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Objective:To examine the expression of human leukocyte antigen G (HLA-G) in the peripheral blood and cancerous tissues of patients with papillary thyroid carcinoma (PTC) .Methods:The expression of soluble HLA-G (sHLA-G) in the peripheral blood of 50 individuals with PTC (PTC group) , 25 patients with benign thyroid tumors (BTT group) from Department of Thyroid and Breast Surgery, Beilun branch of the First Affiliated Hospital of Zhejiang University and 20 healthy controls (healthy control group) from physical examination center was assessed by ELISA. Immunohistochemical examination of HLA-G levels was also performed on tissue specimens from patients in the PTC and BTT groups, and their correlation with clinicopathological features of thyroid cancer was analyzed. SPSS 19.0 was used for statistical analysis. The measurement data of normal distribution were tested by two independent samples t test. Chi square test was used to compare the rates between the two groups. Results:The sHLA-G expression in peripheral blood was 21.33 (±5.54) , 22.73 (±4.99) , and 18.29 (±4.43) ng/mL in the preoperative PTC, BTT, and healthy control groups, respectively. Compared to the healthy group, sHLA-G levels were considerably higher in the PTC and BTT groups, with statistically significant differences (totally P < 0.05) . There was no significant difference in statistically sHLA-G levels between the BTT and PTC groups ( P > 0.05) . The positive HLA-G expression rate in PTC tissues was 78% (39/50) . There was no evidence of HLA-G expression in common tissues adjacent to PTC. HLA-G was not expressed in benign tumors. HLA-G was linked with the PTC tumor diameter, and the rate of positive expression was considerably greater with tumor diameters >1 cm than with those ≤1 cm ( P<0.05) . The rate of HLA-G positive expression was not significantly correlated with sex, age, multiple foci, extra-glandular invasion, metastasis of lymph nodes, or the TNM stage in PTC individuals ( P > 0.05) . Conclusions:HLA-G is significantly expressed at high levels in PTC tissues, is correlated with the tumor diameter, and may probably have a significant role in this disease. Peripheral blood sHLA-G may be associated with thyroid tumorigenesis, and its value in PTC requires further verification.
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Background: Vitiligo is a pigmentary skin disorder characterised by a chronic and progressive loss of melanocytes. Although several theories have been suggested to the pathogenesis of vitiligo, an autoimmune process leading to melanocyte destruction appears most likely. Human leukocyte antigen-G is a non-classic, major histocompatibility complex Class I molecule that plays an important role in the suppression of the immune response. Several recent studies have provided evidences that polymorphisms in the human leukocyte antigen-G gene might be related with autoimmune diseases. Objectives: The aim of this study was to decide whether exonic single nucleotide polymorphisms in human leukocyte antigen-G contribute to the risk of developing non-segmental vitiligo in the Korean population. Methods: To evaluate the associations between exonic single nucleotide polymorphisms (rs1630223 [Ala5Ala] and rs12722477 [Leu134Ile]) of human leukocyte antigen-G and vitiligo, 244 patients with vitiligo and 398 healthy controls were recruited. Genotyping was performed using Fluidigm 192.24 Dynamic Array with EP1 (Fluidigm Corp., CA). The SNP type assay (Fluidigm Corp., CA), which employs allele-specifically designed fluorescences (FAM or VIC) primers and a common reverse primer was applied and the data were analysed using the EP1 single nucleotide polymorphisms genotyping analysis software to obtain genotype calls. Results: Two exonic single nucleotide polymorphisms (rs1630223 and rs12722477) exhibited significant associations with susceptibility and remained a statistically significant association following Bonferroni correction. These two single nucleotide polymorphisms were located within a block of linkage disequilibrium. Haplotypes G-C and A-A comprising rs1630223 and rs12722477 demonstrated a significant association with non-segmental vitiligo. Limitations: The protein expression level of patients with vitiligo and controls was not studied and a replication study of the genetic association in an independent group was not managed. Conclusion: Our results suggest that exonic human leukocyte antigen-G polymorphisms (rs1630223 and rs12722477) are associated with the development of non-segmental vitiligo.
