2.Identification of HIV-1 specific T lymphocyte responses in highly exposed persistently seronegative Chinese.
Hong-wei LIU ; Kun-xue HONG ; Jun MA ; Lin YUAN ; Sha LIU ; Jian-ping CHEN ; Yuan-zhi ZHANG ; Yu-hua RUAN ; Jian-qing XU ; Yi-ming SHAO
Chinese Medical Journal 2006;119(19):1616-1621
BACKGROUNDStudies of highly exposed persistently seronegative (HEPS) individuals may provide valuable information on mechanisms of protection and on vaccine design. Cellular immune responses play a critical role in containing human immunodeficiency virus. However, the cellular immune responses in HEPS individuals have not been thoroughly assessed at the entire viral genome level.
METHODSTen HEPS Chinese with a history of frequent penetrative vaginal intercourse (mean frequency, at least once a week), with some unprotected sexual contact occurring in the weeks or days immediately before enrollment, 25 HIV-1 seropositive individuals, 10 HIV-1-seronegative healthy individuals with low-risk sexual behavior and no history suggestive of exposure to HIV-1 infection were enrolled. HIV-1-specific T cell responses were comprehensively analyzed by an interferon-gamma Elispot assay against 770 overlapping peptides spanning all HIV-1 proteins.
RESULTSHIV-1-specific T-cell responses of interferon-gamma secretion were identified in 3 (30%) out of 10 HEPS individuals; the specific cytotoxic T lymphocytes were targeted at Pol (2/10), Env (2/10), and Tat (1/10). HIV-1-specific T-cell responses of interferon-gamma secretion were identified in 20 (80%) out of 25 seropositive intravenous drug users (IDUs), revealing that all HIV-1 proteins and protein subunits could serve as targets for HIV-1-specific CD8(+) T cell responses with 85% recognizing Gag, 80% recognizing Nef, 75% recognizing Pol, 60% recognizing Env, 55% recognizing Vpu, 45% recognizing Vpr, 20% recognizing Vif, 20% recognizing Tat and 15% recognizing Rev in these seropositive individuals. None of the seronegative healthy individuals gave the positive T-cell responses.
CONCLUSIONSAbout 30% of HEPS Chinese mounted HIV-1 specific T cell immune responses. Cell-mediated immunity against HIV-1 may be developed through non-productive infections.
Adult ; Female ; HIV Seronegativity ; immunology ; HIV-1 ; immunology ; Humans ; Interferon-gamma ; biosynthesis ; Male ; Receptors, CCR5 ; genetics ; T-Lymphocytes, Cytotoxic ; immunology
3.Expression of IFN-gamma and its receptor alpha in the peripheral blood of patients with chronic hepatitis C.
Ping ZHANG ; Zhi CHEN ; Feng CHEN ; Min-wei LI ; Jun FAN ; Hui-ming ZHOU ; Ji-hong LIU ; Zhu HUANG
Chinese Medical Journal 2004;117(1):79-82
BACKGROUNDIt has been known that intra-cellular immunity is important for defense against viral infections and this function lies with interferon gamma (INF-gamma). Here we evaluated the role of IFN-gamma system in the pathogenesis of chronic hepatitis C (CHC).
METHODSThe levels of interferon gamma receptor alpha (IFNGR alpha) on the peripheral lymphocyte membrane were assayed with flow cytometry. The plasma concentrations of the cytokines IFN-gamma and IL-10 in CHC patients and normal controls were assayed by enzyme-linked-immunosorbent assay (ELISA). The samples were collected randomly from Xinjiang Autonomous Region, Zhejiang and the northern regions of Jiangsu Province in China.
RESULTSThe levels of IFNGR alpha in CHC patients were significantly lower than that of normal controls (NC), especially among patients during the stable stage (P < 0.001), whereas there were no significant differences between CHC in active and stable stages. Among the patients of the three regions, there were no significant differences between patients from Xinjiang and Zhejiang provinces, but both had statistically significant difference compared with the patients from Jiangsu Province (P < 0.001). Plasma IFN-gamma and IL-10 concentrations in CHC patients decreased significantly, IFN-gamma in particular, but there were no significant differences in these levels between various stages of the disease. The IFN-gamma/IL-10 (Th1/Th2) ratio in patients was reversed.
CONCLUSIONThere may be defects in the IFN-gamma system in chronic HCV infected subjects and a low immune response, which may play an important role in the persistence of HCV infection.
Adolescent ; Adult ; Aged ; Child ; Female ; Hepatitis C, Chronic ; blood ; immunology ; Humans ; Interferon-gamma ; blood ; Male ; Middle Aged ; Receptors, Interferon ; blood
4.Modulation and implications in pathogenesis of interferon gamma receptor 1 in patients with chronic hepatitis B.
