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Objective To explore the relationship between HBV infection and the genotypes and allele frequencies of CⅡTA G-944C gene polymorphism in three minority populations(Jinuo,Dai and Aini population)in Xishuangbanna district,Yunnan province.Methods Polymerase chain reaction and sequencing method were used to study the genotypes and allele frequencies distributions of CⅡTA G-944C gene polymorphism in those three populations.Relationship between the genotypes distribution and HBV infection results were also analyzed.Results The rates on HBV infection and HBsAg carrier status in Aini minority population were 89.2% and 16.3%,which were significantly higher than in Jinuo(27.9% and 3.9%,χ~2=135.196 and 10.361,P=0.000 and 0.001)and Dai population(44.9% and 6.6% χ~2=96.783 and 8.748,P=0.000 and 0.003)while among Aini population it was significantly different with the other two minority populations.The CC genotype and C allele frequencies were more distributed in Aini population than in the other two minority populations.In contrast,the GG genotype and G allele frequencies were lower than the other two minority populations,with χ~2 rates between Aini and Jinuo population were 11.841 and 12.208 and the P as 0.003 and 0.000 respectively while the χ~2 rates between Ami and Dai population were 23.902 and 20.220 with P value as 0.000 and 0.000.The genotypes frequencies of CⅡTA G-944C was significantly different in the infected individuals(IF)group and health control(HC)group in Jinuo population(χ~2=6.150 and 4.911,P=0.046 and 0.027).When compared with HBsAg+ group and HBsAg~- group,the genotypes and allele frequencies were different in Aini population and the total three minority populations(χ~2 rates in Jinuo minority were 8.650 and 5.034 with P values as 0.013 and 0.025).However,the χ~2 rates in the whole population were 13.047 and 9.416 with P values as 0.001 and 0.002,respectively.The distribution of CC genotype and C allele gene in HBsAg~+ group was increasing.Data from non-condition logistic regression analysis and adjusting for confounding factors,the HBsAg~+ group had a significantly increase of HBsAg~- group under the C allele Recessive Model(P=0.000;OR=2.964;95% CI:1.609-5.460).Conclusion The genotypes and allele frequencies distribution of CⅡTA G-944C were different in the three ethnic populations.Polymorphism of this gene was closely associated with HBsAg carrier.The CC genotype patients were more easily to become HBsAg carrier.
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Objective:To Construct the eukaryotic expression vectors containing four different haplotypes DNA of human CIITA gene promoter IV.Methods:Four haplotypes(CG,CC,TG and TC)can be constructed based on two single nucleotide polymorphism(SNPs)sites(G-944C and T-1350C)in promoter IV of human CIITA gene.The 487bp DNA fragments of CIITA promoter IV including the two SNPs were obtained by polymerase chain reaction(PCR)based on human genome DNA from the subjects with the CG/CG,CC/CC,TG/TG and TC/TC genotypes,which were TA cloned to pMD18-T Simple vectors and then were digested by restriction endonucleases Mlu I and Hind Ⅲ.After fragment recovery,those were ligated to four PGL3-Basic Vectors and four PGL3-Promoter Vectors respectively,which were digested by restriction endonucleases Mlu I and Hind Ⅲ as well.All recombinant plasmids were identified by sequencing.Results:Eight recombinant plasmids(CG-PGL3-Basic,CG-PGL3-Promoter,CC-PGL3-Basic,CC-PGL3-Promoter,TG-PGL3-Basic,TG-PGL3-Promoter,TC-PGL3-Basic,TC-PGL3-Promoter)containing four different haplotypes DNA of human CIITA promoter IV were obtained,whose sequences completely matched with the theoretical prediction demonstrated by sequencing.Conclusions:The eukaryotic expression vectors containing four different haplotypes DNA of human CIITA promoter IV are successfully constructed,and it lays the groundwork for the further study of different haplotype function.
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Objective To investigate the relation and difference of expression phase between classⅡtransactivator (CⅡTA) and HLA-DR antigens after IFN-? incubation, so as to investigate the potential effect of the class Ⅱ trasactivator (CⅡTA) in graft-versus-host disease(GVHD). Methods T lymphocyte from peripheral blood of health was incubated with IFN-? for 1 000 U/ml. RT-PCR was used to detect CⅡTA mRNA and Western blot was used to explore HLA-DR antigen in various time periods. Then Stat1? antisense oligonucleotides (AS) were given to inhibit the expression of CⅡTA, CⅡTA mRNA and HLA-DR antigen were tested again at peak point. Results CⅡTA mRNA was detectable 5 h after IFN-? treatment and peaks at 14 h; HLA-DR protein was detectable 28 h after IFN-? treatment and peaks at 52 h. The expression of CⅡTA mRNA and HLA-DR protein was lower in AS groups than that in control groups. Conclusion CⅡTA expression was positively correlated to HLA-DR expression, and earlier than the HLA-DR expression after IFN-? incubation. CⅡTA might be used as an early predicting marker of GVHD.
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Objective To investigate the role of MHC class Ⅱ transactivator(CⅡTA) in upregulating the expression of HLA class Ⅱ antigen in HepG2 cells.Methods Eukaryotic expression vector EBS-NPL-CⅡTA containing CⅡTA cDNA or the empty vector EBS-NPL was transfected into HepG2 cells respectively.CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were respectively detecued by RT-PCR,indirect cell immunofluorescence technique and flow cytometry in original HepG2 cells,HepG2 cells transfected with EBS-NPL-CⅡTA or the empty vector EBS-NPL.Results The expression of CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were not observed in original HepG2 cells and HepG2 cells transfected with empty vector,but in the HepG2 cells transfected with EBS-NPL-CⅡTA.Conclusion CⅡTA is a switching factor of mastering the expression of HLA class Ⅱ antigen in HepG2 cells.The lack of CⅡTA expression in HepG2 cells contributes to no expression of HLAⅡ antigen.
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0.05). Conclusion The polymorphism of G-944C in CⅡTA gene promoter Ⅳ was associated with the susceptivity of chronic HBV infection, but was not associated with severity of diseases. The individuals with chronic HBV infection of CC genotype are of less possibility to develop chronic liver disease than those of other genotypes.