Single Nucleotide Polymorphisms Of TNF-α Promoter And IFN-γAssociate With Susceptibility Of Immune Thrombocytopenia In Chinese Han Adults

Part Ⅰ The association between TNF-α promoter single nucleotide polymorphisms and susceptibility of immune thrombocytopenia in Chinese Han adultsObjectives:This study attempts to investigate the association between five single nucleotide polymorphisms (SNPs)(-238,-308,-857,-863and-1031) spanning the TNF-α gene promoter and the susceptibility of primary immune thrombocytopenia (ITP) in Chinese Han adults.Methods:In this study,215adult primary ITP patients and206healthy controls were included. Five polymorphisms in the TNF-α gene promoter were detected by PCR-RFLP and PCR-SSP. The χ2test or fisher’s exact test was used to compare frequencies of genotypes and alleles between patients and controls. Haplotypes were analyzed with the SHEsis online program.Results:None of the five TNF-α polymorphisms tested individually was significantly associated with the susceptibility of primary ITP patients with the acute form in Chinese Han adults. Whereas the haplotype GGC of TNF-α promoter SNPs (-238G/-308G/-857C) was significantly associated with a decreased susceptibility in controls and patients with this haplotype, especially in the male subgroup (P<0.05, OR=0.64,95%CI:0.42-0.97). Conclusions:This study represents the first report in Chinese Han adults that the single TNF-a loci haplotype GGC (-238G/-308G/-857C) can be differentially associated with the susceptibility of primary ITP. However, we failed to find the association between individual sites-238,-308,-857,-863and-1031in TNF-a promoter and the susceptibility of adult primary ITP. Part II The association between IFN-y single nucleotide polymorphism and susceptibility of immune thrombocytopenia in Chinese Han adultsObjectives:Interferon-gamma (IFN-y) participates as a strong candidate susceptibility factor for immune thrombocytopenia (ITP). This study attempts to investigate the association between the single nucleotide polymorphisms (SNPs) in+874site of the IFN-y gene and the susceptibility of primary ITP in Chinese Han adults.Methods:In this study,82adult primary ITP patients and79healthy controls were included. The IFN-γ+874polymorphism of the IFN-y gene was detected by PCR-SSP. The χ2test test was used to compare frequencies of genotypes and alleles between patients and controls.Results:The IFN-γ+874polymorphism tested was not significantly associated with the susceptibility of primary ITP patients with the acute form in Chinese Han adults.Conclusions:There was no association found between the IFN-γ+874polymorphism and the susceptibility primary ITP in Chinese Han adults. Part Ⅲ The expression of TNF-α, IFN-γ and other associated genes in peripheral blood mononuclear cells of adult patients with primary ITP.Objectives:This study attempts to examine the expression of T cell subset associated genes and discover the immunal mechanisms and evaluate the therapy effect of primary ITP.Methods:We detected TNF-α、IFN-γ、TIM-3、Gal-9、FOXP3and IL-17in PBMCs of37preliminary diagnosis and therapy finished samples of primary ITP patients and20healthy controls by SYBR Green Ⅰ qRT-PCR. Meanwhile, we analyzed the correlation between the expression and primary ITP by statistical analysis.Results:The mRNA relative expression levels of IFN-y (P<0.001), TIM-3(P<0.05), Gal-9(P<0.01), IL-17A(P<0.001) in PBMCs of preliminary diagnostic primary ITP adults were significant higher than healthy controls. The relative expression levels of IFN-y(P<0.01), TIM-3(P<0.05), Gal-9(P<0.01) and FOXP3(P<0.001) in peripheral PBMCs of primary ITP adults treated with corticosteroids in the remission state were significant lower than those in the preliminary diagnostic state.Conclusion:The expression levels of IFN-y, TIM-3, Gal-9, IL-17A and FOXP3were found significantly associated with adult primary ITP during the onset and treatment process. The study of these molecular and correlated cells would contribute to the diagnosis and treatment of primary ITP and the discoverary of the mechanisms in this disease.