Changes Of Cytokines In Immune Thrombocytopenic Purpura By Protein Array Detection

Background and objective:Immune thrombocytopenic purpura is the most common hemorrhagic disease. The incidence rate is 1.6-10/100,000. Although ITP is a benign disease, it usually relapses and progress into refractory ITP. A research on health related quality of life(HRQOL) in USA shows that the quality of life of ITP patients is beneath not only the common people but patients of other chronic diseases such as hypertension and arthritis.Despite of multiple therapies,ITP has no radical method yet.The pathogenesis remains unknown yet.The classical mechanism is that platelets sensitized by autoantibody are over destroyed through mononuclear-macrophage system. Recent researches find out that cellular immunity is also important in the pathogenesis besides humoral immunity.The in-depth study in the autoimmunity of pathogenesis of ITP has lead many new oriented immunologic interventions into clinical research. Protein array is a kind of high throughput monitoring system. It monitors the protein-protein interaction through the target molecules and capturing molecules interaction.Protein array can detect varieties of cytokines, which does help to clarify the pathogenesis of ITP and provide a theoretical basis for the targeted therapy.Objective:To detect the changes of 507 kinds of cytokines in ITP patients, screen out the cytokines possibly related to the pathogenesis of ITP, and then observe their changes in the signaling pathways by cluster analysis.Method:(1)Patients and controls:During August 2009 to April 2010,17 ITP patients were enrolled by the diagnostic criteria for ITP, including 12 primary,2 CR,3 relapsed patients.The primary patients'platelet counts range from 1-82*10-9/L,the mean count is 25*10-9/L. The relapsed patients'platelet counts range from 6-41*10-9/L,the mean count is 17*109/L. The relapsed patients'platelet counts range from 128-154*109/L. Patients complicated with viral hepatitis,diabetes,hypertension,cardiovascular diseases,pregnancy, active infection,or connective tissue diseases were excluded. The control group consisted of 7 adult healthy volunteers.Their platelet counts range from 128-263*10-9/L.(2) Detection of changes of 507 kinds of cytokines in ITP patients by RayBio(?) Biotin Label-based Human Antibody Array I.Results:It was found that 400 kinds of cytokines have changed, of which 110 elevated,175 decreased, while 115 elevated or decreased during the different stages of the disease. Compared with the healthy volunteers, the primary ITP patients showed 35 elevated and 130 decreased cytokines, the relapsed patients showed 99 elevated and 189 decreased cytokines,the CR patients showed 94 elevated and 144 decreased cytokines.Through the pathway analysis,we discovered 6 interested signaling pathways:cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, TGF-beta signaling pathway,chemokine signaling pathway,MAPK signaling pathway, TLR signaling pathway.Conclusion:Protein array,is an ideal method to screen the cytokines in ITP with little sample volume. The change in the cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, TGF-beta signaling pathway,chemokine signaling pathway,MAPK signaling pathway and TLR signaling pathway we observed maybe related to the pathogenesis in ITP.