【Abstract】ObjectiveTo inquire into the regulative effect of interferon (IFN) on human leucocyte antigen-DR(HLADR) expression in human colonic cancer cells. MethodsThe expression of HLA-DR in 4 colonic cancer cell lines HCT-8，lovo，SW-480 and Ls-174-T was detected with immunohistochemical SP and ELISA method before and after being stimulated by different doses of α-IFN or γ-IFN. ResultsAll cell lines except lovo cell were HLA-DR positive before stimulation with α-IFN or γ-IFN. HLA-DR expression was enhanced on all of the 4 cell lines after stimulation, and it was correlated with γ-IFN and the dose of γ-IFN. The effect of γ-IFN was more obvious than that of α-IFN. ConclusionIFN can enhance HLA-DR expression in human colonci cancer cells and clinical therapy of IFN for colon cancer has a certain applying prospect.
Abstract: Human leukocyte antigen (HLA) is the key antigen mediating rejection and panel reactive antibody (PRA) represent anti-HLA antibodiesin circulation. HLA typing and PRA testing are carried out generally before organ transplantation. With research on the relationship among HLA, PRA and heart transplantation developing, the value of HLA typing and PRA testing in heart transplantation has received more attention and their clinical using strategy has been improved. This article will review the strategy of HLA typing, the clinical value of HLA typing, time-selection in HLA typing, reason and mechanism of rising PRA, clinical sense of PRA testing and treatment of sensitized patients.
Objective To establish a eukaryotic cell line that can express soluble human leucocyte antigen G1(sHLA-G1) stably. Methods The recombinant plasmid pcDNA3-sHLA-G1 is transfected by a novel nonviral, electroporation-based gene transfer method termed nucleofection into the host cell lymphoblastoid cell line (LCL)721.221 which does not express any HLA-classical I molecules. After selection by G418, the cell line stably expressingsHLA-G1 is identified by RTPCR and Dot-ELISA with HLA-G1 specific monoclonal antibody MEM-G/9. Results The efficiency of transfection for LCL721.221 is about 14% by nucleofection. The specific band forsHLA-G1 was found by RT-PCR assay from the transfections and the protein ofsHLA-G1 in the supernatant of the transfections was detected by Dot-ELISA assay. Both confirmed that the eukaryotic cell line expressingsHLA-G1 has been established successfully at genic and proteinic levels. Conclusion In this study, the eukaryotic cell line expressingsHLA-G1 have been established successfully by nucleofection.
Objective To study whether the porcine endothelial cells (PECs) lines transfected by HLA-G1 can alter the lysis mediated by human peripheral blood mononuclear cell (PBMC) and natural killer cell 92(NK-92). Methods By use of liposomes pack, the pcDNA3.0 eukaryotic expression vector carrying HLA-G1 was transfected into PECs. Using indirect immunofluorescence and RT-PCR assays, the HLA-G1 expression in PECs was detected. The alteration of the lysis mediated by PBMC and NK-92 was detected by51Cr-release assays. Results HLA-G1 expression could be detected in PECs after transfection of HLA-G1 at the levels of protein andRNA. It also could be found that the survival rate of transfected PECs was muchhigher than that of non-transfected PECs, when both of them faced the lysismediated by human PBMC and NK-92.After transfecting the expression of HLA-G1 could be found in the transfected PECs and the lysis mediated by PBMC and NK-92 to PECs decreased obviously (Plt;0.05). Conclusion The PECs- transfected by HLAG1 can decrease the NK lysis, so that it may provide us a new thought to inhibit the xeno-cell-rejection.
Objective To investigate the relation of Human Leukocyte Antigen-DRs to Pre-eclampsia/eclampsia （PE/E） by reviewing the observational studies on PE/E. Methods We searched the MEDLINE/PubMed, EMBASE, The Cochrane Library and CBMdisc to July 2005, by combining free text with MeSH words. We assessed the quality of included studies, extracted and analyzed data. Results The odds ratio of fetal-maternal HLA-DR4 antigen frequency in case group versus control group was 2. 60 （95% CI 1.87 to 3.60） with statistical significance. The antigen frequencies of＇other fetal-maternal HLA-DRs in case and control groups were not statistically significant. The antigen frequencies of the couple HLA DRs were not statistically significant between case and control groups. We found that neither the HLA-DR sharing between the couples nor between fetus and mothers in case and control groups were statistically significant. Conclusions The antigen frequencies of HLA-DRs between the couples may have no association with the development of PE/E. The fetal gene types may be related to the development of PE/E. The HLA-DR sharing in mothers and fetus and the couples may have no association with the development of PE/E.
