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1.
Chin Med J (Engl) ; 126(14): 2652-5, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23876890

ABSTRACT

BACKGROUND: The human leukocyte antigen-G (HLA-G) has been considered to be an important tolerogeneic molecule playing an essential role in maternal-fetal tolerance, upregulated in the context of transplantation, malignancy, and inflammation, and has been correlated with various clinical outcomes. The aim of this study was to investigate the clinical relevance of the expression of membrane HLA-G (mHLA-G), intracellular HLA-G (iHLA-G), and soluble HLA-G (sHLA-G) in the peripheral blood of live kidney transplant recipients. METHODS: We compared the expression of the three HLA-G isoforms in three groups, healthy donors (n=20), recipients with acute rejection (n=19), and functioning transplants (n=30). Flow cytometry was used to detect the expression of mHLA-G and iHLA-G in the T lymphocytes of peripheral blood from subjects in the three groups. Enzyme-linked immunosorbent assays were used to detect sHLA-G in the plasma from the three groups. RESULTS: There were no significant differences in mHLA-G and intracellular HLA-G among the three groups, but the sHLA-G plasma level was higher in the functioning group than in the acute rejection or healthy group. We found a subset of CD4(+)HLA-G(+) and CD8(+)HLA-G(+) T lymphocytes with low rates of mHLA-G expression in the peripheral blood of kidney transplantation recipients. Intracellular expression of HLA-G was detected in T lymphocytes. However, there was no correlation between acute rejection and the mHLA-G or intracellular HLA-G expression. CONCLUSION: sHLA-G was the major isoform in the peripheral blood of live kidney transplant recipients and high sHLA-G levels were associated with allograft acceptance.


Subject(s)
HLA-G Antigens/blood , Kidney Transplantation , Living Donors , Adult , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Middle Aged , T-Lymphocytes/immunology
2.
Biochem Biophys Res Commun ; 395(4): 540-6, 2010 May 14.
Article in English | MEDLINE | ID: mdl-20394727

ABSTRACT

Extracorporeal photopheresis (ECP) is an effective immunomodulatory therapy and has been demonstrated to be beneficial for graft-vs-host disease and solid-organ allograft rejection. ECP involves reinfusion of a patient's autologous peripheral blood leukocytes treated ex vivo with 8-methoxypsoralen and UVA light radiation (PUVA). Previous studies focused only on ECP treatment of recipient immune cells. Our study is the first to extend the target of ECP treatment to donor immune cells. The results of in vitro co-culture experiments demonstrate uptake of donor PUVA-treated splenic lymphocytes (PUVA-SPs) by recipient immature dendritic cells (DCs). Phagocytosis of donor PUVA-SPs does not stimulate phenotype maturation of recipient DCs. In the same co-culture system, donor PUVA-SPs enhanced production of interleukin-10 and interferon-gamma by recipient DCs and impaired the subsequent capability of recipient DCs to stimulate recipient naïve T cells. Phagocytosis of donor PUVA-SP (PUVA-SP DCs) by recipient DCs shifted T-cell responses in favor of T helper 2 cells. Infusion of PUVA-SP DCs inhibited cardiac allograft rejection in an antigen-specific manner and induced CD4(+)CD25(high)Foxp3(+) regulatory T cells. In conclusion, PUVA-SP DCs simultaneously deliver the donor antigen and the regulatory signal to the transplant recipient, and thus can be used to develop a novel DC vaccine for negative immune regulation and immune tolerance induction.


Subject(s)
Dendritic Cells/immunology , Graft Rejection/therapy , Heart Transplantation/immunology , Immunomodulation , T-Lymphocytes, Regulatory/immunology , Animals , CD4 Antigens/immunology , Dendritic Cells/drug effects , Dendritic Cells/radiation effects , Down-Regulation , Forkhead Transcription Factors/immunology , Interferon-gamma/immunology , Interleukin-10/immunology , Interleukin-2 Receptor alpha Subunit/immunology , Methoxsalen/pharmacology , Phagocytosis , Photopheresis , Rats , Rats, Inbred Lew , Rats, Sprague-Dawley , Spleen/immunology , Th2 Cells/immunology , Ultraviolet Rays
3.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 17(6): 1492-6, 2009 Dec.
Article in Chinese | MEDLINE | ID: mdl-20030933

