Clinical And Experimental Investigation To Co-stimulatory Molecular CD80 In Upregulating Immune Response Of Graves Disease

Objective: To explore various possible factors that maybe influence the prognosis of anti-thyroidism treatment. Method: 96 GD patients were followed by one and half year with regular ATD and then were reexamined for the treatment effect. Finally all patients were classified as remission or relapse group. All patient'serum were collected and TRAb was measured by ELISA, the co-stimulator molecular CD80 mRNA on peripheral blood mononuclear cell membrane was detected by real time PCR and serum total iodine concentration was measured with mild acid digestion method. Those risk factors that may influence the prognosis of GD were selected as independent variable and GD therapeutic effect was selected as dependent variable. Multiple conditional logistics regression study was conducted with the combination of clinical parameters such as serum iodine, TRAb, CD86 mRNA. Outcome: The logistic model determination coefficient R2=0.783(p=0.007) and the risk factors entered into the model were TRAb concentration(βj=0.565,OR=5.506), positive family history(βj=0.528,OR=3.401), CD80 mRNA content(βj=0.453,OR=3.089),serum total iodine concentration(βj=0.389,OR=1.497),age of onset(βj=0.272,OR=1.301)。Conclusion: Those patients that have high level of TRAb, positive family history of GD, increase of expression of CD80 mRNA on PBMC membrane and early onset age were more inclined to relapse after ATD. Abstract Objective: To construct the mouse CD80 expression plasmid vector and express construction of CD80 gene expression in E. coli and DH5αexpression of CD80 protein. Methods: RT-PCR method was Balb/C mice CD80 full length, the recombinant expression plasmid pcDNA3.0-CD80, and transformed into E. coli DH5α, a double digestion and gene sequencing methods screening positive clones, Western blotting to detect expression products. Results: The positive recombinant DH5αin temperature in the CD80 protein expression can be induced by molecular weight consistent with the theoretical value. Lymphocyte transformation test results show that the protein has significant immune-enhancing role. Conclusion: we constructed the CD80 gene expression vector plasmid by genetic engineering technology was in mouse protein for further research on the biological function of mouse CD80 laid the foundation. Purpose: To explore the difference in establishment of Graves disease animal model between gene gun injection and normal plasmid muscle injection. Methods: 45 mice were randomized into four groups and then were immunized with either gene gun gold particles or muscle plasmid. After 4 weeks mice were killed and blood sample were taken to determine FT3,FT4,TSH with RIA, TSAb concentration with Elisa, Spleen was cultured to determine IL-4 concentration with RIA. TRAb activity was calculated with production of cAMP after serum was cultured with hTSHR-CHO cell. Thyroid HE section was applied to observe the thyroid gland histomology. Outcome: Mice immunized with gene gun gold particles compared with other groups after 4th week showed a significant increase in FT4 (0.34±0.11)pg/ml (F=14.34,p=0.002), TRAb activity(167.27±37.37)%(Hc=35.198, p=0.007)and IL-4 production in spleen cell(s0.244±0.092)pmol/L (F=7.0403,p=0.005) than GG0 group in which FT4(0.17±0.08)pg/ml, TRAb activity(98.27±14.80)%and IL-4 production(0.151±0.041)pmol/L and PLS group in which FT4(0.21±0.06)pg/ml, TRAb activity(113.81±65.50)%and IL-4 production(0.173±0.039)pmol/L . TRAb concentration (29.3±1.2)IU/L in gene gun group was higher than GG0 (13.3±0.2)IU/L group(F=9.02,p<0.001)but no difference between PLS (24.3±0.9)IU/L group(q=-1.32,p=0.423). HE sections showed that mice immunized successfully with gene gun had thyroid gland follicular cells hyperplasic ,however, they didn't had any signs of cellular destruction or lymphocyte infiltration. Conclusion: Female Balb/c mice immunized with gene gun gold particles was an feasible way to establish Graves disease animal model, from the second to fourth week expected immunized response could be observed including increase of FT4 and autoimmune antibody.