Serum Proteomicsresearch On Normal Pregnancyand Active Immunotherapyof Recurent Spontaneous Abortion

Objective:This study applied proteomics method to select the the serum discrepant proteome of normal pregnant women and the patients before and after the active immunotherapy who with recurrent spontaneous abortion (RSA). The objective was searching for the possible immune tolerance of pregnancy regulatory protein, and preliminary investigating the possible serological pathogenesis of RSA and the possible humoral immune efficacy mechanism of active immunotherapy.Method:Selected a total of 25 samples from patients with RSA who are negative of blocking antibodies(BA) before and after the active immunotherapy as the self-control patient groups. In addition selected a total of 20 samples from initial normal pregnant women in the same period as the normal control group. Estimated the BA of the three groups respectively with ELISA, and estimated the serum proteome of the three groups with SELDI-TOF-MS and WCX-2. Selected the significant different serum proteins and established two diagnosis tree-models. In further screened of the possible pregnancy immunological tolerance regulatory protein by comparing differences proteins in the three groups.Results:The positive rate of BA in normal pregnant women was only 10%. The positive rate of BA in patients with RSA was 11.1%. The positive rate of BA in RSA patients who were negative changed to 28.0% after one course of active immunotherapy. Estimated the serum proteomics, it found that there were in total of 85 protein peaks with statistical difference (P<0.05) and in total of 29 protein peaks with significant statistical difference(P<0.01) between the RSA before immunotherapy group and normal control group. It developed two protein peaks (M/Z is 13758.0,3450.94)could separate the patients with RSA and normal pregnancy women. The distinguish had sensitivity of 100% and specificity of 100%. There were in total of 37 protein peaks with statistical difference (P<0.05) and in total of 10 protein peaks with significant statistical difference (P<0.01) between the RSA before immunotherapy group and after immunotherapy group. It developed two protein peaks (M/Z is 13758.0,3450.94)could separate the RSA patients who is before immunotherapy and after immunotherapy. The distinguish had sensitivity of 100% and specificity of 88%. In these proteins of both had statistical difference(P<0.05) between normal contorl group and RSA before immunotherapy group, and between RSA before immunotherapy group and RSA before immunotherapy group, there were in total of 17 protein peaks both expressed higher or lower in normal control group and RSA after immunotherapy group than RSA before immunotherapy group.Conclusion:BA may not be the key factors in the normal immune tolerance. BA may not suitable as a triage criteria for RSA patients whether receive active immunotherapy or not and may not as a specific indicator for assessing the effect of immunotherapy either. The protein mass spectrometrys are express discrepantly between normal contorl group and RSA before immunotherapy group, and between RSA before immunotherapy and after immunotherapy. some discrepant proteins express consistent trend both in RSA after immunotherapy group and normal contorl group, while express reverse trend in RSA before immunotherapy group. It prompt that the discrepant proteins very likely not only play an important role in the mechanism of immune tolerance of normal pregnancy, but also in the pathogenesis of RSA and the efficacy mechanism of active immunotherapy.