Expression and significance of MICA, NF-κB and P53 in carcinoma of the urinary bladder / 中国医师杂志

ObjectiveTo investigate the mechanism of the expression of MHC class Ⅰ chain-related A (MICA) in carcinoma of the urinary bladder,the relationship between MICA,nuclear factor-κB ( NF-κB) and p53 expression in bladder cancer was studied.MethodsThe expression of MICA,NF-κB and p53 in a total of 75 cases of urothelial carcinoma tissues and 15 normal mucous membrane tissues of the bladder was evaluated by immunohistochemistry.ResultsThe expression rates of MICA,NF-κB and p53 protein in urothelial carcinoma were 4.08％,85.3％ and 49.3％,respectively.Up-regulation of their expression was found in urothelial carcinoma compared with normal mucous membrane tissues ( P ＜ 0.05 ).MICA expression was positively correlated with NF-κB expression( r =0.256,P =0.027),but negatively correlated with p53 expression( r =- 0.23,P =0.047 ).ConclusionsUp-regulation of MICA expression was detected in bladder cancer.MICA protein may be a new tumor-associated antigen of bladder cancer.MICA expression may be regulated by NF-κB pathway,while p53 pathway may not play a part in MICA up-expression during the urothelial malignant transformation.

Determination of antigen-specific CTL induced by the gene vaccine in vitro with MHC pentamer / 中华检验医学杂志

Objective To determine the antigen-specific CTL in PBMC induced by a fusional family-gene vaccine of the immunoglobulin heavy chain variable gene framework region combined with the sequence of cytokine CM-CSF in vitro with MHC pentamer. Methods Peripheral blood samples were collected from two healthy donors and two patients. One was follicular lymphoma and another was hair cell leukemia. PBMC were isolated by density gradient centrifugalization with Ficoll and then subsequently differentiated into immature DCs (imDCs) induced by recombinant human GM-CSF and recombinant human IL-4. Gene gun was used to deliver the plasmids of the gene vaccine or the control plasmids into the imDCs. RT-PCR and ELISA assay were used to detect IgHVl-GM-CSF mRNA and GM-CSF in order to validate the transfection of the vaccine. After adding the cytokine cocktail, the imDCs became mature DCs. Then the mature DCs were co-cultured with lymphocytes from the blood samples for the induction of the antigen-specific CTL. The cultured cells were classified into vaccine group and control group and harvested at different time points of 0 d,7d, 17 d and 24 d after transfection. The subset of CD3+CD8+ T cells was analyzed by FCM assay. Finally, the CTL levels were detected with fluorescently labeled MHC pentamer antibody targeting vaccine epitopes. Results With the induction of cytokines, the imDCs with typical morphology were generated in PBMC. After delivering, the efficient expressions of the vaccine in the imDCs were determined by RT-PCR. And ELISA results also confirmed that GM-CSF was produced at a level of (28 ±6) ng/106 cells of the imDCs loaded with the vaccine, which was significantly different from that of control group (10 ± 3) ng/106 cells (t = 5. 191, P <0.01). FCM assay result showed that the CD3+ CD8+ T cells increased in a stepwise pattern during the culture. For control group, the levels at 0 d,7d, 17d and 24 d were ( 34. 24 ± 2. 72 )% , (46.06 ± 3.08)%, ( 65. 34 ± 4. 26 )% and (73.86 ±4.85 )% , respectively. For vaccine group, the results were (32. 28 ± 2. 08 ) % , (45. 32 ± 3. 81)% , ( 63. 37 ± 4. 21)% and (75. 01 ±3. 20)%. The differences between each time point had statistical significance (F = 176. 966 ,P <0.01) ,but there was no statistical differences between vaccine group and control group ( F = 0.657,P>0.05). The MHC pentamer analysis showed that the DCs loaded with IgHV1-GM-CSF fusional vaccine could efficiently induce the antigen-specific CTL response and the CTL levels increased gradually with the culture time, with the highest level of 4. 36% in the lymphoma blood and 3. 89% in the hair cell leukemia blood. Conclusions MHC pentamer assay could efficiently determine the antigen-specific CTLs response induced by the gene vaccine of family IgHV frame region in vitro. It could be a useful method for monitoring of anti-tumor cell immunity and evaluating of diagnosis and prognosis of the tumors in clinical application.

Effects of human leukocyte antigen-G on proliferation and invasion of JEG-3 cell line / 中华妇产科杂志

Objective To investigate the effect of human leukocyte antigen-G(HLA-c)on the growth and invasion of JEG-3 cell line and the role of HLA-G in the onset and development of pre-eclampsia.Methods The experiment was composed of three groups:groups of transfection,negative control and blank control.which corresponded to groups of HLA.G siRNA transfection,negative siRNA transfection and no transfection HLA-G overexpressed choriocarcinoma cell line JEG-3 was used.The role of HLA-G in JEG-3cell monolayer was examined by RNA interference technology using HLA-G specific small interfering RNA (siRNA).Expression of HLA.G was detected by reverse transeriptase-polymerase chain reaction and western blot analysis.Changes of cell cycle,apoptosis,proliferation and invasion were respectively detected by methvl thiazolyl tetrazolium(Ma r).flow cytometry assay and transwell test.Results (1)The mRNA and protein levels of HLA.G control group and blank control group were 0.0013±0.0014.0.0163 ±0.0007 and 0.1923 ±0.0384.0.2184 ±0.0153,respectively,which were both significantly different(P<0.05);the number of negative transfcction group was 0.1606±0.0133 and 0.2020±0.0132.which had no significant difference compared with blank control group(P>0.05).(2)The integral absorbance(IA)valUCB of the HLA-G transfecfion group and blank control group were 0.44±0.04 and 0.75±0.13 respectively.which was significantly different(P<0.01);the/A value of negative control group was 0.69±0.10.which was not significantly different compared with blank group(P>0.05).(3)The ratios of G2/M and S phase cells in transfection group were(10.9±2.2)%and(58.6±0.8)%respectively,significantly different compared with the blank control group[(15.4±1.9)%and(52.9±2.3)%respectively;P<0.01].(4)The ratio of early apoptosis cells in transfection group[(14.5±2.7)%]Was significantly increased compared with neg~ive[(5.3 ±1.1)%]and blank control group[(4.7±0.6)%;P<0.01].(5)The invasion number of transfecfion group and blank control group were 121±12 and 452±17 respectively.with a significant eclampsia by regulating proliferation and invasion of trophoblast.

Expression of human leucocyte antigen G on haman placenta and its gene polymorphism in relation to intrahepatic cholestasis of pregnancy / 中华妇产科杂志

0.05).Conclusions The reduced expression of HLA-G on placenta in ICP patients may alter the maternal-fetal immune response and thus be involved in the pathogenesis of this disorder. Dexamethasone can upregulate the expression of HLA-G on placenta.The 14 bp deletion polymorphism in exon 8 of HLA-G gene might not have a significant influence on the development of ICP.