Preliminary Sieving Of Anti-megakaryocyte Autoantibody Ralated Antigen In Systemic Lupus Erythematosus With Thrombocytopenia

Objective: Preliminary sieving of anti-megakaryocyte autoantibody ralated antigen in systemic lupus erythematosus associated with thrombocytopenia.Methods: Collect the serums of systemic lupus erythematosus pateints with thrombocytopenia and health group,sieving of megakaryocyte antibody positive serums by Indirect immunofluorescence method. Total proteins in MEG-01 cells that cultured in vitro were extracted and separated by 2-DE.The separated protein bands were then transfered to polyvinylidene fluoride membrane. The anti-megakaryocyte antibody positive serum of systemic lupus erythematosus patients with thrombocytopenia as the experimental group and the serum of health people as the control grop,incubated with serums of healthy people and patients as the source of primary antibody for western blotting.Use the ImageMasterTM2 D Platinum image analysis software to analyze the hybrid proteins spots of each group.Results: 1.The results of Indirect immunofluorescence:8 patients have been detected anti-megakaryocyte antibody positive of 35 cases of systemic lupus erythematosus patients with thrombocytopenia,positive rate is 22.9%,20 cases of healthy control group have not been detected anti-megakaryocyte antibody, positive rate is 0%. 2.Total proteins in MEG-01 cells were extracted and separated by two-dimensional-electrophoresis(2-DE) and dyed by coomassie brilliant blue,after ImageMasterTM2 D Platinum image analysis software analyze,a total of 545±27 protein spots were detected.3.The results of Two-dimensional- electrophoresis combine with western blotting: analysis through ImageMasterTM2 D Platinum image software, a total of 2 disparate protein spots were found between systemic lupus erythematosus patients with thrombocytopenia whose anti-megakaryocyte antibody are positive and health grop whose anti-megakaryocyte antibody are negative, the 2 protein spots molecular weight are about 48 kD.Conclusions:1.Systemic lupus erythematosus patients with thrombocytopenia have been detected anti-megakaryocyte antibody, regular health group have not been detected the antibody of megakaryocyte. 2.There are many antibodies in the serum of systemic lupus erythematosus patients with thrombocytopenia, two of them are related with thrombocytopenia,the 2 protein spots molecular weight are about 48 kD.