BACKGROUNDBoth athletes with intensive exercise and aged people may have weakened immunity against virus infection. This study aimed to evaluate whether people undergoing aerobic exercises including competitive cyclists with moderate training (CMT) and middle-aged people practicing Tai Chi Chuan (TCC) exercise have higher immunity against hepatitis B virus than age-matched sedentary controls including college students (CSC) and middle-aged people (MSC).

METHODSHuman peripheral blood mononuclear cells from competitive cyclists and sedentary controls were stimulated by phytohemagglutinin (PHA) to prepare conditioned medium (MNC-CM) for the assessment of inhibitory effects on hepatitis B surface antigen (HBsAg) expression in human hepatoma Hep3B cells.

RESULTSThe inhibitory effects on the relative HBsAg expression of CMT's and TCC's MNC-CM were greater than those of the controls. The CMT's MNC-CM prepared from 5 microg/ml PHA decreased HBsAg expression to 61.5%, whereas that of CSC remained at 83.8%. Similarly, this expression by treatment of TCC group' MNC-CM was 68.4% whereas that of MSC group was 84.3%. The levels of cytokines such as interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), IFN-alpha and interleukin-1beta (IL-1beta) in the MNC-CM from the CMT and TCC groups were greater than those in the controls. Antibody neutralization of CMT's MNC-CM and addition of recombinant cytokines into CSC's MNC-CM indicated that IFN-gamma, TNF-alpha and IFN-alpha had synergistic effects against HBsAg expression. Similar blocking effect was noted in TCC versus MSC groups.

CONCLUSIONThese results suggest that the immunomodulatory response to suppress HBsAg expression in CMT and TCC with moderate aerobic exercise is greater than that in age-matched sedentary controls.

Adult ; Exercise ; Hepatitis B Surface Antigens ; analysis ; Humans ; Interferon-gamma ; physiology ; Male ; Oxygen Consumption ; Tai Ji ; Tumor Necrosis Factor-alpha ; physiology

OBJECTIVETo study the changes of Hes-1, the target gene of Notch signaling pathway, and its relationship with airway inflammation and remodeling in a rat model of asthma.

METHODSForty-eight rats were randomly divided into an asthma group and a control group. The rats in the asthma group were sensitized and challenged by ovalbumin (OVA), and normal saline was used in the control group. Two groups were further divided into 3 subgroups according to time points after challenging, i.e. 4 weeks, 8 weeks and 12 weeks (n=8 rats each). Pathological changes of lungs were observed by light microscopy and the thickness of bronchial smooth muscle layer (Wam) was measured. The levels of IL-4 and INF-γ in rat serum and bronchoalveolar lavage fluids (BALF) were measured using ELISA. Expression levels of Hes-1 protein and mRNA were determined by immunohistochemistry and quantitative real-time PCR respectively.

RESULTSTogether with the extension of challenging, the Wam of rats in the asthma group increased, a decrease of INF-&gamma; level and an increase of IL-4 level in serum and BALF were also observed, and the differences were statistically significant compared with those in the corresponding control group (P<0.05). Hes-1 protein and mRNA levels also increased gradually after OVA challenging and were higher than those in the control group (P<0.05). The levels of Hes-1 protein and mRNA were positively correlated with Wam and IL-4 in serum and BALF, but were inversely correlated with INF-&gamma; in serum and BALF (P<0.05).

CONCLUSIONSLevels of Hes-1 protein and mRNA increased, which were closely related with the levels of airway inflammatory factors and remodeling of airway smooth muscle. Hes-1 may play an important role in the pathogenesis of asthma.

Airway Remodeling ; Animals ; Asthma ; etiology ; Basic Helix-Loop-Helix Transcription Factors ; analysis ; genetics ; physiology ; Disease Models, Animal ; Homeodomain Proteins ; analysis ; genetics ; physiology ; Interferon-gamma ; analysis ; Interleukin-4 ; analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Transcription Factor HES-1

To investigate the function of B7 co-stimulator in activation and differentiation of T cell, B7 gene was transfected into Raji and Jurkat cells by liposome, B7 expression in tumor cells was detected with flow cytometry, and expression of IL-2, IL-4 and IFN-gammamRNA was detected by RT-PCR. Kinetics of secretion of three cytokines was also analyzed at 4, 12, 20 and 48 hours after gene transfection. The results showed that B7(+) Raji cells could induce mRNA expression of IL-2, IL-4 and IFN-gamma on T cell surface; B7(+) Jurkat cells could induce secretion of IL-2 and IFN-gamma. However, B7(-) Raji and B7(-) Jurkat cells could not induce secretion of cytokines. Kinetics of the three cytokines secretion were different, IL-2 and IL-4 were only detectable after 4 hours of T cell activation, whereas IFN-c was detectable after 12 hours of stimulation. The peak levels of IL-2, IL-4 and IFN-gamma were found at 20 hours after activation. It was concluded that tumor cell lines transfected with B7 gene could enhance their immunocompetence, activating T cell efficiently and B7-1 play more critical role in T cell activation and differentiation.
B7-1 Antigen ; genetics ; physiology ; Cytokines ; genetics ; Humans ; Interferon-gamma ; genetics ; Interleukin-2 ; genetics ; Interleukin-4 ; genetics ; Jurkat Cells ; RNA, Messenger ; analysis ; Transfection

AIMTo investigate if interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), interferon-gamma (IFN-gamma) or tumor necrosis factor-alpha (TNF-alpha) are able to stimulate the level of lipid peroxidation of sperm membranes, either alone or in the presence of leukocytes.