Gong-ying CHEN ; Zhi CHEN ; Min-wei LI ; Hui-min LIU ; Zhong-rong HU
Chinese Journal of Hepatology 2003;11(1):14-16
OBJECTIVETo study the possible differences in the interferon gamma receptor (IFN-gamma R1) response among a variety of clinical types in patients with chronic hepatitis B (CHB) and implications in pathogenesis.
METHODSThe serum level of IFN-gamma and the expression level of IFN-gamma R1 in peripheral leucocytes, from 53 CHB patients, were examined by ELISA and flow cytometry respectively, which were compared with the baseline levels of 15 healthy controls and were performed correlation analysis with alanine aminotransferase (ALT), total bilirubin (TBil) in serum and morphological change in hepatic tissues.
RESULTSThe results showed that the level of IFN-gamma R1 (28.89% 11.77%) expressed on the membranes of lymphocytes in CHB patients, which correlated with liver inflammation (r=0.621, P<0.01) and serum TBil level (r=0.575, P<0.01), was much higher than that (9.23% 1.30%) of the healthy controls (Z=3.988, P<0.05), and no obvious difference on the membranes of leucocytes. The serum levels of IFN-gamma in patients with cirrhosis and severe hepatitis were higher than those of healthy controls. And the two was no difference from each other, but the standard deviation in each group was relatively large.
CONCLUSIONThese findings suggest that the IFN-gamma signal transduction pathway is modulated through up-regulating the expression of IFN-gamma R1 on the membranes of lymphocytes, which takes part in the immuno-pathogenesis in CHB patients.
Adult ; Aged ; Female ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; blood ; Lymphocytes ; chemistry ; Male ; Middle Aged ; Receptors, Interferon ; blood
5.Antitumor effect of a triple negative breast cancer-dendritic cell fusion vaccine.
Peng ZHANG ; Ruilei LIU ; Hua JIANG ; Yu LIU ; Qiaochu ZHANG ; Yong HUANG
Journal of Southern Medical University 2012;32(6):778-783
OBJECTIVETo test the antitumor effect of a human triple-negative breast cancer cell-dendritic cell (DC) fusion vaccine.
METHODSDCs were isolated from fresh peripheral blood of healthy donors. The fusion vaccine was prepared by fusing the DCs and MDA-MB-231 cells via electrofusion. The morphology of the vaccine was identified under inverted fluorescence microscope and the phenotypes were analyzed with flow cytometry. The production of interleukin-12 (IL-12) and interferon-γ (IFN-γ) by the fusion cells was assessed using ELISA. A CCK-8 kit was used to examine the effect of the vaccine in stimulating the proliferation and cytotoxicity of autologous T lymphocytes.
RESULTSThe DCs isolated from peripheral blood mononuclear cells highly expressed CD83, CD86, CD11c and HLA-DR on the cell surface. The fusion cells were irregular in shape and coexpressed the phenotypes of DCs and MDA-MB-231 cells. The fusion cells possessed a strong ability to stimulate the proliferation of T lymphocytes in vitro. Compared with the control group, the fusion vaccine showed a stronger antitumor effect against the breast cancer cells.
CONCLUSIONThe triple-negative breast cancer-DC fusion vaccine prepared by electrofusion can stimulate the proliferation of T lymphocytes and induces strong cytotoxicity of the T cells against breast cancer cells.
Breast Neoplasms ; immunology ; Cancer Vaccines ; immunology ; Cell Fusion ; Cell Line, Tumor ; Dendritic Cells ; immunology ; Female ; Humans ; Interferon-gamma ; immunology ; Interleukin-12 ; immunology ; Lymphocyte Activation ; T-Lymphocytes, Cytotoxic ; immunology
6.Effects of mild moxibustion on imbalance of Th1/Th2 in rabbits with atherosclerosis and its regulating rules.
Shan WU ; Ling-Ling WANG ; Hai-Hong CAI ; Wen JIANG
Chinese Acupuncture & Moxibustion 2014;34(2):163-167
OBJECTIVETo observe effects of mild moxibustion and lovastatin on immunologic function in rabbits with chronic hyperlipidaemia and atherosclerosis (AS) to initially explain regulating rules of mild moxibustion on immunologic function.
