Home About us Editorial board Search Ahead of print Current issue Archives Submit article Instructions Subscribe Contacts Login 


 
 Table of Contents  
ORIGINAL ARTICLE
Year : 2017  |  Volume : 30  |  Issue : 4  |  Page : 1244-1249

The clinical significance of serum anti-heat-shock protein 27 antibody levels in β-thalassemia patients


1 Department of Pediatrics, Faculty of Medicine, Menoufia University, Shebeen El-Kom, Egypt
2 Department of Clinical and Chemical Pathology, Faculty of Medicine, Menoufia University, Shebeen El-Kom, Egypt
3 Department of Pediatrics, Damanhur Medical Institute, Damanhur, Egypt

Date of Submission03-Apr-2017
Date of Acceptance15-May-2017
Date of Web Publication04-Apr-2018

Correspondence Address:
Shereen M El-Shazly
Department of Pediatrics, Damanhur Medical Institute, Damanhur, El-Beheira
Egypt
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/mmj.mmj_244_17

Rights and Permissions
  Abstract 


Objective
The aim of this study was to measure serum anti-heat-shock protein 27 (HSP27) antibody levels in β-thalassemia patients and to assess the potential associations with the clinical and laboratory characteristics in these patients.
Background
β-Thalassemia syndromes are one of the most common quantitative hemoglobinopathies. HSP27 is a protein that is produced by the cells as a result of exposure to oxidative stress. Anti-HSP27 antibodies are formed as a result of autoimmune response against HSP27 and they are responsible for the pathogenesis of various disorders.
Patients and methods
In this study, we analyzed serum anti-HSP27 antibody levels in 60 β-thalassemia patients (30 β-thalassemia major and 30 β-thalassemia intermedia patients) who were recruited from the Pediatric Department, Menoufia University Hospitals, in the period between January 2016 and May 2016. In addition, 30 healthy children matched for age and sex were included as controls.
Results
Significantly higher serum levels of anti-HSP27 antibodies were found in β-thalassemia patients compared with the controls (P < 0.001). The levels of anti-HSP27 antibodies showed a significantly positive correlation with serum ferritin (r = 0.568; P < 0.001) and serum bilirubin (r = 0.321; P = 0.01) and a significantly negative correlation with age at first diagnosis of thalassemia (r=−0.763; P < 0.001) and the interval of blood transfusions (r=−0.775; P < 0.001).
Conclusion
Serum anti-HSP27 antibody may be a useful biomarker of oxidative stress in patients with β-thalassemia.

Keywords: anti-heat-shock protein 27, oxidative stress, β-thalassemia


How to cite this article:
Ragab SM, El-Hawy MA, Shehata AM, El-Shazly SM. The clinical significance of serum anti-heat-shock protein 27 antibody levels in β-thalassemia patients. Menoufia Med J 2017;30:1244-9

How to cite this URL:
Ragab SM, El-Hawy MA, Shehata AM, El-Shazly SM. The clinical significance of serum anti-heat-shock protein 27 antibody levels in β-thalassemia patients. Menoufia Med J [serial online] 2017 [cited 2024 Mar 28];30:1244-9. Available from: http://www.mmj.eg.net/text.asp?2017/30/4/1244/229211




  Introduction Top


Thalassemia syndromes are inherited hemoglobin disorders that result from decreased or absent production of normal globin chains. This leads to defective globin chains' accumulation, premature red blood cell death, and anemia. They are the most common recessive disorders worldwide[1].

β-Thalassemias disorders can be classified according to the severity of the clinical manifestations into β-thalassemia minor, β-thalassemia major (β-TM), and β-thalassemia intermedia (β-TI), which lies between β-thalassemia minor and β-TM[2],[3].

Disturbance in the balance between oxidants and reducing agents within the body leads to the development of oxidative stress, which has deleterious effects on the cells and tissues in the body. Repeated blood transfusions in β-thalassemia major patients induce oxidative stress as a result of excess free-radical intermediates as well as the shortage of antioxidant capacity[4].

Iron overload is considered one of the most common β-thalassemia complications that results from frequent transfusions and increased gastrointestinal iron absorption[5]. Despite improved survival with chelation therapy, cardiac and vascular complications are still relatively common among β-TM patients and can be explained by both a hypercoagulability and an impairment of vascular endothelial function[6].