METHODSSemen samples from normozoospermic donors were prepared by density gradient. The sperms were exposed to the indicated cytokines, at physiological and infection-inflammation concentrations, in the absence or presence of leukocytes. Lipid peroxidation of the sperm membranes was determined by measuring malondialdehyde (MDA) and 4-hydroxialkenals (HAE) formation.

RESULTSTNF-alpha, IL-8 and IFN-gamma increased the level of sperm membrane lipid peroxidation when tested at physiological concentrations. At infection-inflammation concentrations, only IL-8 was able to produce a higher effect. When assayed in the presence of leucocytes, IL-8 and TNF-alpha showed a higher effect at infection-inflammation concentrations than at physiological concentrations. Finally, IL-8 showed a higher effect in the presence of leukocytes than in their absence at both physiological and infection-inflammation concentrations. TNF-alpha also showed a higher effect when assayed in the presence of leukocytes than in their absence, but only at infection-inflammation concentrations. There was no effect of IL-6 or IL-10 in any of the tested conditions.

CONCLUSIONSeveral pro-inflammatory cytokines at physiological concentrations increase the level of lipid peroxidation of sperm membranes, which could be important for the sperm fecundation process. However, infection-inflammation concentrations of some cytokines, such as IL-8 and TNF-alpha, either alone or in the presence of leukocytes, could drive the lipid peroxidation of the spermatozoa plasma membrane to levels that can affect the sperm fertility capacity.

Cytokines ; analysis ; Humans ; Inflammation ; physiopathology ; Interferon-gamma ; metabolism ; Interleukins ; metabolism ; Leukocyte Count ; Lipid Peroxidation ; Male ; Reference Values ; Spermatozoa ; physiology ; Tumor Necrosis Factor-alpha ; metabolism

To investigate the value of apoptosis of the allo-antigen specific T cells induced by Fas/FasL pathway in order to prevent GVHD in allo-transplant, the CD34(+) cells were transfected with FasL or not, used as effector cells, mixed with allo-antigen specific T lymphocytes with presence or absence of IFN-gamma or IL-2. After 5 days, apoptosis of T cells was detected by TdT nick end mediated dUTP labeling (TUNEL) and flow cytometry. The effects of IFN-gamma or IL-2 on apoptosis of CD34(+) cells of graft induced by Fas/FasL pathway observed as controls. The apoptosis incidence of T cells was (12.1 +/- 1.5)% when CD34(+) cells transfected with FasL were used as effector cells, that was much higher than that T cells with CD34(+) cells non-transfected (p < 0.01). In the presence of IFN-gamma or IL-2, apoptosis incidence reached to (20.1 +/- 2.3)% or (17.6 +/- 1.3)% respectively (p < 0.01). When sFasL was added to CD34(+) cells freshly isolated or induced with IFN-gamma or IL-2, the incidence or apoptosis of CD34(+) cells was (7.8 +/- 0.8)%, (18.7 +/- 1.6)% (p < 0.01) or (7.9 +/- 1.0)% (P > 0.05) respectively. The results suggest that it is possible to induce apoptosis of the allo-antigen specific T cells in grafts activated by allo-antigen by exogenous Fas ligand expressed on receptor cells and that may hopefully provide a new method to prevent GVHD
Animals ; Antigens, CD34 ; analysis ; Apoptosis ; Cell Communication ; physiology ; DNA, Complementary ; genetics ; Fas Ligand Protein ; Humans ; Interferon-gamma ; pharmacology ; Membrane Glycoproteins ; genetics ; physiology ; Mice ; T-Lymphocytes ; cytology ; physiology ; Transfection ; fas Receptor ; biosynthesis

OBJECTIVETo investigate the relationship between NF-kappa B activity and IFN-gamma gene expression, as well as the histopathological changes following liver transplants, both with and without cyclosporin A (CsA) treatment.

METHODSSixty male Wistar and Thirty male SD rats were subjected to orthotopic liver transplants. Fourty-five of the Wistar rats were used as recipients, and were divided into 3 groups: group I, syngeneic control (Wistar-to-Wistar); group II, acute rejection (SD-to-Wistar); and group III: acute rejection treated with cyclosporin A by intramuscular injection (SD-to-Wistar + CSA). After the liver transplants, electrophoretic gel mobility shift assay was used to analyze NF-kappa B activity in the splenocytes of recipient rats, and RT-PCR was used to measure IFN-gamma gene expression in grafted liver specimens. In addition, histopathological examinations were performed to assess the severity of acute liver rejection.

