Objective:To investigate the IRF5 rs2004640,rs10954213,rs4728142 loci gene polymorphisms in the progress of Systemic lupus erythematosus(SLE),and explore the influence of the IRF5 gene polymorphisms to the SLE.Methods:218 patients with SLE and 200 health controls were analyzed by using TaqMan-PCR.And all allele frequencies were calculated.The risk factors were compared between cases and controls.At the same time,antinuclear antibodies and double-stranded DNA antibody of 218 SLE cases were analysed with indirect immunefluorescence method,specific autoantibodies in plasma spectrometry were determined using linear immunoassay in plasma.Results:T allele frequency of IRF5 rs2004640 loci in SLE was higher than the controls,allele frequency of the G/T distribution differences in two groups were statistically significant(χ2=6.809,P=0.009).The GG/TT genotype frequencies between the control and the SLE were statistically significant(χ2=5.111,5.035;P=0.024,0.025).Compared control group with SLE group,G allele frequency of IRF5 rs10954213 loci in SLE group was higher than the controls,the difference was statistically significant(χ2=4.332,P=0.037).And GG genotype distribution had significant difference in the two groups(χ2=5.805,P=0.016).SLE group AA/AG/GG genotype distribution of IRF5 rs4728142 loci,compared with control group,there were no statistically difference(χ2=1.273,0.902,1.853;P=0.259,0.342,0.173).And allele frequency A/G also were not statistically differences in two groups(χ2=2.651,P=0.104).The T allele frequency of IRF5 rs2004640 and autoantibodies(anti-Sm,anti-Rib-P)significant associations were found with SLE patient.There were no statistically significant difference between IRF5 rs10954213 with autoantibodies.Compared with SLE remission patients,ANA,ds-DNA increased obviously.Anti-NUC,anti-His,anti-Rib-P were higher,there were statistically significant differences betwen two groups.Conclusion:The GG/GT/TT polymorphism of IRF5 rs2004640,GG/GA/AA polymorphism of rs10954213 were related to SLE.But the GG/GA/AA polymorphism of IRF5 rs4728142 were not significant difference.IRF5 rs2004640 T comtributed to the anti-Sm,anti-Rib-P.In active SLE patients,anti-ds-DNA,anti-NUC,anti-His,anti-Rib-P were higher than the other group.

Objective To investigate the correlation between single nucleotide polymorphism rs13146124 of interferon regu‐latory factor2(IRF2) on type Ⅰ interferon pathway and systemic lupus erythematosus(SLE) in a population from Guizhou Prov‐ince .Methods The polymorphism IRF2(rs13146124) was detected by using Taqman‐PCR in 366 cases of patients with SLE and 218 healthy controls .The genotype and allele frequencies were calculated and analyzed .Results The genotype frequencies of AA , AG and GG in IRF2 rs13142164 site in patients with SLE were 0 .011 ,0 .246 and 0 .743 respectively ,compared with the control group ,the difference was not statistically significant (χ2 =0 .093 ,0 .205 ,0 .136 ;P=0 .761 ,0 .651 ,0 .712) .The allele frequencies of A and G in IRF2 rs13146124 site in SLE patients were 0 .13 ,0 .87 respectively ,compared with the control group ,the difference was not statistically significant(χ2 =0 .071 ,P=0 .790) .There was no significant difference between the allele frequencies of A and G in IRF2 rs13146124 site in SLE and ANA ,dsDNA and other specific antibodies .There was no correlation between the allele frequen‐cies of A and G in IRF2 rs13146124 site in SLE and clinical features such as arthritis ,kidney damage ,etc .Conclusion The poly‐morphism of rs13146124 in IRF2 may not be associated with SLE in the population from Guizhou Province .