Heat-shock proteins (HSPs) are a group of molecular chaperones that correct folding of partially folded or denatured proteins and play a vital role in supporting the capability of cells to overcome stressful and undesirable environmental conditions[7].

Heat-shock protein 27 (HSP27) is described as a member of the small HSP family that maintains cell survival under stressful conditions by handling of proteins of abnormal folding and prevention of cell death. Also, HSP27 is involved in new blood vessel formation, tumor cell expansion, and cytoskeleton reconstruction[8].

HSP27 and anti-HSP27 have been linked to several diseases by acting as a protective or a precipitating factor. It was found that overexpression of HSP27 protects cardiac myocytes against ischemic injury; in contrast, elevated anti-HSP27 titers may be a risk factor for atherosclerosis and cardiovascular complications[9].

The aim of this study was to investigate serum anti-HSP27 antibody levels in β-thalassemia patients and to estimate the potential associations with the clinical and laboratory characteristics in these patients.


  Patients and Methods Top


The present study included 60 (32 males and 28 females; their ages ranged from 1.5 to 10 years) patients diagnosed with β-thalassemia and recruited from the Department of Pediatrics, Menoufia University Hospitals, between January 2016 and May 2016. All patients were proven to have β-thalassemia on the basis of clinical and laboratory findings. In addition, 30 healthy children were included as a control group.

The study was designed to include three groups: group I included 30 patients with β-TM (15 males and 15 females; their ages ranged between 1.5 and 10 years). Group II included 30 (17 males and 13 females; their ages ranged between 4 and 10 years) patients with β-TI. Group III included 30 (12 males and 18 females; their ages range from 3 to 10 years) children as control group.

For all participants, a full assessment of history, clinical examination, assessment of anthropometric parameters, and laboratory investigations were performed.

The Ethical Committee of the Faculty of Medicine, Menoufia University approved this study. The guardian of each patient or control provided written informed consent.

Analytical procedures

Blood samples for laboratory tests were withdrawn by sterile venipuncture. For all patients and controls, the following laboratory tests were performed: complete blood count was analyzed using an automated CELL-DYN Ruby hematology analyzer (Abbott, Chicago, Illinois, USA); liver and kidney function tests were performed using an automated AU480 Analyzer (Beckman Coulter Inc., Brea, California, USA); and serum ferritin levels were measured using a mini VIDAS immunoanalyzer (Biomerieux, Marcy-I'Etoile, France).

Serum samples for the estimation of anti-HSP27 antibody levels were kept frozen at –20°C until analysis. Serum HSP27 autoantibodies (anti-HSP27 IgG) were measured using a specific enzyme-linked immunosorbent assay kit (catalog no. EH7001-1; Assaypro, St Charles, Missouri, USA) according to the manufacturer's instructions.

Statistical analysis

Data were expressed in the form of range, mean, SD, and median for continuous quantitative variables, whereas number and percent were used to describe of qualitative variables. Categorical data were compared using the χ2-test. The Kolmogorov–Smirnov test was used to verify the normality of distribution. Analysis of variance and Kruskal–Wallis tests were used to compare multiple groups of normally and abnormally distributed quantitative data, respectively. For comparison of normally and abnormally distributed quantitative data for two groups, the Student t-test and the Mann–Whitney U-test were used, respectively. Spearman's correlation test was used to examine possible correlations between variables. All analyses were carried out using the statistical package for social science (IBM SPSS, version 20; IBM Corp., Armonk, New York, USA). A two-sided P value less than 0.05 was considered statistically significant in all analyses.


  Results Top


The 90 children included in our study were categorized into 30 β-TM patients, 30 β-TI patients, and 30 apparently healthy children.