ABSTRACT

The aim of this study was to investigate the immune regulatory effect of dendritic cells phagocytosing photochemotherapy-treated allogeneic spleen lymphocytes on syngeneic T cells. DA rat spleen lymphocytes were treated with 8-methoxypsoralen plus UVA irradiation (PUVA). LEW rat bone marrow-derived DCs were co-cultured with PUVA-treated DA spleen lymphocytes (PUVA-SP), and the surface markers (MHC-II, CD86 and CD40) of treated DC were detected by flow cytometry. CFSE-labeled PUVA SP were incubated with LEW DCs and the phagocytosis of DCs on PUVA-SP was observed by using fluorescent microscope. The ability of DC phagocytosing allogeneic PUVA-SP (PUVA-SP DC) to stimulate the proliferation of LEW T cells was analyzed by mixed leukocyte reactions (MLR). The production of IL-4, IL-10, IL-2, IFN-gamma in MLR culture supernatant was determined by luminex method. The results indicated that the PUVA treatment effectively induced early apoptosis of DA rat spleen lymphocytes. After co-culture, DC efficiently phagocytosed allogeneic PUVA-SP and still maintained an immature phenotype with low levels of MHC II, CD40 and CD86. PUVA-SP DC induced LEW T cell hyporesponsiveness to DA rat antigen, and led to skewing of T cell cytokine expression toward Th2 (IL-10 and IL-4). It is concluded that the PUVA-SP DC effectively down-regulate T cell response to alloantigen and induce Th2 immune deviation in vitro.


Subject(s)
Dendritic Cells/immunology , Dendritic Cells/physiology , Phagocytosis/immunology , T-Lymphocytes/immunology , Animals , Dendritic Cells/cytology , Flow Cytometry , Isoantigens , Photochemistry , Rats , Rats, Inbred Lew
4.
Zhonghua Gan Zang Bing Za Zhi ; 13(2): 128-31, 2005 Feb.
Article in Chinese | MEDLINE | ID: mdl-15727702

ABSTRACT

OBJECTIVE: To determine which expression mode of prothrombin time (PT) might achieve PT standardization in patients with advanced liver diseases. METHODS: PT was measured with six thromboplastins with different ISI values in 16 severe chronic hepatitis patients, 50 decompensated liver cirrhosis patients and 30 patients on oral anticoagulation therapy. The results were expressed in PT (second), PTA (%), PTR and INR. RESULTS: In chronic hepatitis patients, the means of the six group's PTAs ranged from 24% to 34%, while their upper limits ranged from 47% to 61%. The means of the INRs ranged from 2.55 to 5.13, while their upper limits ranged from 4.65 to 12.77. Through one-way ANOVA of repeated measures, PPTA (0.489) was > PINR (0.120). In patients with liver cirrhosis, the means of the PTA in six groups ranged from 50% to 59%, while their upper limits ranged from 82% to 90%. The means of the INR ranged from 1.40 to 1.80, while their upper limits ranged from 1.97 to 3.69. Through one-way ANOVA of repeated measures, PPTA (0.102) was > PINR (0.01). In patients on oral coagulation therapy, the means of PTA ranged from 26% to 37%, while their upper limits ranged from 39% to 49%. The means of INR ranged from 2.35 to 2.66, while their upper limits ranged from 3.16 to 4.26. Through one-way ANOVA of repeated measures, PPTA (0.01) was less than PINR (0.102). The correlation between the results detected by Neoplastine and by other reagents were analyzed. They correlated well with each other when PTA was used as the expression mode of PT in patients with advanced liver disease. But in patients on oral anticoagulation therapy, when only the INR was used as the expression mode of PT, the correlation was well with each other. CONCLUSION: The use of INR provides inadequate standardization. Only when the PT is expressed in PTA, then it may provide a standardization mode in patients with advanced liver diseases.


Subject(s)
Hepatitis, Chronic/blood , Liver Cirrhosis/blood , Liver Failure/blood , Prothrombin Time/standards , Female , Humans , International Normalized Ratio , Male , Reference Standards
5.
Zhonghua Gan Zang Bing Za Zhi ; 13(1): 31-4, 2005 Jan.
Article in Chinese | MEDLINE | ID: mdl-15670488

ABSTRACT

OBJECTIVE: To investigate the relationship between hemostatic changes in liver cirrhosis patients with different degrees of their liver lesions. METHODS: Forty-three patients (35 men, 8 women; age: 25 to 71 yr) with liver cirrhosis were divided into three subgroups (A, B, and C) on the basis of Child-Pugh classification. Among the patients, 13 were classified as Child-Pugh class A, 15 were class B, 15 were class C. 16 healthy individuals served as controls. A series of hemostatic tests and parameters including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib), factors II, V, VII, VIII, IX, X, vWF assay, antithrombin-III (AT-III), protein C (PC), D-dimer, tissue plasminogen activator antigen (t-PA), plasminogen activator inhibitor activity (PAI) were performed on 43 patients and the 16 healthy controls. RESULTS: PT and APTT were progressively prolonged from A to B and then to C. In comparison to the controls there was a significant difference. Fibrinolytic activity and the activities of factors II, V, VII, IX, X were progressively decreased from A to B and then to C. In comparison to the controls there was a significant difference . AT-III and PC activity were progressively decreased from A to B and then to C. In comparison to the controls there was a significant difference. D-dimer and t-PA-antigen were progressively increased from A to B and then to C. In comparison to the controls there was significant difference. PAI activity did not display significant changes in the four groups. CONCLUSION: We found that there is a close relationship between the severity of cirrhosis and the hemostatic changes. Because the deterioration of the coagulation function and increasing fibrinolytic activity parallel the severity of liver cirrhosis, adequate treatment for cirrhotic bleeding should not only correct the coagulation defects, but also lower the increased fibrinolytic activity.


