Résumé
Objective:On the basis of the use of chemical methods to establish mouse model of lupus nephritis and its biological identification , we investigate the reverse effect of pathological lesions of B 7-1 human-mouse chimeric antibody blockade against B7/D28 signaling pathway in mice with lupus nephritis model.Methods:Pristane was injected intraperitoneally to 6-week-old female C57BL/6J mice at dose of 0.5 ml per mouse in one go,and urine protein,ANA and renal pathological changes were detected on a monthly basis.Mice whose urine protein content reached ++and ANA fluorescence intensity reached ++were randomly devided into three groups ,five each.Antibody intervention group was sequentially injected with B 7-1-mouse chimeric antibody by orbital venous , positive control group was injected with immunosuppressant CTX , negative control group was injected with isotype control IgG.Urine protein and ANA were also detected on a monthly basis.Mice were sacrificed three months after intervention was executed.Kidney was used for H&E dying , IC detection and electric microscope observation.Results: After four-month Pristane induction , urine protein content of 80%mice reached +-+++,meanwhile,serum ANA fluorescence intensity reached ++-+++.Glomerulonephritis infiltrating cells were observed Mice with urine protein and ANA , glomerular inflammatory cell infiltration , tubular epithelial cell degeneration visible edema ,vascular congestion significantly ,fibrosis.After antibody intervention ,urine protein content in antibody intervention group gradually reduced from ++-+++to ±-+++,ANA ++-+++to +-++,and were significantly different from that in the negative control group ( P<0.01 ).Analysis of kidney H&E dying showed that antibody glomerular infiltration of inflammatory cells in the intervention group and tubular congestion and other symptoms were improved significantly.Immunofluorescence staining indicated that fluorescence intensity of IC was significantly reduced in the antibody intervention group.Electron dense deposits reduction and glomerular basement membrane uniformity were observed in antibody intervention group by electric microscope when compared with the negative control group.Conclusion:B7-1 antibodies could downregulate immune response through inhibiting B 7-1/CD28 signaling pathway , reducing the production of autoantibodies and reversing pathological damage caused by autoimmune response .
Résumé
Objective To establish the sensitive,specific,stable and convenient immunoradio assay for detecting human soluble IL-6R?.Methods The hybridoma cell lines were obtained by fusing spleen cells of BALB/c mice that had been immunized with soluble IL-6R? protein to mouse myeloma cells sp2/0. Ascites were used to produce the monoclonal antibodies (mAbs). The mAbs were purified by protein G immunoaffinity method. The mAb SI10 was used as coating antibody, the other mAb H126 recognized different epitope from SI10 was labeled by 125I. Results The immunoradio assay for detecting soluble IL-6R? was set up. It has high stability and accuracy. The detecting limit is 10 ng/ml. The serum concentration of soluble IL-6R? is (81.96 ? 7.23) ng/ml in healthy donors and (237.58?70.96) ng/ml in patients with multiple myeloma. Significant difference was founded between two groups (P
Résumé
Objective:To establish the sensitive,specific,stable and convenient sgp130 ELISA kit.Methods:The mAb T2 against human gp130 was used as coating antibody;the other mAb T12 recognized different epitope with T2 was labeled by biotin,then a ELISA kit for detecting sgp130 was set up.Results:sgp130 ELISA kit is successfully established and its sensitivity is 10 ng/ml.After the kit is placed in 4℃ for 3 months,the kit's CV is less than ?7.6% and the retrievable rate is 95%～111%.These indicate that it has highly sensitivity,stability and accuracy.The normal serum concentration of sgp130 in healthy donors is 536.92～287.88(ng/ml),but there is higher in patients with hyperthyroidism(937.16?217.5) and chronic nephritis(806.45?138.47).Significant difference is found comparing with normal control(P