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RESUMEN El trasplante de órganos sólidos se ha considerado el fin último para algunas enfermedades crónicas en estadio terminal, sin embargo, las incompatibilidades del HLA entre el donante y el receptor pueden permitir que la alorespuesta se convierta en nociva para el órgano trasplantado, respuesta que puede ser tanto innata como adaptativa. Se ha identificado el HLA-G como una molécula natural inductora de tolerancia (28) principalmente en el embarazo y se considera una molécula del HLA clase I no clásico, sin embargo, comparte algunas características estructurales con el HLA clásico. Los genes HLA-G se caracterizan por tener un limitado polimorfismo y una distribución celular y tisular restringida al trofoblasto fetal y células del epitelio tímico entre otras. La búsqueda persistente de la tolerancia en los trasplantes de órganos ha permitido un estudio específico del HLA-G, como posibilidad terapéutica para aumentar la sobrevida tanto de los injertos como de los pacientes trasplantados, es por tal motivo que se realiza una revisión en dicha molécula para estimular la investigación y entendimiento de sus funciones.
ABSTRACT Solid organ transplantation has been considered the ultimate goal for some end-stage chronic diseases, however, HLA incompatibilities between the donor and the recipient may allow the alloresponse to become deleterious for the transplanted organ, a response that can be both innate and adaptive. HLA-G has been identified as a natural tolerance-inducing molecule (28) mainly in pregnancy and is considered a non-classical HLA class I molecule; however, it shares some structural characteristics with classic HLA. HLA-G genes are characterized by having a limited polymorphism and a cellular and tissue distribution restricted to the fetal trophoblast and thymic epithelial cells, among others. The persistent search for tolerance in organ transplants has allowed a specific study of HLA-G, as a therapeutic possibility to increase grafts and transplant patient's survival; for this reason we carried out a review ofthis molecule to stimulate research and understanding of its functions.
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Objective:To analyze the expression of human leukocyte antigen G (HLA-G) in human T-cell leukemia virus type 1 (HTLV-1)-positive T cells, and to investigate its role in the occurrence and development of HTLV-1 infection.Methods:The expression of HLA-G in HTLV-1-positive T cell lines (MT2 and MT4) was detected by Western blot and real-time PCR. HLA-G gene in MT2 and MT4 cells was knocked down by siRNA, and the effects of HLA-G on the expression of HTLV-1 Tax and P19 at mRNA and protein levels were detected by Western blot and real-time PCR. Moreover, the changes in cytokine expression in MT2 and MT4 cells were monitored at RNA level after HLA-G gene silencing. The proliferation ability of MT2 and MT4 cells was analyzed by CCK8. Signal transducer and activator of transcription 3 (STAT3) pathway-related proteins were detected by Western blot.Results:Compared with HTLV-1-negative T cells (Jurkat and MOLT4), the expression of HLA-G increased significantly in MT2 and MT4 cells. After knocking down the HLA-G gene with siRNA in MT2 and MT4 cells, the expression of HTLV-1 Tax and P19 at mRNA and protein levels was decreased, and the expression of antiviral cytokines IFN-γ and TNF-α was increased. The proliferation of MT2 and MT4 cells and STAT3 phosphorylation in these cells were decreased.Conclusions:HTLV-1 could induce T cells to overexpress the immune tolerance molecule HLA-G. Silencing HLA-G gene in HTLV-1-positive T cells could promote the production of antiviral cytokines and reduce IL-6 expression and STAT3 phosphorylation, thereby effectively inhibiting the replication of HTLV-1.
