A Novel Method to Identify Autoantibodies against Putative Target Proteins in Serum from beta-Thalassemia Major: A Pilot Study

Afshan Sumera,Nurul H Rutt,Nur Diana Anuar,Hishamshah Ibrahim,Ti-Myen Tan,Abdul Aziz Baba,Ammu Kutty Radhakrishnan,Nur Hafiza Ismail

doi:10.3390/biomedicines8050097

Abstract

Abnormal immune reactivity in patients with beta-thalassemia (beta-thal) major can be associated with poor prognosis. Immunome protein-array analysis represents a powerful approach to identify novel biomarkers. The Sengenics Immunome Protein Array platform was used for high-throughput quantification of autoantibodies in 12 serum samples collected from nine beta-thal major patients and three non-thalassemia controls, which were run together with two pooled normal sera (Sengenics Internal QC samples). To obtain more accurate and reliable results, the evaluation of the biological relevance of the shortlisted biomarkers was analyzed using an Open Target Platform online database. Elevated autoantibodies directed against 23 autoantigens on the immunome array were identified and analyzed using a penetrance fold change-based bioinformatics method. Understanding the autoantibody profile of beta-thal major patients would help to further understand the pathogenesis of the disease. The identified autoantigens may serve as potential biomarkers for the prognosis of beta-thal major.

Highlights

Thalassemia is the most commonly inherited single-gene disorder in the world, with an overall carrier frequency of 2%–25% [1]
The most common mutation in these patients was β-codon 41/42 [-TTCT], and HCLS1 was identified in the majority of cases (Table 1)
Comparing the available patient data with the biomarkers identified in this study, we found that β codon 41/42 [-TTCT] was the most common mutation, which is a frequent mutation type in Chinese β-thal carriers [46]

Summary

Introduction

Thalassemia is the most commonly inherited single-gene disorder in the world, with an overall carrier frequency of 2%–25% [1]. Beta-thalassemia (beta-thal) major is a disorder caused by mutations in the β-globin gene, which can result in total absence of β-globin chains. The deficiency of β-globin chains causes an imbalance in the ratio between α- and β-globin chains within mature and immature erythrocytes [4]. Unpaired α-globin chains are very toxic to erythrocytes, as they can liberate reactive iron and induce oxidative stress to the plasma membrane and proteins within these cells. Excess α-globin chains tend to increase intracellular calcium levels, which exposes phosphatidylserine residues; which can result in early apoptosis in the affected red blood cells (RBC). Beta-thal major manifests by ineffective erythropoiesis, peripheral hemolysis and, eventually, anemia [4]

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