基于血清蛋白质组学日本血吸虫病诊断标志物的筛选及其免疫应答的分析-screening of diagnostic markers for schistosomiasis japonica based on serum proteomics and analysis of its immune response.docxVIP

of the recombinants and analyzed for T-bet, INF-γ, IL-2, GATA-3, IL-4 and IL-10 using Real-time PCR, and the cultured supernatant were collected for INF-γ, IL-2, IL-4 and IL-10 cytokines detection.ResultsThis serological proteome assay yielded more than 30 immunodominant spots. Ten of these spots were precisely matched with a homologous two-dimensional electrophoresis (2-DE) gel and successfully identified by LC/MS-MS as corresponding to four different proteins. Of these proteins, SjLAP and SjFBPA were successfully expressed, and their recombinant protein products were further applied in the diagnosis of human schistosomiasis japonica using ELISA. The ELISA results revealed sensitivities of 98.1% and 87.8% for acute and chronic schistosomiasis with rSjLAP and 100% and 84.7% with rSjFBPA, whereas the assays showed a specificity of 96.7% with both recombinant proteins. After treatment with praziquantel, the titres of the antibodies against both antigens declined significantly (P0.001) within 6 months post-treatment of praziquantel and were reduced to the control level between 6 to 12 months post-treatment in more than 80% patients sera.Anti-rSjLAP or anti-rSjFBPA IgG antibody was induced significantly higher at two weeks and reached the highest levels at six weeks after the immunization of BALB/c mice with either rSjLAP or rSjFBPA than the control group. And the anti-rSjLAP or anti-rSjFBPA IgG1 antibody was the same as its IgG antibody. However, the anti-rSjLAP or anti-rSjFBPA IgG2a antibody level was not significantly different after immunization. The cytokine profiles showed the higher levels of IL-4 and IL-10 were observed than the PBS control group in supernatants of spleen cells stimulated with rSjLAP or rSjFBPA. There was significant difference in the cytokine profiles of INF-γ and IL-2 between the immunized mice and the control. Furthermore, the Real-time PCR determined rSjLAP or rSjFBPA significantly upregulated the mRNA expressions of GATA-3, IL-4 and IL-10 w