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【Objective】 To investigate the effect of HLA-G expressed in platelets on Tax protein of human T cell leukemia type 1 virus (HTLV-1). 【Methods】 Platelets were isolated from anticoagulant whole blood, and HLA-G molecule on platelet membrane was detected by flow cytometry. The content of secretory HLA-G before and after platelet lysis was detected by ELISA, HTLV-1 human lymphoma cells MT2 were cultured with platelet lysate (PL). The effect of HLA-G in platelets on the expression of HTLV-1 protein Tax was evaluated by Western blot (WB). 【Results】 Membrane type mHLA-G was highly expressed on the surface of platelet membrane. The expression of secretory sHLA-G (ng/mL) increased after platelet lysis (15.73±1.01) vs (6.65±0.47), the expression of sHLA-G increased with the increase of platelet concentration in a dose-dependent manner. Compared with fetal bovine serum, PL significantly promoted the high expression of HLA-G protein and HTLV-1 virus tax protein in MT2 cells, and the addition of anti-HLA-G antibody to PL could effectively inhibit the expression of Tax and HLA-G protein. 【Conclusion】 High expression of immune tolerance molecule HLA-G on platelets can induce high expression of HTLV-1 protein Tax in human lymphoma cell MT2, which contributes to viral infection.
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Objective@#To investigate the relationship between the polymorphism of upstream regulatory region (URR) in human leukocyte antigen-G (HLA-G) gene and the pathogenesis of severe preeclampsia.@*Methods@#Thirty-nine gravidas with severe preeclampsia, who were admitted to the Third Affiliated Hospital of Zhengzhou University from October 2008 to March 2009, were enrolled as the case group. Another 43 healthy gravidas at the third trimester were chosen as the control group. All gravidas in both groups were Han nationality. URR polymorphism in HLA-G gene was detected by PCR sequencing. Allele frequencies and genotype frequencies of all single nucleotide polymorphisms (SNPs) were compared between the two groups.@*Results@#(1) Eight SNPs were detected between -1179 and -689 in HLA-G gene URR in Chinese Han pregnant women. (2) In the severe preeclampsia group, the genotype frequency of -964GG was 12.8% (5/39), which was significantly lower than the frequency of 37.2% (16/43) found in the control group (P=0.012). The allele frequency of -964G was also significantly lower in the severe preeclampsia group than in the control group [38.5% (30/78) vs 57.0% (49/86), P=0.018). (3) In the severe preeclampsia group, the genotype frequency of -716TT was 17.9% (7/39), which was significantly lower than the frequency of 41.9% (18/43) found in the control group (P=0.019). In the severe preeclampsia group, the allele frequency of -716T was 39.7% (31/78), which was also significantly lower than the frequency of 60.5% (52/86) found in the control group (P=0.008). (4) No significant differences were found in the allele or genotype frequencies of -1140A/T, -762C/T or -725C/G in HLA-G gene URR between the two groups (P>0.05).@*Conclusion@#Some of the SNPs in HLA-G gene URR are associated with the susceptibility of severe preeclampsia in Chinese Han gravidas. Pregnant women carrying -964G or -716T may have reduced risk of severe preeclampsia.
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Objective To investigate the roles of human leukocyte antigen-G ( HLA-G) in mye-loid-derived suppressor cell (MDSC) proliferation and M1/M2 macrophage differentiation in C57BL/6-NCI-H446-G tumor-bearing mice for better understanding the mechanisms of HLA-G involved in tumor immune evasion. Methods NCI-H446 ( human small cell lung cancer cells) and NCI-H446-G ( NCI-H446 cells ex-pressing HLA-G) cells were labeled with CFSE at a final concentration of 1μmol/L. CFSE fluorescence lev-els were measured by flow cytometry at different time points. Mouse tumor models were established by subcu-taneous injection of C57BL/6 mice with NCI-H446 and NCI-H446-G cells, respectively. PBS was used to set up negative control group. The mice in each group were sacrificed to collect tissue samples on 5 d, 10 d, 15 d and 20 d after injection. The percentages of splenic CD11b+Gr1+MDSCs, F4/80+CD80+M1 and F4/80+CD206+M2 macrophages were analyzed by flow cytometry. Results Steady expression of HLA-G in NCI-H446-G cells was confirmed by Western blot and flow cytometry. HLA-G enhanced the proliferation of NCI-H446 cells. Tumor size increased dramatically in tumor-bearing mice in the first five days and then de-creased over time. The tumor-bearing mice in the NCI-H446-G group had larger tumor than those in the NCI-H446 group in every time point (P<0. 05) and required longer time to fully reject the tumor. Compared with the PBS and NCI-H446 groups, the percentage of splenic MDSCs in tumor-bearing mice was significantly in-creased in the NCI-H446-G group (P<0. 05). Moreover, the ratio of M1/M2 in NCI-H446-G tumor-bearing mice was much lower than that in the other two groups (P<0. 05). Conclusion This study indicated that HLA-G could increase the percentage of MDSCs and decrease the ratio of M1/M2, which might illustrate the role of HLA-G in tumor immune evasion and its potential clinical significance in cancer immunotherapy.