METHODSAmong thirty-two Japanese male big-ear rabbits, 8 rabbits were randomly selec ted as a blank group, the rest 24 rabbits were fed with method of endothelial injury and high-fat diet to establish AS model. The blank group was raised with normal diet and free water. After ten weeks of model establishment, the rest 24 rabbits were randomly divided into a model group, a moxibustion group and a medicine group, eight rabbits in each one. Moxibustion was applied at "Shenque" (CV 8) and "Zusanli" (ST 36) for 10 min per acupoint per day in the moxibustion group, while intragastric administration of 3.6 mg/kg lovastatin capsule was applied in the medicine group. After treatment, serum was acquired. Spectrophotometry method was adapted to measure cholesterol (TC) and high-density lipoprotein (HDL-C) and evaluated atherosclerosis index (AI), while enzyme-linked immunosorbent assay method was used to measure interferon-gamma (IFN-gamma) and interleukin-4 (IL-4).
RESULTS(1) The serum TC and HDL-C in the model group were significantly higher than those in the blank group, moxibustion group and medicine group (all P < 0.01). The mean value of AI was 1.683 +/- 0.486 in the moxibustion group, which was obviously lower than 20.301 +/- 4.022 in the model group (P < 0.01). (2) The ratio of Th1/Th2 was 0.569 +/- 0.143 in the moxibustion group and 0.445 +/- 0.079 in the medicine group, which were significantly lower than 0.917 +/- 0.255 in the model group (both P < 0.01), but there was no statistically significant difference between the moxibustion group and the medicine group (P > 0.05).
CONCLUSIONThe moxibustion for AS could reduce atherosclerosis index, influence drift and bias of helper T cell and regulate balance between humoral immunity and cellular immunity. As a result, status of relative balance of immunity is acquired, which could slow down the development of atherosclerosis and process of thrombus burst.
Animals ; Atherosclerosis ; immunology ; therapy ; Humans ; Hyperlipidemias ; immunology ; therapy ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Male ; Moxibustion ; Rabbits ; Th1 Cells ; immunology ; Th2 Cells ; immunology
7.Study of levels of IL-18 and IFN-Gamma in serum of patients with condyloma acuminatum.
Journal of Zhejiang University. Medical sciences 2003;32(4):327-329
OBJECTIVETo study the role of interleukin(IL)18 and interferon gamma (IFN-Gamma) in the pathogenesis of condyloma acuminatum(CA).
METHODSDouble antibody sandwich ELISA was used to measure the levels of IL-18 and IFN-Gamma in serum of 30 cases of CA.
RESULTThe serum levels of IL-18 and IFN-Gamma of the CA patients were significantly higher than those of normal controls IL-18 132.00 +/-52.92 microg/L compared with 50.79 +/- 23.48 microg/L, P<0.01 IFN-Gamma:80.83 +/-20.46 microg/L compared with 20.71 +/-11.18 microg/L P <0.01) and IL-18 levels were positively correlated with IFN-Gamma concentrations (r=0.754, t=6.081, P <0.01).
CONCLUSIONThese data suggest that during infection of human papilloma viruses, anti-infective immune responses are activated.
Adult ; Condylomata Acuminata ; immunology ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-18 ; blood ; Male ; Middle Aged
8.Implications of Th1 and Th17 cells in pathogenesis of oral lichen planus.
Sanxiang XIE ; Lei DING ; Zhigang XIONG ; Shengrong ZHU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(3):451-457
Oral lichen planus (OLP) is considered a T cell-mediated autoimmune disease with unknown aetiology. T helper cells appear to play an important role in the pathogenesis of OLP. We investigated the possible role of T helper cells, Th1 and Th17, in the lesions and circulation of patients with OLP. Forty patients with OLP and 15 healthy volunteers were recruited. Double immunofluorescence staining was used to detect Th1 and Th17 cells in the OLP lesions, and intracellular cytokine staining and flow cytometry to evaluate the proportion of Th1 and Th17 cells in peripheral blood. The levels of serum interferon (IFN)-γ and interleukin (IL)-17 were assessed by using an enzyme-linked immunosorbent assay. It was found that Th17 cells, as well as Th1 cells, were present in OLP lesions. The proportion of peripheral Th1 and Th17 cells was significantly increased in patients with OLP. The proportion of Th17 cells in atrophic-erosive OLP was elevated as compared with that in reticular OLP. Serum IL-17 levels in OLP patients were significantly higher than in controls, and those in the atrophic-erosive OLP group were increased as compared with the reticular OLP group. However, the levels of serum IFN-γ were slightly decreased in OLP patients. Our data suggested that Th1 and Th17 cells in the local lesions and peripheral blood may be associated with the pathogenesis of OLP, and that IL-17 may be an important proinflammatory cytokine in OLP. These findings enhance our understanding of OLP pathogenesis.
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9.Specific T cell immune response in chronic hepatitis B patients treated with different doses of recombinant hepatitis B vaccine.