A significant difference in consanguinity was found between thalassemia major and thalassemia intermedia groups (P = 0.028). The age at first diagnosis of thalassemia was significantly earlier in the thalassemia major group compared with the thalassemia intermedia group (P< 0.001) and the interval between blood transfusions was significantly longer in the thalassemia intermedia group than in the thalassemia major group (P< 0.001) [Table 1].
Table 1: Comparison between the different groups studied according to demographic and clinical data

Click here to view


The hemoglobin concentration was significantly lower in the thalassemia major and intermedia groups compared with the control group (P< 0.001). Aspartate aminotransferase was significantly higher in the thalassemia major and intermedia groups than in the control group (P< 0.001) [Table 2].
Table 2: Comparison between the different groups studied according to different laboratory parameters

Click here to view


Significantly higher concentrations of serum bilirubin and ferritin were observed in the thalassemia major and intermedia groups compared with the control group (P< 0.001) and also serum bilirubin and ferritin concentrations were significantly higher in the thalassemia major group than in the thalassemia intermedia group (P = 0.011, <0.001, respectively) [Table 2].

Anti-HSP27 antibody levels were significantly higher in thalassemia major patients compared with thalassemia intermedia patients (P< 0.001); also, significantly higher concentrations of serum anti-HSP27 antibodies were found in thalassemia major and intermedia groups compared with the control group (P< 0.001) [Table 2] and [Figure 1].
Figure 1: Comparison between the different groups studied according to heat-shock protein 27 antibodies.

Click here to view


Anti-HSP27 antibody levels showed no significant association with age, height, weight, aspartate aminotransferase, creatinine, and urea. Serum anti-HSP27 antibodies showed a significant positive correlation with serum ferritin (r = 0.568; P < 0.001) [Figure 2] and serum bilirubin (r = 0.321; P = 0.01). Furthermore, inverse correlations were found between anti-HSP27 antibody levels and age at first diagnosis of thalassemia (r=−0.763; P < 0.001) and the interval of blood transfusions (r=−0.775; P < 0.001) [Figure 3] and [Table 3].
Table 3: Correlation between serum heat-shock protein 27 antibody levels and different parameters in β-thalassemia patients and in controls

Click here to view
Figure 2: Correlations between heat-shock protein 27 antibody level and serum ferritin.

Click here to view
Figure 3: Correlations between heat-shock protein 27 antibody level and the interval of blood transfusion.

Click here to view



  Discussion Top


β-Thalassemia manifestations are the result of excess unpaired α-globin chains and excess body iron load that enhance oxidative stress and thus premature death of red blood cells. Ultimately, anemia and several complications occur in those patients. Iron plays a vital role in multiple metabolic pathways in the body, but excess iron load can promote the production of toxic radicals and deleterious complications[10].

Multiple oxidative stress markers were assessed previously in many studies; however, in the current study, we chose anti-HSP27 antibodies in β-thalassemia patients on the basis of the previous knowledge that HSP27 is produced by cells in response to stressful conditions. Our aim was to focus on the oxidative stress burden in thalassemic patients and its possible associations with the clinical and laboratory findings in these patients.

In our study, a statistically significant increase in serum anti-HSP27 antibody levels was found in β-thalassemia patients compared with healthy controls. These results were in agreement with the study of Ghahremanlu et al.[11], which investigated the levels of various oxidative stress markers including anti-HSP27 antibodies in thalassemia major patients.

β-TM patients require chronic blood transfusions, whereas β-TI patients are not dependent on blood transfusion, but ultimately, they suffer from excess iron load because of excess iron absorption. As iron load increases in the body, the transferrin potency to bind iron is impaired and thus serum nontransferrin bound iron levels increase and promote the production of reactive species of oxygen. Organ damage and other diverse complications in β-thalassemia patients are caused by the harmful effects of such free radicals[12].

In the present study, anti-HSP27 antibody levels showed no significant association with age; this was not in agreement with a study by Ghahremanlu et al.[11], in which an inverse correlation was found between anti-HSP27 antibody levels and the age of thalassemic patients. Our findings may be attributed to the differences in the age distribution between our study and Ghahremanlu et al.[11] study (age range of 1.5–10 vs. 7–21 years, respectively). The study of Ghahremanlu et al.[11] included both children and adolescents and they explained their results by the anticipated improvement of defense mechanisms against oxidative stresses during adolescence.

In our study, we found significant inverse associations between anti-HSP27 antibody levels and age at first β-thalassemia diagnosis and the interval between blood transfusions. Again, these findings were not in agreement with the results reported by Ghahremanlu et al.[11]. Our findings can be attributed to the more elevated serum anti-HSP27 antibody levels, greater cellular stress and premature death of red blood cells, and thus the greater need for frequent blood transfusion.