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Objective To investigate the expression of human leucocyte antigen G (HLA-G) in urothelial carcinoma after renal transplantation,and to analyse the relationship between HLA-G expression and the various clinical and pathological parameters.Methods 29 patients with urothelium carcinoma after renal transplantation for the first time from January 2005 to June 2016 were selected as the experimental group,the age range was 32-70 years,with an average of (55.5 ± 8.1) years.29 non-transplanted patients with urothelial carcinoma as the control group 1,the age range was 36-74 years,with an average of (57.9 ± 8.2) years.15 cases of normal urinary tract epithelial were from cystoscopy biopsy as the control group 2.Immunohistochemical method was used to detect the difference of HLA-G expression between the three groups.The clinical and pathological data of patients with urothelial carcinoma after renal transplantation were analyzed.Results The expression rate of HLA-G was 79.3％ (23/32) in patients with urothelial carcinoma after renal transplantation,37.9％ (11/32) in non-transplanted group and 0 (0/15) in normal urinary tract epithelium group.The expression rate of HLA-G in non-transplanted group was significantly higher than that in normal urinary tract epithelium group (P ＜ 0.05).The expression rate of HLA-G in patients with urothelial carcinoma after renal transplantation was significantly higher than that in nontransplanted group and normal urinary tract epithelium group (P ＜ 0.05).Conclusions HLA-G is associated with the occurrence of urothelial carcinoma after renal transplantation.It may provide a new idea for the prevention and treatment of urinary tract epithelium after renal transplantation.
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[ ABSTRACT] AIM: To investigate the differential expression of human leukocyte antigen-G ( HLA-G) isoforms and its receptors in human monocyte line THP-1 after human cytomegalovirus ( HCMV) infection for exploring the role of HLA-G in HCMV escaping the immune response of the organism .METHODS: THP-1 cells were infected with HCMV Towne strain.The expression of HLA-G isoforms at mRNA and protein levels was determined by RT-PCR and Western blot, respectively.The surface expression of HLA-G and its receptors ILT2/ILT4 and the cell viability were analyzed by flow cytometry.The levels of soluble HLA-G (sHLA-G) and IL-10 were measured by ELISA.RESULTS:After infection of the THP-1 cells with HCMV , no obvious apoptosis in the cells was observed , and the viability of the cells was high .A significant up-regulation of HLA-G1,-G3,-G4 and-G5 at mRNA expression level 1 d after infection was found , while the protein expression of HLA-G1 and HLA-G5 isoforms was mainly detected .The expression of HLA-G/ILT2/ILT4 was evi-dently up-regulated 1 d after infection .The level of sHLA-G was significantly increased 1 d after infection as compared with control group (P<0.01).The expression of IL-10 was obviously up-regulated 1 d post-infection as compared with control group.CONCLUSION:The differential expression of HLA-G isoforms and secretion of the receptors ILT 2/ILT4 and IL-10 in the THP-1 cells are induced after HCMV infection .This study provides experimental evidence for evaluating the immune mechanism of HCMV infection .
