. PLoS Med 2009;6:e1000097. doi: 10.1371/journal.pmd1000097|
Click here to view
[Table 2] shows the calculation for the heterogeneity test and association of HLA-G 14-bp ins/del and +3142G>C polymorphism with RA susceptibility. Overall, there was absence of significant association between 14-bp ins/del polymorphism and RA. The models tested were allelic model (ins vs. del – OR = 1.034, 95% CI = 0.934–1.145, P = 0.517), recessive model (ins/ins vs. ins/del + del/del – OR = 1.012, 95% CI = 0.841–1.217, P = 0.900), dominant model (ins/ins + ins/del vs. del/del – OR = 1.068, 95% CI = 0.919–1.241, P = 0.393), ins/ins versus del/del (OR = 1.059, 95% CI = 0.861–1.303, P = 0.587), and ins/del versus del/del (OR = 1.070, 95% CI = 0.910–1.257, P = 0.414). Furthermore, in comparison with respect to ethnicity, there was absence of significant association of HLA-G 14-bp ins/del polymorphism with RA susceptibility in Asian (ins/ins vs. del/del – OR = 0.977, 95% CI = 0.692–1.380, P = 0.896) and Caucasian (ins/ins vs. del/del – OR = 1.108, 95% CI = 0.855–1.435, P = 0.437) populations [Figure 2].
|Table 2: Meta-analysis of associations between the human leukocyte antigen-G 14-bp I/D and +3142G/C polymorphisms in rheumatoid arthritis|
Click here to view
|Figure 2: Forest plot of HLA-G 14bp ins/del polymorphism and RA susceptibility in subgroup analysis by ethnicity using a fixed-effect model (ins/ins vs. del/del)|
Click here to view
In the case of association between HLA-G +3142G>C polymorphism and RA susceptibility, the overall significant association was observed only under the recessive model, i.e., GG versus GC + CC (OR = 1.332, 95% CI = 1.083–1.640, P = 0.007). While no associations were observed under other models: allelic model (G vs. C – OR = 1.178, 95% CI = 0.862–1.611, P = 0.302), dominant model (GG + GC vs. CC – OR = 0.983, 95% CI = 0.456–2.120, P = 0.965), GG vs. CC (OR = 1.159, 95% CI = 0.532–2.524, P = 0.710), and GC vs. CC (OR = 0.819, 95% CI = 0.395–1.702, P = 0.593). When subgroup analysis was done with respect to ethnicity, a marginal association of HLA-G +3142G>C polymorphism with RA was observed in Asian population under recessive model (GG vs. GC + CC – OR = 1.450, 95% CI = 1.048–2.006, P = 0.025) [Table 2] and [Figure 3]. However, no association was found in the case of other models: allelic model (G vs. C – OR = 1.230, 95% CI = 0.645–2.343, P = 0.529), dominant model (GG + GC vs. CC – OR = 0.982, 95% CI = 0.177–5.455, P = 0.983), GG versus CC model (OR = 1.160, 95% CI = 0.209–6.425, P = 0.865), and GC versus CC (OR = 0.760, 95% CI = 0.159–3.638, P = 0.731) in Asian population. Similarly, no significant association of HLA-G +3142G>C polymorphism with RA was observed in Caucasian population under any of the models tested: allelic model (G vs. C – OR = 1.111, 95% CI = 0.932–1.325, P = 0.237), recessive model (GG vs. GC + CC – OR = 1.257, 95% CI = 0.960–1.647, P = 0.097), dominant model (GG + GC vs. CC – OR = 1.026, 95% CI = 0.754–1.397, P = 0.870), GG versus CC model (OR = 1.197, 95% CI = 0.839–1.709, P = 0.322), and GC versus CC (OR = 0.934, 95% CI = 0.673–1.295, P = 0.681).
|Figure 3: Forest plot of HLA-G +3142 G>C polymorphism and RA susceptibility in subgroup analysis by ethnicity using a random-effect model (GG vs. CC)|
Click here to view
No heterogeneity was found in HLA-G 14-bp ins/del polymorphism as Cochran's Q statistics P value is greater than 0.10 (P < 0.10 is significant). However, in the case of +3142G>C polymorphism except for the recessive model (GG vs. GC + CC), all other models, namely allelic model (G vs. C), dominant model (GG + GC vs. CC), and GG versus CC, showed the presence of significant among publications as Cochran's Q statistics P value is lower than 0.10.
