Evaluation of the diagnostic value of FliD, a Helicobacter pylori flagellar protein

Abstract

Novel immunodominant antigens are urgently required for diagnosis and vaccine of Helicobacter pylori(HP). FliD, an important colonization factor was cloned and expressed(rFliD) to evaluate the levels of specific-IgG, IgM and IgA antibodies in the serum of patients by ELISA. Rabbit anti-rFliD polyclonal antibody (pAb) was obtained by subcutaneous injections of rFliD. The rFliD-specific IFN-γ and IL-4 of peripheral blood mononuclear cells and CD4⁺ T cells from human were analyzed by enzyme-linked immunospot and flow cytometry. We found that the levels of rFliD-specific IgG, IgM and IgA were significantly higher in HP-infected-patients compared with healthy controls. IgG, IgM and IgA had diagnostic sensitivities of 92.6, 89.8 and 83.2%; specificities of 91.1, 88.7 and 64.6%; and areas under the receiver operating curves of 0.97, 0.96 and 0.92, respectively. Furthermore, rFliD-pAb was used in the immunohistochemical analysis of gastritis and gastric cancer tissues from patients infected with HP. The levels of rFliD-specific IFN-γ and IL-4 were significantly elevated in HP-infected patients and exhibited a T helper type 1-dominant subtype. These findings indicate that rFliD exhibits high validity as a biomarker in HP diagnosis and may also be a potent antigen for vaccine design due to its high cellular and humoral immune response.