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Objective To explore the clinical diagnostic valve of serum soluble human leukocyte antigen‐G (sHLA‐G) in cervical cancer and precancerous lesion cervical intraepithelial neoplasia(CIN) .Methods The serum sHLA‐G level was detected by using ELISA and serum TA‐4 and SCC‐Ag levels were detected by using the light‐emitting electrochemical immunoassay method detec‐ting in 230 cases of cervical carcinoma ,120 cases of CIN and 30 healthy volunteers .The differences among various groups and their relationship with the clinicopathological features of cervical cancer were analyzed .Results (1) The comparison of serum sHLA‐G , TA‐4 and SCC‐Ag levels :there were statistically significant differences in serum sHLA‐G level among various groups (P=0 .000);the serum sHLA‐G level in the cervical cancer group was significantly higher than that in the healthy control group ,CIN Ⅰgroup , CIN Ⅱ group and CIN Ⅲ group (P=0 .000 ,P=0 .000 ,P=0 .002 ,P=0 .006);which in the CIN Ⅲ group was significantly higher than that in the CIN Ⅰ group and the healthy control group (P=0 .001 ,P=0 .021) .There were statistically significant differences in serum TA‐4 level among various groups (P=0 .006);the serum TA‐4 level in the cervical cancer group was significantly higher than that in the healthy control group ,CIN Ⅰ group and CIN Ⅱ group (P=0 .003 ,P=0 .008 ,P=0 .018);which in the CIN Ⅲgroup was significantly higher than that in the healthy control group and the CIN Ⅰ group (P=0 .023 ,P=0 .031) .The differences of serum SCC‐Ag level among various groups had statistically significant differences (P=0 .000);which in the cervical cancer group was significantly higher than that in the healthy control group ,CIN Ⅰ group and CIN Ⅱ group (P=0 .000 ,P=0 .001 ,P=0 .007) , and which in the CIN Ⅲ group was significantly higher than that in the healthy control group and the CIN Ⅰ group (P=0 .013 , P=0 .021) .(2) The relationship between serum sHLA‐G and pathological features of cervical cancer :the serum sHLA‐G level had no significant correlation with the age ,tumor size and pathological type (P>0 .05) ,while serum sHLA‐G was closely related with the FIGO stages and lymph node metastasis (P=0 .008 ,P=0 .031) .The serum sHLA‐G level in the FIGO stage Ⅲ and Ⅳ was significantly higher than that in the FIGO stageⅠ and Ⅱ (U=7 .125 ,P=0 .008) ,and which in the patients with lymph node me‐tastasis was significantly higher than that without lymph node metastasis (U=4 .651 ,P=0 .031) .Conclusion The detection of ser‐um sHLA‐G level can contribute to the early diagnosis and disease condition evaluation of cervical cancer and CIN Ⅲ ,thus which is likely to become a new indicator of early diagnosis of cervical cancer .But its specificity with the occurrence of cervical cancer and precancerous lesion remains to be further investigated by related research .
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Objective To investigate the changes of human leukocyte antigen-G (HLA-G) protein expression and Th1/Th2 type cytokines in intrahepatic cholestasis of pregnancy (ICP) and their relativity to the etiology of ICP. Methods Peripheral blood and placental tissues were obtained from 26 ICP patients (the ICP group) and 22 normal pregnant women (the NP group) in the operation room for Cesarean birth. Immunohistochemistry was used to detect the expression of HLA-G protein in the placental tissues. Meanwhile we tested the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-4 (IL-4) by enzyme-linked immunosorbent assay (ELISA) in the peripheral blood and checked the levels of TBA in the serum.Results TBA level in the ICP group was (27.05±6.08) μmol/L, significant higher than that in the NP group (4.35±2.68)μmol/L (P＜0.01). The positive expression of HLA-G protein in extravillous trophoblast in the ICP group was significantly lower than that in the NP group (P＜0.01). The mean optical density (MOD) of positive expression of HLA-G protein in the placenta tissues in the ICP group (52.91±7.19) was significantly lower than that in the NP group (69.26±7.72) (P＜0.01). The concentration of TNF-α was significantly higher in the ICP group (101.31±19.30) pg/mL than that in the NP group (54.51±23.72) pg/mL (P＜0.01). The concentration of IL-4 was lower in the ICP group (22.16±6.55) pg/mL than that in the NP group (31.69±8.25) pg/mL (P＜0.01). The ratio of TNF-α/IL-4 was higher in the ICP group (4.52±1.91) than that in the NP group (1.72±0.61) (P＜0.01). There was a negative correlation between the MOD of HLA-G protein and TNF-α (r=-0.98, P＜0.01) in the ICP group. No correlation with IL-4 and TNF-α/IL-4 was seen (P＞0.05). There was a positive correlation between TBA and TNF-α (r=0.99, P＜0.01), and a negative correlation between TBA and the MOD of HLA-G protein (r=-1.00, P＜0.01) in the ICP group. No correlation with IL-4 and TNF-α/IL-4 was seen (P＞0.05). Conclusion There is an imbalance of Th1/Th2 cytokines to the Th1 type in the peripheral blood of ICP patients. The expression of HLA-G protein in the placenta of ICP patients decreases, leading to an increase of Th1 type cytokines that may be one of the reasons for liver destroy in ICP.