Objective:To investigate the chemokine receptors expression on regulatory T cells induced by different antigens and chemotaxis of T cells conducted by CCL20 and CCL22.Methods:BALB/c mice were divided into different groups and inoculated with skin-antigen derived from C57BL/6 mice or BCG vaccine respectively.The changes of CCR4 and CCR6 expression on CD4+CD25-T cells and CD4+CD25+CD127-T cells were detected at 1st,2nd,3rd and 4th week using flow cytometer.The chemotactic effects of CCL20 and CCL22 on CD4+CD25-T cells and CD4+CD25+CD127-T cells subsets were assayed by chemotaxis assay.Results: ①In skin-antigen group,the average fluorescence intensity ( MFI) of CCR4 on CD4+CD25-T cells at 4 week was significantly stronger than that at 3 week (P<0.05).There were no significant changes of CCR4 expression in BCG group.②The MFI of CCR6 on CD4+CD25-T cells was strongest at 2 week in skin-antigen group (P<0.05) while in other groups at 4th week (P<0.05).Besides,the expression of CCR6 on CD4+CD25+CD127-T cells was stronger during the first two weeks than the later two weeks ( P<0.05) in skin-antigen group, while in BCG group,the MFI was stronger at 2nd and 4th week than 1st week (P<0.05).③The chemotactic index of CD4+CD25+CD127-T cells was highest at 4th week in BCG group (P<0.05) in CCL20 induced chemotaxis,while in other groups were higher at 2nd and 4th week(P<0.05).In CCL22 induced chemotaxis ,there were no significantly differences of chemotactic index of CD4+CD25+CD127-T cells between skin-antigen group and BCG group.Conclusion:①The expression of chemokine receptor on the surface of Treg was associated with antigenic properties.②CCL22 had a notable chemotactic effect on Treg at the early stage of post-induction, while CCL20 did that at the late stage of post-induction.

Objective:To explore the expression and location of TLR5 and NLRC4 on different breast cancer cell lines MDA-MB-231,MCF-7 and MDA-MB-435 and TLR5 activation in breast cancer cell line by recombinant flagellin . Methods:The mRNA level of TLR5 and NLRC4 in MDA-MB-231, MCF-7 and MDA-MB-435 cell were detected with quantitative Real-time PCR and TLR5 expression and location in MDA-MB-231 and MCF-7 cell were detected with Flow cytometry. Induction,expression,purification and i-dentification of recombiant flagellin,including FliC (activating both TLR5 and NLRC4),FliC△90-97(unable to activate TLR5),FliC-L3A (unable to activate NLRC4),FliC△90-97:L3A (unable to activate both TLR5 and NLRC4). 1 μg/ml recombinant flagellin were used to stimulate MCF-7 cell lines,12 h later,the supernate were collected,and ELISA was performed to assess the secretion of IL-8. Results:The mRNA level of TLR5 in MCF-7 cell was 1 700 folds higher than that of MDA-MB-435. TLR5 was expressed in MCF-7 cell surface and ctyosol,while expressed only in cytosol in MDA-MB-231 cell. FliC and FliC-L3A,which were able to activate TLR5 pathway,stimualted MCF-7 cell line to secret IL-8,but FliC△90-97 and FliC△90-97:L3A did not. Conclusion:TLR5 and NLRC4 have been expressed in different breast cancer lines,but there exists difference on the expression level and location of TLR5. Expression level of TLR5 and NLRC4 in MCF-7 cell were higher than other breast cancer lines. TLR5 receptor which is expressed on the surface of breast cancer cell can be activated by flagellin,and these work also provide us experimental basis to further understand the impact of TLR5 activation on breast cancer cell proliferation.

Objective:To study the effect of chemokines CCL20 and CCL22 combined with skin-induced Treg on survival time of grafted skin.Methods: Skin grafting mice were divided into four groups, three mice per group, namely Treg group, Treg+CCL20 group,Treg+CCL22 group and control group.C57BL/6 mice were used as donor and BALB/c as acceptor, and the Treg cells were isolated from the mice induced by skin allograft.After skin grafted,CCL20 and CCL22 were subcutaneous injection every day,which lasted for 10 day.Survival time of skin in each group were observed and recorded.The Treg colonzation experiments were performed as follows.We firstly isolated Treg with Magnetic cell sorting system( MACS) and then labled them with 99 Tcm.After that we intravenously injected them into the mice.3 hours later,the mice were sacrifced and the radioactivity of organs were detected by GC-2016γradioim-munoassay counter.Results:①After Treg treated the survival time of skin grafted in antigen-induced Treg group was signifiantly longer than control group,when treg were cooperated with CCL20 and CCL22,the skin grafted showed more longer survival time than Treg and control groups( P<0.001 ).②After injection of induced Treg, Treg in autologous and allogeneic skin grafts goups were mainly distributed in autologous and allogeneic skin,accounting for 60% and 98% respectively.When cooperated with CCL20 or CCL22,the Treg were mainly distributed in liver.Conclusion:Chemokines CCL20 and CCL22 synergistically improved the effects of skin antigen induced Treg on survival time of skin graft,which probably related with the Treg colonization into the liver.

0.05).Compared to cultivation whose positive rate was 33.33%,the rates were obvious higher(P