The discrepancies between the results of both studies may be explained by the difference in age and racial groups, the design, and sample size of each study.

The present study proved a positive association between serum anti-HSP27 antibody levels and serum ferritin and bilirubin concentrations. These results were in agreement with Rasool et al.[13], who studied the influence of increased iron load in β-thalassemia patients and found a significant positive association between serum ferritin levels and serum malondialdehyde, which is one of the lipid peroxidation products and can be considered a marker of oxidative stress.

Excess iron load and iron-induced oxidative stress and their deleterious effects in patients with thalassemia major have been investigated frequently [14–18]. Many studies showed that β-thalassemia patients have increased blood levels of labile iron, nontransferrin bound iron, and ferritin [19–21]. Moreover, an overall decrease in antioxidant capacity and an increase in the products of oxidative lipid degradation were documented in these patients [16–18]. Finally, these findings proved that β-thalassemia patients are affected by considerable oxidative stress, which is induced by excess iron.

The positive correlation between anti-HSP27 antibody levels and serum bilirubin can be explained by the increase in oxidative stress burden caused by the absence or a decrease in β-globin biosynthesis. Accumulation of excess unpaired alpha globin chain leads to marked destruction of erythroid cells in the marrow and ineffective erythropoiesis, which results in increased level of serum bilirubin[22],[23].


  Conclusion Top


Serum anti-HSP27 antibody levels were significantly increased in β-thalassemia patients compared with the control group and showed significant correlations with age at first diagnosis of thalassemia, the interval of blood transfusions, and serum ferritin.

Serum anti-HSP27 antibody levels may be a useful biomarker of oxidative stress in β-thalassemia patients.

Further extended studies on anti-HSP27 antibodies using larger numbers of patients and controls are required to clarify their possible role in the development of dangerous complications of β-thalassemia such as cardiomyopathy.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Rund D, Rachmilewitz E. Beta-thalassemia. New Engl J Med 2005; 353:1135–1146.  Back to cited text no. 1
    
2.
Haddad A, Tyan P, Radwan A, Mallat N, Taher A. β-Thalassemia intermedia: a bird's-eye view. Turk J Haematol 2014; 31:5–16.  Back to cited text no. 2
    
3.
Taher AT, Musallam KM, Karimi M, El-Beshlawy A, Belhoul K, Daar S, et al. Overview on practices in thalassemia intermedia management aiming for lowering complication rates across a region of endemicity: the OPTIMAL CARE study. Blood 2010; 115:1886–1892.  Back to cited text no. 3
    
4.
Pavlova LE, Savov VM, Petkov HG, Charova IP. Oxidative stress in patients with beta-thalassemia major. Prilozi 2007; 28:145–154.  Back to cited text no. 4
    
5.
Hahalis G, Alexopoulos D, Kremastinos DT, Zoumbos NC. Heart failure in beta-thalassemia syndromes: a decade of progress. Am J Med 2005; 118:957–967.  Back to cited text no. 5
    
6.
Hahalis G, Kremastinos DT, Terzis G, Kalogeropoulos AP, Chrysanthopoulou A, Karakantza M et al. Global vasomotor dysfunction and accelerated vascular aging in β-thalassemia major. Atherosclerosis 2008; 198:448–457.  Back to cited text no. 6
    
7.
Kumar R, Kumari B, Kumar M. PredHSP: sequence based proteome-wide heat shock protein prediction and classification tool to unlock the stress biology. PLoS One 2016; 11:e0155872.  Back to cited text no. 7
    
8.
Schuster C, Akslen LA, Straume O. Expression of heat shock protein 27 in melanoma metastases is associated with overall response to bevacizumab monotherapy: analyses of predictive markers in a clinical phase II study. PLoS One 2016; 11:e0155242.  Back to cited text no. 8
    
9.
Ghahramanlu E, Banihashem A, Mirhossini NZ, Hosseini G, Mostafavi-Toroghi H, Tavallaie S, et al. Effect of zinc supplementation on serum antibody titers to heat shock protein 27 in patients with thalassemia major. Hematology 2014; 19:113–119.  Back to cited text no. 9
    
