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OBJECTIVE@#To explore the relationship between the level of soluble HLA-E (sHLA-E) molecules in plasma and gene polymorphism and leukemia in Shenzhen of China.@*METHODS@#Enzyme-linked immunosorbent assay was used to detect sHLA-E level in plasma of 103 leukemia patients and 113 healthy blood donors. PCR-SBT was used to identify the HLA-E genotype of 73 leukemia patients and 76 healthy blood donors.@*RESULTS@#The level of plasma sHLA-E of 103 leukemia patients was significantly higher than that of 113 healthy blood donors (P<0.001); And the level of plasma sHLA-E in 77 myeloid leukemia patients was also significantly higher (P<0.001). The percentage of patients with plasma sHLA-E concentration of 0-199 ng/ml in leukemia and myeloid leukemia patients was 37.86% and 32.47%, respectively, which was significantly lower than 53.98% of healthy donors, the difference was statistically significant (P<0.05, P<0.01); While, when the plasma sHLA-E concentration was more than 400 ng/ml, the percentage was 33.01% and 36.36%, respectively, which was significantly higher than 13.28% of healthy donors, the difference was also statistically significant (P=0.001, P<0.001). There was no significant difference in the level of plasma sHLA-E among different HLA-E genotypes (P>0.05), whether healthy blood donors or leukemia patients.@*CONCLUSION@#The level of plasma sHLA-E in patients with leukemia (especially myeloid leukemia) is significantly higher than that of healthy blood donors, but different HLA-E genotypes do not affect the level of plasma sHLA-E. A cut-off value for the concentration of plasma sHLA-E (recommended risk value >400 ng/ml) can be set to assess the risk of certain pre-leukemia patients.
Subject(s)
Humans , Genotype , Histocompatibility Antigens Class I/genetics , Leukemia/genetics , Polymorphism, GeneticABSTRACT
ObjectiveTo explore the mechanism of Si Junzitang in regulating the expression of NKG2A to affect the anti-colon cancer function of natural killer （NK） cells. MethodNK cells isolated from healthy honors were cultured and used to construct the three incubation models of NK cells， human colon cancer HCT116 cells， and NK cells + HCT116 cells （co-incubation）. real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was conducted to determine the mRNA levels of natural killer group 2 member A （NKG2A） and interleukin （IL）-15 in NK cells， as well as the mRNA level of histocompatibility leucocyte antigen E （HLA-E） in HCT116 cells. The secretion of IL-15 was detected by enzyme-linked immunosorbent assay （ELISA）. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the applicable concentration of IL-15 and test the effects of Si Junzitang and IL-15 on the activities of NK cells and the HCT116 cells in the co-incubation model. The effects of Si Junzitang and IL-15 on the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells were detected by Real-time PCR. Monalizumab （M， anti-NKG2A mab） was used to block the NKG2A-HLA-E pathway in co-incubation model， and then the proliferation of HCT116 cells was detected by MTT assay. ResultThe interaction of NK cells and HCT116 cells up-regulated the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells （P<0.05）， as well as the expression level and secretion of IL-15 （P<0.05）. Compared with the blank group， Si Junzitang and Si Junzitang + IL-15 promoted the proliferation and improved the anti-colon cancer function of NK cells （P<0.01）. Furthermore， they down-regulated the mRNA levels of NKG2A in NK cells and HLA-E in the HCT116 cells co-incubated with NK cells （P<0.01）. M and IL-15 + M inhibited the proliferation of HCT116 cells compared with the groups without M （P<0.01）. ConclusionThe interaction of NK cells and HCT116 cells can induce activation of NKG2A-HLA-E pathway to impair NK cell function. Si Junzitang can inhibit the activation of NKG2A-HLA-E pathway to restore the anti-colon cancer function of NK cells.