Begg's funnel test and Egger's linear regression test were carried out to find out the publication bias of all comparison models. Begg's funnel test showed symmetrical plot [Figure 4], which indicates the absence of publication bias. Egger's linear regression test also showed no significant evidence for publication bias (ins vs. del: P = 0.857, ins/ins vs. del/del: P = 0.584, ins/del vs. del/del: P = 0.430, ins/ins vs. ins/del + del/del: P = 0.315, and ins/ins + ins/del vs. del/del: P = 0.666).
|Figure 4: Begger's funnel asymmetry plot of HLA-G 14bp ins/del polymorphism and RA risk under the homozygous contast (ins/ins vs. del/del) model|
Click here to view
| Discussion|| |
HLA-G plays a critical role in immunosuppression by repressing the cytotoxicity of NK and T-cells (CD8+ and CD4+) and hindering their development as APCs., In many autoimmune disorders, a variable HLA-G expression has been documented, although HLA-G alleles reveal minimal diversity. The polymorphism of 14-bp insertion/deletion located in position +2960 at exon 8 (rs1704) has attracted attention due to its potential role in alternative splicing and RNA stability. The transcriptions with the 14-bp insertion sequence have been shown to undergo an additional splicing step that removes 92 bp from the region in which the sequence is located. The deletion allele stabilizes the mRNA with a consequent higher HLA-G expression. The +3142G> C polymorphism influences the affinity of HLAG mRNA targeted by different microRNAs, as demonstrated by an in silico study. +3142G allele has a binding site with a higher affinity for miR-148a, miR-148b, and miR-152, downregulating the expression of HLA-G., The HLA-G polymorphism appears to change its degree of expression and may influence illness susceptibility in autoimmune diseases.
The current study examined the connection between HLA-G 14-bp ins/del and +3142G>C polymorphism with RA susceptibility. A meta-analysis was performed with 7 eligible studies comprising 1516 cases and 1683 control samples for 14-bp ins/del polymorphism, and 873 cases and 772 control samples for +3142G>C polymorphism. Our data suggest that there is no significant association between 14-bp ins/del polymorphism and RA susceptibility. Whereas, for +3142G>C polymorphism, the only recessive model showed a significant association with susceptibility to RA. In the ethnicity-based subgroup analysis, there was a significant association between +3142G>C polymorphism and RA susceptibility in the Asian population under the recessive model. In heterogeneity analysis, huge contrast was observed under the recessive and allelic models in our meta-analysis. To find out publication bias, we also performed Begg's funnel test and Egger's regression test. The results of both tests confirmed no publication bias in the studies included in the analysis.
In fact, there are a limited number of association studies concerning HLA-G gene polymorphisms and susceptibility to RA, and the findings are found to be contradictory. The only meta-analysis conducted so far to investigate the associations of the functional HLA-G 14-bp insertion/deletion and +3142G>C polymorphisms with susceptibility to RA in addition to systemic lupus erythematosus (SLE) reported no association between these variants of the HLA-G gene and RA. However, ins/ins + ins/del genotype of HLA-G 14-bp ins/del polymorphism and +3142G allele was significantly associated with SLE. One of the studies reported that +3142GG genotype significantly increased the risk of RA (OR = 1.45, 95% CI = 1.075–1.95, P = 0.030). In contrast, the studies have demonstrated that HLA-G +3142G>C polymorphism significantly decreased the risk of RA.,
The findings indicate that the association between HLA-G 14-bp ins/del and +3142G>C polymorphisms and RA susceptibility was not strong enough. This study has some limitations that may be taken into account. First, in this meta-analysis, data of only Asian and Caucasian ethnic groups are subjected to analysis. The general importance of HLA-G polymorphisms during the advancement of RA may rely upon ethnicity. Therefore, due to limited ethnicity-based data, we were incapable of performing ethnicity-specific meta-analyses for different populations. Second, in addition to disease susceptibility, HLA-G polymorphisms may be linked to clinical manifestations of the disease. Third, unadjusted estimates were included in this systematic review. We did not analyze gender, age, clinical manifestations, and environmental factors as all these information were unavailable in the original publications. Fourth, the analysis included a minimal number of studies, and therefore, our findings should be interpreted carefully.
| Conclusion|| |
The results of this meta-analysis exert that there was no significant association between 14-bp ins/del polymorphism and RA susceptibility. However, HLA-G +3142G>C polymorphism showed a significant association with RA susceptibility under the recessive model. A sufficiently large number of association studies on HLA-G gene polymorphisms and susceptibility to RA are required for analysis to arrive at a more tangible conclusion.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Turesson C, Matteson EL. Genetics of rheumatoid arthritis. Mayo Clin Proc 2006;81:94-101.
Iebba F, Di Sora F, Tarasi A, Leti W, Montella F. Rheumatoid arthritis: A typical multifactorial genetic disease: Review of the literature. Recent Prog Med 2011;4:175-82.
Veit TD, De Lima CP, Cavalheiro LC, Callegari-Jacques SM, Brenol CV, Brenol JC, et al
. HLA-G+3142 polymorphism as a susceptibility marker in two rheumatoid arthritis populations in Brazil. Tissue Antigens 2014;83:260-6.
Gautam S, Kumar U, Kumar M, Kanga U, Dada R. Association of HLA-G 3'UTR polymorphisms with soluble HLA-G levels and disease activity in patients with rheumatoid arthritis: A case-control study. Immunol Investig 2020;49:88-105.