10.
Ondei Lde S, Estevão Ida F, Rocha MI, Percário S, Souza DR, Pinhel MA, et al. Oxidative stress and antioxidant status in beta-thalassemia heterozygotes. Rev Bras Hematol Hemoter 2013; 35:409–413.  Back to cited text no. 10
    
11.
Ghahremanlu E, Banihashem A, Saber H, Tavallaie S, Mirhosseini N, Ghayour-Mobarhan M, et al. Increased serum heat shock protein 27 antibody titers and prooxidant-antioxidant balance in patients with beta-thalassemia major. Acta Haematol 2013; 129:1–9.  Back to cited text no. 11
    
12.
Rivella S. Ineffective erythropoiesis and thalassemias. Curr Opin Hematol 2009; 16:187–194.  Back to cited text no. 12
    
13.
Rasool M, Malik A, Jabbar U, Begum I, Qazi MH, Asif M, et al. Effect of iron overload on renal functions and oxidative stress in beta thalassemia patients. Saudi Med J 2016; 37:1239–1242.  Back to cited text no. 13
    
14.
Koren A, Fink D, Admoni O, Tennenbaum-Rakover Y, Levin C. Non-transferrin bound labile plasma iron and iron overload in sickle cell disease: a comparative study between sickle cell disease and β-thalassemic patients. Eur J Haematol 2010; 84:72–78.  Back to cited text no. 14
    
15.
Chakraborty D, Bhattacharyya M. Antioxidant defense status of red blood cells of patients with beta-thalassemia and E/beta-thalassemia. Clin Chim Acta 2001; 305:123–129.  Back to cited text no. 15
    
16.
Kassab-Chekir A, Laradi S, Ferchichi S, Haj Khelil A, Feki M, Amri F, et al. Oxidant, antioxidant status and metabolic data in patients with beta thalassemia. Clin Chim Acta 2003; 338:79–86.  Back to cited text no. 16
    
17.
Walter PB, Fung EB, Killilea DW, Jiang Q, Hudes M, Madden J, et al. Oxidative stress and inflammation in iron overloaded patients with β-thalassaemia or sickle cell disease. Br J Haematol 2006; 135:254–263.  Back to cited text no. 17
    
18.
Livrea MA, Tesoriere L, Pintaudi AM, Calabrese A, Maggio A, Freisleben HJ, et al. Oxidative stress and antioxidant status in beta-thalassemia major: iron overload and depletion of lipid-soluble antioxidants. Blood 1996; 88:3608–3614.  Back to cited text no. 18
    
19.
Sherief LM, Dawood O, Ali A, Sherbiny HS, Kamal NM, Elshanshory M, et al. Premature atherosclerosis in children with beta-thalassemia major: new diagnostic marker. BMC Pediatr 2017; 17:69.  Back to cited text no. 19
    
20.
Porter JB, Walter PB, Neumayr LD, Evans P, Bansal S, Garbowski M, et al. Mechanisms of plasma non-transferrin bound iron generation: insights from comparing transfused diamond blackfan anaemia with sickle cell and thalassaemia patients. Br J Haematol 2014; 167:692–696.  Back to cited text no. 20
    
21.
Cabantchik ZI, Breuer W, Zanninelli G, Cianciulli P. LPI-labile plasma iron in iron overload. Best Pract Res Clin Haematol 2005; 18:277–287.  Back to cited text no. 21
    
22.
De Franceschi L, Bertoldi M, Matte A, Santos Franco S, Pantaleo A, Ferru E, et al. Oxidative stress and β-thalassemic erythroid cells behind the molecular defect. Oxid Med Cell Longev. 2013; 2013:985210.  Back to cited text no. 22
    
23.
Sengsuk C, Tangvarasittichai O, Chantanaskulwong P, Pimanprom A, Wantaneeyawong S, Choowet A et al. Association of iron overload with oxidative stress, hepatic damage and dyslipidemia in transfusion-dependent β-thalassemia/HbE patients. Indian J Clin Biochem 2014; 29:298–305.  Back to cited text no. 23
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2], [Table 3]



 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Patients and Methods
Results
Discussion
Conclusion
References
Article Figures
Article Tables

 Article Access Statistics
    Viewed1908    
    Printed81    
    Emailed0    
    PDF Downloaded125    
    Comments [Add]    

Recommend this journal


[TAG2]
[TAG3]
[TAG4]