Antinucleolar Antibody Purification And Target Antigen Screening From Serum Of Children With Immune Thrombocytopenia

Background: Immune thrombocytopenia(ITP)is a common immune disease in children.Our previous study found that a high proportion of children with positive antinucleolar antibodies were ITP patients.AnoA plays an important role in many diseases.However,there is no report about the mechanism of AnoA and its target antigens in the patients with ITP.Objective: Our study was to screen and identify the target antigens recognized by antinucleolar antibodies in serum of children with ITP.Methods: A system was established by using HEP-2 cell line: a system of cell immobilization,antibody binding,antibody elution and neutralization.Using protein immunoprecipitation and mass spectrometry to screen the possible target antigens that bind to AnoA in serum of children with ITP.The target antigens were identified by plasmid overexpression and cellular immunofluorescence.Results: The AnoA in serum of children with ITP was purified successfully.Proteins such as NCL,KRT1,KRT2,KRT10 may be the target antigens of AnoA in serum of children with ITP.It was identified that NCL may not be the target antigen.The roles of proteins such as KRT1,KRT2,KRT10 et should be further verified.Conclusion: These results provide an effective method for the identification of target antigens of AnoA in serum of children with ITP and can help us to have a deeper understanding of the pathogenesis of ITP.