Hashemi M, Sandoughi M, Fazeli SA, Bahari G, Rezaei M, Zakeri Z. Evaluation of HLA-G 14 bp Ins/Del and+3142G> C Polymorphism with Susceptibility and Early Disease Activity in Rheumatoid Arthritis. Adv Med 2016;2016:4985745.
Shiroishi M, Kuroki K, Rasubala L, Tsumoto K, Kumagai I, Kurimoto E, et al
. Structural basis for recognition of the nonclassical MHC molecule HLA-G by the leukocyte Ig-like receptor B2 (LILRB2/LIR2/ILT4/CD85d). Proc Natl Acad Sci U S A 2006;103:16412-7.
Rousseau P, Le Discorde M, Mouillot G, Marcou C, Carosella ED, Moreau P. The 14 bp deletion-insertion polymorphism in the 3′ UT region of the HLA-G gene influences HLA-G mRNA stability. Hum Immunol 2003;64:1005-10.
Tan Z, Randall G, Fan J, Camoretti-Mercado B, Brockman-Schneider R, Pan L, et al
. Allele-specific targeting of microRNAs to HLA-G and risk of asthma. Am J Hum Genet 2007;81:829-34.
Wu F, Wu L, Luo X, Tang Z, Yang M, Xia Y, et al
. Association of HLA-G 14bp insertion/deletion polymorphism with autoantibody production in patients with autoimmune rheumatic diseases. J Clin Immunol Immunopathol Res 2010;2:15-21.
Rubini M, Roversi EB, Aiello V, Khan MF. Gender-dependent association between HLA-G 14bp insertion/deletion polymorphism and rheumatoid arthritis in Italian patients. Genomics Genetics Epigenetics Rheumatic Dis 2011;74 (Suppl 2):895-6.
Catamo E, Addobbati C, Segat L, Sotero Fragoso T, Domingues Barbosa A, Tavares Dantas A, et al
. HLA-G gene polymorphisms associated with susceptibility to rheumatoid arthritis disease and its severity in Brazilian patients. Tissue Antigens 2014;84:308-15.
Moher D, Liberati A, Tetzlaff J, Altman DG, PRISMA Group. Preferred reporting items for systematic reviews and meta-analyses: The PRISMA statement. PLoS Med 2009;6:e1000097.
Grant J, Hunter A. Measuring inconsistency in knowledgebases. J Intell Inform Syst 2006;27:159-84.
Higgins JP, Thompson SG. Quantifying heterogeneity in a meta-analysis. Stat Med 2002;21:1539-58.
Begg CB, Mazumdar M. Operating characteristics of a rank correlation test for publication bias. Biometrics 1994;50:1088.
Egger M, Davey Smith G, Schneider M, Minder C. Bias in meta-analysis detected by a simple, graphical test. BMJ 1997;315:629-34.
Veit TD, Chies JA, Switala M, Wagner B, Horn PA, Busatto M, et al
. The paradox of high availability and low recognition of soluble HLA-G by LILRB1 receptor in rheumatoid arthritis patients. PLoS One 2015;10:e0123838.
Veit TD, Vianna P, Scheibel I, Brenol C, Brenol JC, Xavier RM, et al
. Association of the HLA-G 14-bp insertion/deletion polymorphism with juvenile idiopathic arthritis and rheumatoid arthritis. Tissue Antigens 2008;71:440-6.
Bainbridge DR, Ellis SA, Sargent IL. HLA-G suppresses proliferation of CD4(+) T-lymphocytes. J Reprod Immunol 2000;48:17-26.
Dorling A, Monk NJ, Lechler RI. HLA-G inhibits the transendothelial migration of human NK cells. Eur J Immunol 2000;30:586-93.
Baricordi O, Stignani M, Melchiorri L, Rizzo R. HLA-G and inflammatory diseases. Inflamm Allergy Drug Targets 2008;7:67-74.
Carosella ED, Favier B, Rouas-Freiss N, Moreau P, Lemaoult J. Beyond the increasing complexity of the immunomodulatory HLA-G molecule. Blood 2008;111:4862-70.
Castelli EC, Moreau P, Chiromatzo AO, Mendes-Junior CT, Veiga-Castelli LC, Yaghi L, et al
. In silico analysis of microRNAS targeting the HLA-G 3′ untranslated region alleles and haplotypes. Hum Immunol 2009;70:1020-5.
Manaster I, Goldman-Wohl D, Greenfield C, Nachmani D, Tsukerman P, Hamani Y, et al
. MiRNA-mediated control of HLA-G expression and function. PLoS One 2012;7:e33395.
Lee YH, Bae SC, Song GG. Meta-analysis of associations between functional HLA-G polymorphisms and susceptibility to systemic lupus erythematosus and rheumatoid arthritis. Rheumatol Int 2015;35:953-61.
[Figure 1], [Figure 2], [Figure 3], [Figure 4]
[Table 1], [Table 2]