Study On Gene Expression And Immune Response After The Immunization Of Somatostatin DNA Vaccine In Mice

Somatostatin(SS) is one of the most important neuropeptides secreted from hypothalamus nuclei which inhibits the secretion of growth hormone(GH) and depresses the growth.At present our lab has completed the construction job of the somatostatin DNA vaccine and also did some experiments in the clinical trials.In order to explore the mechanism of somatostatin gene vaccine pGS/2SS-M4GFP-asd,the immune response and expression distribution of plasmid by methods including organ imaging,frozen section,immunohistochemistry in the present paper.The aims of this paper were to explore the duration of the immune response,immune response arising site,so as to set up a basis for animal clinical experiments and business applications.The experiments include following contents:1.Anti-somatostatin antibody detectionA total of 54 Balb/C mice with 3-week-old and 90 Kunming mice with 7-week-old,each divided into S;C;P group for immunization,group S immune the pGS/2SS-M4GFP-asd vaccine;group C immune the C500;group P immune the PBS.detection of somatostatin IGg plasma antibody level in different periods results showed that:The S group in Balb/C mice and kunming mice induced higher levels of IgG during the course of sampling than from the control groups,all the results showed that after the immunization of somatostatin can reduce the plasma concentrations of somatostatin,which can play a role in promoting growth.2.Organ fluorescence imagingAfter the M4GFP successfully transfect the PK cell,we immune the mice with the SS gene vaccine C500(pGS/2SS-M4GFP-asd),from 12h later,we collected the organs for fluorescence imaging,the results of the experiment group shows:we can see green fluorescent in the liver,spleen,kidney,muscle tissue in 48 hours after immunization.that means the green fluorescent protein can be use as a biomarkers.3.Organ section expressionAfter 12h of the immunization of recombinant Plasmid(encoded the enhanced green fluorescent protein gene(GFP)) we collected the organs for frozen section detection.Using fluorescence microscopy the green fluorescence in the liver and spleen expression can be observed,while the body autofluorescence of the problem is still a problem.Through the Immunohistochemistry,the SS protein can be seen in the mouse liver,leg muscle,and spleen after5 days through the Immunohistochemistry In this study,we successfully transfer the somatostatin gene into the eukaryotic expression vector,and confirmed by the immunization of somatostatin can induce the stable expression,by regulating their growth hormone,to improving the growth performance.