Research Of Mucosal Antibody And Antibody Secreting Cells Of Pseudosciaena Crocea

The paper was conducted on Vibrio harveyi ,Pseudosciaena crocea, Immune polysaccharide and Eriocheri sinensis : using microbiological technology , proteinic technology and immunological technology , study on disease-resistance of Pseudosciaena crocea and Eriocheri sinensis ,purification of serum immunoglobulin, investigation of the mucosal antibody and detection of ASCs. The results were summarized as follow.1. The Large Croaker (Pseudosciaena crocea) were injected with crude lipopolysaccharide (LPS) preparations, outer membrane protein (OMP) of Vibrio harveyi and formalin-killed Vibrio harveyi respectively. The immune response were compared with examing the agglutinating antibody titers, the phagocytic activity and lysozyme activity. After 4 weeks, we did challege tests, compare the relative percentage survival (RPS).The results showed that 3 antigens could caused the Large Croaker (Pseudosciaena crocea) strong immune response. The agglutinating antibody titers,the phagocytic activity and lysozyme activity in the blood were higher than the control group. Meanwhile, we found there were no direct relevance between the agglutinating antibody titers and the relative percentage survival (RPS).2. The purification of serum immunoglobulin in Large Yellow Croaker (Pseudosciaena Crocea) was carried out by using ammonium sulfate precipitation followed by colum chromatography, and then inoculated into the New Zealand white rabbit with Freund complete adjuvant. Rabbit anti- Large Yellow Croaker IgG was also purified and coagulated with horseradish peroxidase (HRP). Establish an Indirect ELISA to detection the antibody caused by Vibrio harveyi.Optimum conditions of indirect ELISA were developed as follows: 15ug/ml of antigen used to coat ELISA plates; 1:80 dilution of serum as detecting samples; 1:3000 dilution of HRP-labeled antibody as detecting regent; antigen were absorbed by overnight incubation at 4°C temperature; this indirect ELISA systems were proved to be sensitive and specific.3. The aim of this study was to investigate the mucosal response in Yellow croaker via different routes: oral administration,intraperitoneal injection and immersion. The antibody level was measured using an indirect ELISA.The results indicated: mucosal antibody occurred earlier, maintained shorter and have low level, but it was different in system antibody. Otherwise, different routes have different results. Immersion may stimulate the mucosal system directly, higher antibody levels were found in bile and mucus than in serum on immersion, which was not the case after intraperitoneal injection. The delayed onset of antibody formation in the mucosal compartment after IP injection might indicate a delayed exchange from the systemic to...