Objective To investigate the expression of human leukocyte antigen-G （HLA-G） in hepatocellular carcinoma （HCC） tissues， and to evaluate the prognosis of patients after liver transplantation. Methods The clinical data of 83 patients with HCC who underwent liver transplantation from January 2004 to May 2008 in the Liver Transplantation Center of the Third Affiliated Hospital of Sun Yat-sen University were analyzed retrospectively. The expression of HLA-G in HCC tissues was detected by using immunohistochemical analysis. The Kaplan-Meier method was used to evaluate the cumulative survival rate and tumor-free survival rate. Log-rank test and Cox regression model were used to analyze the single and muti-factor for tumor-free survival rate, respectively. Results Among the 83 patients，there were tumor recurrence in 35 patients （42.2%）. The 1-year，3-year, and 5-year of cumulative survival rate was 97.2%，89.8%, and 43.1%, respectively. The 1-year，3-year， and 5-year of tumor-free survival rate was 93.6%，68.9%, and 38.7%, respectively. The positive rate （68.7%） of HLA-G expression in HCC tissues was significantly higher than that in paracancerous tissues （15.7%）, P＜0.01. A significant association was found between expression of HLA-G and tumor size， vascular invasion, and pathology differentiation （P＜0.05）. Single factor analysis showed that the expression of HLA-G （P＜0.01）， tumor size （P＜0.05）， vascular invasion （P＜0.01）， and pathology differentiation （P＜0.01） effected on tumor-free survival rates of HCC patients after liver transplantation. The tumor-free survival rate in positive expression of HLA-G group was significantly lower than that in negative expression of HLA-G group （P＜0.01）. Cox regression model analysis showed that the expression of HLA-G （P＜0.05）， vascular invasion （P＜0.01）， and pathology differentiation （P＜0.05） were independent risk factors that affected the tumor-free survival rate of HCC patients after liver transplantation. Conclusions There is expression of HLA-G in HCC tissues. The independent risk factors that affecting the tumor-free survival rate of HCC patients after liver transplantation include the expression of HLA-G， vascular invasion, and pathological differentiation. Taking interferential measures for patients with positive expression of HLA-G and strict selection of indication of liver transplantation for HCC can reduce the recurrence rate of tumor.
Objective To construct the expression vector of HLA-G-shRNA and investigate the effect of HLA-GshRNA from NK cell lysis. Methods Four HLA-G shRNA plasmids were constructed and transiently transfected to Bel-7402 cell lines， the levels of mRNA and protein of HLA-G were detected by Real-Time PCR and Western blot. The cytotoxicity of NK-92MI cells against the transfected cells was analyzed by LDH releasing assay. Results The gel electrophoresis and sequencing showed that the inserted sequence was identical to the one which we designed, and no aberrations such as mutation，deletion or insertion occurred. The expressions of HLA-G confirmed by Real Time-PCR and Western blot were significantly down-regulated. Bel-7402 cell lines transfected HLA-G shRNA showed higher lytic activity （P＜0.01）. After KIR2DL4 receptor blocked，lytic activity of NK-92 MI cell were decreased （P＜0.01）. Conclusions HLA-G shRNA plasmids are successfully constructed and HLA-G down-regulated can increase NK cytolysis against Bel-7402 cell. After HLA-G combines with KIR2DL4 receptor at the surface of NK cells， the inhibition effect is transferred.
【Abstract】ObjectiveTo develop a new singletube polymerase chain reaction amplification (ST Amp) protocol for the efficient sequencing-based typing (SBT) of human leukocyte antigen DRB1（HLA-DRB1）.MethodsA set of 7 group-specific exonic 5′ amplification primers and a single generic 3′ primer were included together in a single PCR mix to facilitate a single PCR amplification per sample for HLA-DRB1 typing.ResultsAll samples were successfully typed, the typing result was accurate and repeatable.ConclusionST Amp technique has resulted in the ability to perform high-resolution, high-specificity and high-throughput HLA-DRB1 typing by DNA sequencing.
Objective To explore risk factors of stroke-associated pneumonia (SAP) for elderly stroke patients in ICU, and analyze the predictive value of human leukocyte antigen-DR (HLA-DR) on monocytes for SAP. Methods During January 2015 to August 2016, 155 elderly patients with stroke were recruited. The level of monocyte HLA-DR expression was measured after admission and the incidence of SAP was recorded. The risk factors for SAP were analyzed by univariate and multivariate analysis. ROC curve was drawn to analyze prognostic value of HLA-DR. Results SAP occurred in 75 cases with occurrence rate of 48.4%, including 42 early-onset cases and 33 later-onset cases. Age (OR=11.532), Glasgow Coma Scale (OR=7.124), dysphagia (OR=8.846), mechanical ventilation (OR=15.184), atrial fibrillation (OR=7.869), smoking history (OR=11.784), diabetes (OR=7.185) were independent risk factors (all P<0.05). The expression rate of monocyte HLA-DR in the SAP patients was significantly lower than those in the patients without SAP (allP<0.05). Through the ROC curve analysis, the expression rate of HLA-DR that below 78.65% was the optimum cut-off value for prediction of SAP with the area under ROC curve of 0.922, the sensitivity of 80.0% and the specificity of 85.0%. The sensitivity to predict early-onset SAP was 90.5% (38/42), and to predict later-onset SAP was 66.7% (22/33). Conclusions Age, severe coma, dysphagia, mechanical ventilation, atrial fibrillation, smoking history and diabetes are risk factors for SAP in elderly stroke patients in ICU. The detection of monocyte HLA-DR has reference value for early prediction of SAP especially for early-onset SAP with higher sensitivity.