Qi-huan XU ; Xiao-hong ZHANG ; Dong-ying XIE ; Jian-guo LI ; Yu-tian CHONG ; Lin YANG ; Wei-lun LU ; Zhi-liang GAO
Chinese Journal of Experimental and Clinical Virology 2007;21(4):334-336
OBJECTIVETo study the specific cellular immunoresponse of peripheral blood lymphocytes in the chronic hepatitis B patients treated with different doses of recombinant hepatitis B vaccine.
METHODSSeventy-two chronic hepatitis B patients who did not use any anti-HBV drugs within 6 months were randomized into 3 groups (90 micrograms, 60 micrograms, and placebo) in a ratio of 1:1:1. The patients in different groups were treated with different doses of recombinant hepatitis B vaccine in combination with IFN alpha 1b 50 micrograms with 3 times a week for 24 weeks. All patients were followed up for 24 weeks (W24). HBV DNA, HBeAg and liver functions were detected at different time points, and the number of cells that secrete IFN-gamma were detected by ELISPOT.
RESULTSThere were no significant difference in ELISPOT positive ratio among the 3 groups on baseline detection. At W24, 12 cases, 12 cases, and 7 cases showed ELISPOT positive in the group of 90 micrograms, 60 micrograms, and placebo. The proportion of patients who were ELISPOT positive was higher in the groups treated with recombinant hepatitis B vaccine (including the dose of 90 micrograms and 60 micrograms) than that in the placebo group (P=0.0446). HBV DNA turned negative in 6/24 of the patients treated with recombinant hepatitis B vaccine (at both the doses of 90 micrograms and 60 micrograms), and HBeAg/Anti-HBe seroconversion or HBeAg became negative in 7/24 of them. In the placebo group, none of the patients showed undetectable HBV DNA, HBeAg/Anti-HBe seroconversion or HBeAg disappearance. At the 24W of follow up, in the patients who were ELISPOT positive, HBV DNA became undetectable in 4 of the patients treated with recombinant hepatitis B vaccine (at doses of 90 micrograms and 60 micrograms), and HBeAg/Anti-HBe seroconversion or HBeAg disappearance were found in 9 of the cases. In the placebo group, none of the cases showed undetectable HBV DNA, and only 1 case had HBeAg/Anti-HBe seroconversion.
CONCLUSIONThe recombinant hepatitis B vaccine may increase the function of specific T lymphocytes in patients with chronic hepatitis B. There were no significant differences between the patients treated with the dose of 90 micrograms and 60 micrograms hepatitis B vaccine.
Adult ; DNA, Viral ; blood ; Female ; Hepatitis B Vaccines ; immunology ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; biosynthesis ; Male ; Recombinant Proteins ; immunology ; T-Lymphocytes ; immunology ; Vaccines, Synthetic ; immunology
10.Effect of hepatitis B virus C protein on function of natural killer cell in NK-92 cells.
Ying-Xia LIU ; Xing-Hui WEI ; Bing LI ; Zhi-Xiong LUO ; Jing-Jing XIE ; Yan TAN ; Su-Jun LONG ; Bo-Ping ZHOU
Chinese Journal of Experimental and Clinical Virology 2008;22(4):275-277
OBJECTIVETo investigate the influence of hepatitis B virus C protein on the function of natural killer cell.
METHODSRecombinant eukaryotic expression plasmid pHBI-CMV-HBC was constructed and confirmed by double restrictive enzyme digestion and DNA sequencing analysis. Then the recombinant plasmid was transfected into NK-92 cells with lipofectamine encapsuled. The transfected NK-92 cells containing expressive HBV C protein was confirmed by Western Blot analysis. ELISA was employed to determine the IFN-gamma level secreted by NK-92 cells. And finally the cytotoxicities of NK cells were analysed by MTT colorimetry, with the hepatoblastoma cell line (HepG2) as target cell.
RESULTSWestern blotting confirmed the expression of HBV C protein in the NK-92 cells transfected with pHBI-CMV-HBC. NK cytotoxicities and IFN-gamma secretion level of NK-92 cells transfected with recombinant plasmid significantly increased compared to control NK-92 cells transfected with blank plasmid (P < 0.01) and untransfected NK-92 cells(P < 0.01).
CONCLUSIONTransient expression of HBC can increase IFN-gamma secretion and cytotoxicities of NK-92 cells.
Cell Line ; Cytotoxicity, Immunologic ; Hepatitis B ; immunology ; virology ; Hepatitis B Core Antigens ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Humans ; Interferon-gamma ; immunology ; Killer Cells, Natural ; immunology