Abstract
BackgroundB-cell acute lymphoblastic leukemia (B-ALL) is the most common malignancy of childhood. Even though significant progresses have been made in the treatment of B-ALL, some pediatric B-ALL have still poor prognosis. The identification of tumor autoantibodies may have utility in early cancer diagnosis and immunotherapy. In this study, we used serological proteome analysis (SERPA) to screen serum autoantibodies of pediatric B-ALL, aiming to contribute to the early detection of B-ALL in children.MethodsThe total proteins from three pooled B-ALL cell lines (NALM-6, REH and BALL-1 cells) were separated using two-dimensional gel electrophoresis (2-DE), which was followed by Western blot by mixed serum samples from children with B-ALL (n=20) or healthy controls (n=20). We analyzed the images of 2-D gel and Western blot by PDQuest software, and then identified the spots of immune responses in B-ALL samples compared with those in control samples. The proteins from spots were identified using mass spectrometry (MS). The autoantibodies against alpha-enolase (α-enolase) and voltage-dependent anion-selective channel protein 1 (VDAC1) were further validated in sera from another 30 children with B-ALL and 25 normal individuals by the use of enzyme-linked immunosorbent assay (ELISA). The protein expression levels of the candidate antigens α-enolase and VDAC1 in B-ALL were thoroughly studied by immunohistochemical analysis.ResultsUtilizing the SERPA approach, α-enolase and VDAC1 were identified as candidate autoantigens in children with B-ALL. The frequencies of autoantibodies against α-enolase and VDAC1 in children with B-ALL were 27% and 23% by using ELISA analysis, respectively, which were significantly higher than those in normal controls (4% and 0, p<0.05). Immunohistochemical analysis showed the expression of α-enolase and VDAC1 was positive in 95% and 85% of B-ALL patients, respectively, but negative expression levels were showed in the control group.ConclusionsThis study incidated that α-enolase and VDAC1 may be the autoantigens associated with B-ALL. Therefore, α-enolase and VDAC1 autoantibodies may be the potential serological markers for children with B-ALL.
Highlights
B-cell acute lymphoblastic leukemia (B-Acute lymphoblastic leukemia (ALL)) is the most common malignancy of childhood
Identification of multiple proteins from a single gel spot may be due to lower resolution of proteins through broad pH range used for iso-electric focusing in two-dimensional electrophoresis (2-DE)
We demonstrated that positivity for autoantibodies against voltage-dependent anion-selective channel protein 1 (VDAC1) was observed in sera from 7 to 30 (23%) children with ALL, but no such activity was observed in 25 (0%) normal children
Summary
B-cell acute lymphoblastic leukemia (B-ALL) is the most common malignancy of childhood. The identification of tumor autoantibodies may have utility in early cancer diagnosis and immunotherapy. We used serological proteome analysis (SERPA) to screen serum autoantibodies of pediatric B-ALL, aiming to contribute to the early detection of B-ALL in children. Acute lymphoblastic leukemia (ALL) is the most common type of neoplasms in childhood (children aged 0–14 years), and its incidence peaks between the ages of 1 and 4 years [1, 2]. There is an urgent need to find biomarkers which are noninvasive and specific for diagnosis and targeted therapy of pediatric B-ALL. Autoantibodies have been suggested in the serum of patients with a wide variety of tumor and have shown possible for use as biomarkers for tumor diagnosis [6]. The diagnostic specificity of serum p53 antibody may be as high as 96% [11]
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