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Entre mulheres com HIV/AIDS há um maior número de casos de infecções persistentes pelo HPV contribuindo para um risco aumentado do desenvolvimento de lesões intraepiteliais escamosas cervicais. Ademais, a expressão anormal de moléculas HLA-E pode modular o sistema imunológico através da ligação com o receptor inibitório (CD94/NKG2A) ou estimulatório (CD94/NKG2C) de células NK e linfóticos T CD8+, diminuindo imunovigilância favorecendo a evasão de céulas infectadas por vírus. Diante da escassez de estudos avaliando a molécula HLA-E na interação com o HPV, mais especificamente na infecção pelo HIV-1, este estudo teve como objetivo avaliar a expressão de HLA-E em lesões intraepiteliais cervicais em mulheres portadoras do HPV, apresentando ou não a infecção pelo HIV-1. Trata-se de um estudo transversal, ao qual foram submetidos ao processo imunohistoquímico tecido do colo do útero parafinado de 67 mulheres infectadas pelo HIV-1 e 62 mulheres não infectadas, todas com lesão intraepitelial cervical com HPV, o qual foi tipificado. A expressão da molécula HLA-E foi analisada quantitativamente como sem expressão, de 1% a 30%, de 31% a 70% e de 71% a 100%. Os resultados mostraram que a infecção por herpes vírus foi maior entre as participantes HIV+ (P=0,005). Na análise imunohistoquímica, ficou evidente que as lesões intraepiteliais cervicais de mulheres infectadas pelo HIV-1 apresentaram redução na expressão da molécula HLA-E de 31% a 100% em comparação com mulheres sem a infecção pelo HIV- 1 (P=0,001), sugerindo que essa redução possa ser um mecanismo de escape viral, que acarreta a redução da apresentação de peptídeos virais para os linfócitos T CD8+. A expressão do HLA-E não foi associada aos graus de lesões intraepiteliais cervicais. Outros estudos são necessários para melhor compreensão do padrão e da função da expressão das moléculas HLA-E em lesões intraepiteliais cervicais de mulheres infectadas pelo HIV- 1
Among women with HIV / AIDS there is a greater number of cases of persistent HPV infections contributing to an increased risk of developing squamous intraepithelial lesions of the cervix. In addition, abnormal expression of HLA-E molecules can modulate the immune system by binding to the inhibitory (CD94 / NKG2A) or stimulatory (CD94 / NKG2C) receptor NK cells and CD8+ T lymphocytes, decreasing immunovigilance and favoring the evasion of infected cells infected with virus. Due to the lack of studies evaluating the HLA-E molecule in the interaction with HPV, more specifically in HIV-1 infection, this project aimed to evaluate the expression of HLA-E in cervical intraepithelial lesions of infected women or not by HIV- 1, with the infection by HPV. It is a cross-sectional study, which was submitted to immunohistochemical processing the paraffin-embedded cervix tissue of 67 HIV-1 women infected with HIV-1 and 62 uninfected women, all of them with cervical intraepithelial lesion with HPV, which was typified. The expression of HLA-E was quantitatively analyzed as non- expressed, 1% to 30%, 31% to 70% and 71% to 100%. The results showed that the herpes virus infection was higher among HIV + participants (P = 0.005). In immunohistochemical analysis, it became evident that cervical intraepithelial lesions in HIV-1 infected women showed a reduction the expression of HLA-E molecule from 31% to 100% compared to women without HIV-1 infection (P = 0.001) Suggesting that this reduction may be a viral escape mechanism, which leads to a reduction in the presentation of viral peptides to CD8+ T lymphocytes. The expression of HLA-E was not associated with cervical intraepithelial lesions. More studies are need to better understand the pattern and function of HLA-E molecule expression in cervical intraepithelial lesions of infected women by HIV-1
Subject(s)
Humans , Female , Cross-Sectional Studies , HIV , Squamous Intraepithelial Lesions of the Cervix , PapillomaviridaeABSTRACT
Aim According to various target sites of HLA-E mRNA,to design and synthesize 3 pieces of HLA-E siRNA chain,to compare quantitatively their efficiency of silencing gene in BEL-7402 with HLA-E(+)in order to select the dominant siRNA.Methods The hepatocarcinomal BEL-7402 cells,induced by 5×10~5 IU·L~(-1) IFN-γ,expressed HLA-E(+) and was pured by flow cytometry selecting as target cells for research.3 pieces of specific siRNA(A,B,C group)were designed and chemically synthesized,then the concentration of which(0.1 mmol·L~(-1))was respectively transfected through Lipofectamin 2000 into target cells.After 48 h,the gene silent effect on HLA-E gene in A,B and C groups was quantitatively observed by cytoimmunofluorence,flow cytometery,Western blot and real-time PCR,as well as on NK cytotoxicity to target cells tested by NK killing rate.Results Compared with those of control or non-specific siRNA group,HLA-E antigen,protein product,HLA-E mRNA and HLA-E molecule on cell surface were statistically down-regulated in A,B,and C group(P<0.01),whose were silenced more (above 90%) in B or C group than in A group (P<0.01).The NK killing rate in A,B and C groups was dominantly improved(P<0.01),which in B or C group was higher than in A group (P<0.01).Conclusion The targeted siRNA can specifically and high-efficiently silence HLA-E expression in hepatocarcinomal cells,and may keep them from immunoescape through non-classic HLAⅠ pathway to imply new strategy for hepatocarcinomal gene-immunotherapy.
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Objective:To explore the mechanism of association between recurrent spontaneous abortion(RSA) and the polymorphism of HLA E,had detected the frequencies of five HLA E alleles in 53 women with RSA and 156 random Shanghai Chinese controls.Methods:PCR sequence specific oligonucleotide (PCR SSO) typing was carried to detect polymorphism in exon2 and exon3 of HLA E gene.Results:HLA E *0101 is the most common allele in both RSA patients and controls(50% and 43.09% respectively).HLA E *01031 and E *01032 were detected with allele freqencies of 23.79% and 33.12% in controls,20.75% and 29.25% in RSA.No HLA E *0102 and E *0104 alleles could be detected in both groups.Conclusion:Maternal HLA E polymorphism does not associated with pathogenesis of RSA.
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Objective:To explore the influence of chaperone tapasin on HLA E expression on cell surface.Methods:The infection method was used to transfer the recombinant retrovirus expression vector which was constructed with HLA E molecules on target cells was detected with FACS technique.Results:Exogenous HLA E expression on T2 cell surface was detected with 74.13% and no HLA E expression was detected on control cells and that was transfected with empty vector.Conclusion:It is possible that the expression of HLA E molecules on cell surface involves in TAP independent mechanism.