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Human leukocyte antigen G (HLA-G) is one of the molecules implicated in immunotolerance. To investigate the role of HLA-G in primary cutaneous malignant melanoma (CMM), a series of 47 skin melanocytic lesions were immunohistochemicaily evaluated. The correlation between HLA-G expression and CMM clinicohistopahtologicai data and Bcl-2 expression was also analyzed.HLA-G expression was detected in a variety of cell types. No significant difference in HLA-G expression was observed between malignant and non-malignant melanocytic lesions. HLA-G expres- sion was significantly correlated with the inflammatory infiltration and Bel-2 expression, whereas no significant correlation with ulceration, tumor thickness, clinical stage, histopathologicai subtypes were observed. HLA-G expression may be the result of host immune reaction in tumor microenvi- ronment rather than a malignant feature of CMM.
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OBJECTIVES: To determine the expression of HLA-G and IL-10 and their correlation in tissue of cervical cancer, and to investigate the relationship between their expression and clinicopathologic factors in patients with cervical cancer. METHODS: Tissue samples were obtained from 40 patients diagnosed with cervical cancer and 15 patients with normal cervix for control from Oct. 2004 to Oct. 2005. Quantitative real-time RT-PCR for HLA-G mRNA and semi-quantitative RT-PCR for IL-10 mRNA were used. And proteins of HLA-G and IL-10 were detected by Western blot analysis. RESULTS: Both HLA-G and IL-10 mRNA expression in cervical cancer tissue were higher than normal control, which was statistically significant (P<0.001, P<0.001). The proteins levels of HLA-G and IL-10 in cancer group were also significantly higher than control (P<0.001, P=0.041). The mRNA expression of HLA-G tended to be correlated with IL-10 expression (P=0.061), although it was not statistically significant. Among clinicopathologic factors of cervical cancer, there was inverse relationship between FIGO stage and mRNA value of HLA-G (P=0.045). CONCLUSIONS: The mRNA and protein expression of HLA-G and IL-10 in cervical cancer were much higher than in controls. These results showed that HLA-G and IL-10 might have an important role of tumorigenesis in patients with cervical cancer. The levels of HLA-G and IL-10 seem to be correlated although it was not statistically significant. High HLA-G mRNA expression could be related in early tumorigenesis since it was associated with early stage cervical cancer.
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Female , Humans , Blotting, Western , Carcinogenesis , Cervix Uteri , HLA-G Antigens , Interleukin-10 , RNA, Messenger , Uterine Cervical NeoplasmsABSTRACT
0.05).However,the patients with increased levels of sHLA-G had higher incidence of central nervous system involvement(P=0.007) and more severe disease activity(P=0.027) in comparison with patients with normal plasma sHLA-G levels.Finally,the expression of plasma sHLA-G was not influenced by the treatment with glucocorticoids,immunosuppressive agents or antimalarials.Conclusion The increased production of sHLA-G indicates that sHLA-G may play an important role in the pathogenesis of SLE.The expression of sHLA-G may be associated with disease activity and severity of lupus patients,but be independence of HLA-G 14bp ins/del polymorphism and drug treatment.
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0.05).Conclusion:The abnormal expression of HLA-G protein in eutopic and ectopic endometrium tissues may play a key role in the pathogenesis of adenomyosis.