Subject(s)
Hemostasis , Liver Cirrhosis/blood , Severity of Illness Index , Adult , Aged , Antithrombins/metabolism , Blood Coagulation Factors/metabolism , Female , Fibrinogen/metabolism , Hepatitis B, Chronic/blood , Hepatitis B, Chronic/complications , Humans , Liver Cirrhosis/diagnosis , Liver Cirrhosis/etiology , Male , Middle Aged , Prothrombin Time
6.
Zhonghua Gan Zang Bing Za Zhi ; 12(3): 134-6, 2004 Mar.
Article in Chinese | MEDLINE | ID: mdl-15059293

ABSTRACT

OBJECTIVE: To determine the role of Pre-S1 protein in diagnosing viral replication in patients with chronic hepatitis B. METHODS: 104 consecutive patients with chronic hepatitis B were included in the study, liver biopsy were performed in all patients. Serial serum samples were studied with the quantitative determination of HBV-DNA by a quantitative PCR assay, determination of Pre-S1 protein by ELISA. RESULTS: The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg HBeAg anti-HBc (+) both were 96.5%. The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg anti-HBe anti-HBc (+) were 81.5%, 72.3%, respectively. The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg anti-HBc (+) were 87.5%, 75.0%, respectively. It represented some patients with HBeAg (-) anti-HBe (+/-) still had viral replication. HBV-DNA>10(3) copy/ml as positive criteria for diagnosing viral replication, the positive rate of HBeAg, Pre-S1 were 31.5% (28/89), 80.9% (72/89) in patients with HBV-DNA>10(3) copy/ml, respectively. The concordance rates of HBeAg, Pre-S1 with HBV-DNA were 40.0% (42/104), 82.0% (85/104), respectively. CONCLUSION: It showed that Pre-S1 was more sensitive than HBeAg in diagnosing viral replication in patients with chronic hepatitis B.


Subject(s)
DNA, Viral/blood , Hepatitis B Surface Antigens/blood , Hepatitis B e Antigens/blood , Hepatitis B virus/physiology , Hepatitis B, Chronic/virology , Protein Precursors/blood , Virus Replication , Adult , Female , Humans , Male , Middle Aged
7.
Zhonghua Gan Zang Bing Za Zhi ; 12(4): 210-2, 2004 Apr.
Article in Chinese | MEDLINE | ID: mdl-15099468

ABSTRACT

OBJECTIVE: To determine the reason of thrombocytopenia in patients with liver cirrhosis, we studied the relationship among platelet counts, serum thrombopoietin (TPO) level and spleen index. METHODS: Serum TPO, platelet counts and spleen index were measured in 71 cirrhotic patients. TPO was measured with ELISA method, spleen index were measured on ultrasonography by the same doctor. RESULTS: Platelet counts in patients with cirrhosis were lower than that of healthy group [(109.20+/-53.39) vs (169.63+/-26.60) x 10(12)/L, P<0.05]. Serum thrombopoietin level in patients with cirrhosis was similar to that of healthy group [(436.42+/-258.97) vs (412.63+/-132.80) pg/ml, P>0.05]. However, serum thrombopoietin level decreased as liver disease aggravated, [(526.13+/-317.44) pg/ml in Child-Pugh grade A, (445.22+/-214.90) pg/ml in grade B and (311.45+/-182.66) pg/ml in grade C, grade A vs. Grade C, P<0.05]. However, decline in platelet counts was accompanied with incline in spleen index coordinately. 35 of 71 cirrhotic patients had normal platelet counts whereas 36 of them had thrombocytopenia. Thrombopoietin levels were higher in non-thrombocytopenia group than in thrombocytopenia group [(529.43+/-282.64) vs. (351.27+/-228.25)pg/ml, P<0.01]; but spleen index of two groups showed no difference [(29.65+/-12.00) vs. (36.35+/-12.68) cm2, P>0.05]. Correlation was found between thrombopoietin level and platelet counts (r=0.252, P=0.025); no correlation was found between spleen index and platelet counts (r=-0.238, P=0.062). CONCLUSION: The decline serum TPO levels might play an important role for thrombocytopenia in patients with liver cirrhosis.


Subject(s)
Liver Cirrhosis/blood , Thrombopoietin/blood , Adult , Female , Humans , Liver Cirrhosis/pathology , Male , Middle Aged , Platelet Count , Portal Vein/pathology , Spleen/pathology
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