RESULTSIn group I, low levels of NF-kappa B activity were only detectable on day 5 and 7 post-transplant, and only weak IFN-gamma mRNA expression was observed at all time points. By contrast, both high NF-kappa B activity and high expression levels of IFN-gamma mRNA were detected at all time points in group II. In group III, NF-kappa B activity and IFN-gamma mRNA expression were significantly inhibited, as compared to group II (P < 0.05). A good correlation was found between NF-kappa B activity and IFN-gamma mRNA expression (r = 0.815). In addition, NF-kappa B activity and IFN-gamma mRNA expression mirrored histopathological changes in all three experimental groups.

CONCLUSIONSChanges in IFN-gamma mRNA expression levels after liver transplantation are at least partially due to changes in levels of NF-kappa B activity. CsA appears to downregulate NF-kappa B activity, thus inhibiting IFN-gamma gene transcription. Blocking the NF-kappa B mediated transcription pathway may be of benefit in preventing liver transplant rejection.

Animals ; Cyclosporine ; pharmacology ; Down-Regulation ; Gene Expression ; Graft Rejection ; prevention & control ; Interferon-gamma ; analysis ; genetics ; Liver Transplantation ; Male ; NF-kappa B ; physiology ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Transcription, Genetic ; physiology

We investigated the role of fasting hormones and pro-inflammatory cytokines in cancer patients. Hormones (ghrelin, adiponectin, and leptin) and cytokines (TNF-alpha, IFN-gamma, and IL-6) were measured by ELISA or RIA in lung cancer and colorectal cancer patients before the administration of cancer therapy, and measurements were repeated every 2 months for 6 months. From June 2006 to August 2008, 42 patients (19 with colorectal cancer and 23 with lung cancer) were enrolled. In total, 21 patients were included in the cachexia group and the others served as a comparison group. No significant difference in the initial adiponectin, ghrelin, TNF-alpha, IFN-gamma, or IL-6 level was observed between groups, although leptin was significantly lower in cachectic patients than in the comparison group (15.3 +/- 19.5 vs 80.9 +/- 99.0 pg/mL, P = 0.007). During the follow-up, the patients who showed a > 5% weight gain had higher ghrelin levels after 6 months. Patients exhibiting elevated IL-6 levels typically showed a weight loss > 5% after 6 months. A blunted adiponectin or ghrelin response to weight loss may contribute to cancer cachexia and IL-6 may be responsible for inducing and maintaining cancer cachexia.
Adiponectin/analysis ; Aged ; Antineoplastic Agents/therapeutic use ; Cachexia/*physiopathology ; Colorectal Neoplasms/drug therapy/*metabolism/mortality ; Cytokines/*analysis ; Female ; Follow-Up Studies ; Ghrelin/analysis ; Humans ; Interferon-gamma/analysis/physiology ; Interleukin-6/analysis ; Leptin/analysis ; Lung Neoplasms/drug therapy/*metabolism/mortality ; Male ; Middle Aged ; Peptide Hormones/*analysis ; Prognosis ; Prospective Studies ; Survival Rate ; Tumor Necrosis Factor-alpha/analysis ; Weight Gain ; Weight Loss

To detect effects of B7 co-stimulation on cytokines, especially on IL-2 mRNA and transcription factors NF-kappa B and AP-1, antiB7-1 McAb, antiB7-2 McAb and C TLA-4 Ig were added into mixture lymphocyte reaction (MLR) system to block B7/C D28 signal transduction, IL-2 mRNA and IL-4 mRNA were determined by using competitive PCR and IFN-gamma mRNA by using semi-quantitative PCR. MHC class II molecules and B7 transfectants were used to stimulate CD28(+) T cell, effects of B7 on NF-kappa B and AP-1 were detected by DNA-protein binding assay. The results showed that IL-2, IL-4 and IFN-gamma mRNA were significantly lower when blockade of B7-2 in MLR than blockade of B7-1. Synergistic effects could be seen with combination of two or three antibodies. One to six hours after MLR, tDR7 alone induced NF-kappa B binding activity; cotransfecting B7 no significantly difference at early time point. After 6 hours, induction of tDR7 was decreased whereas B7 prolonged the induction of NF-kappa B till 72 hours. tDR7 alone also upregulated AP-1 binding activity, on the contrary to NF-kappa B, co-transfecting B7-1 and B7-2 decreased AP-1 binding activity within 24 hours. But during 48 - 72 hours, B7 also prolonged the AP-1 binding activity. It is concluded that after MLR, B 7 increased IL-2 secretion by decreasing the degradation of IL-2 mRNA and upregulating IL-2 transcription factors. B7 also induced several kinds of cytokines secretion. Effects of B7-1 and B7-2 had no significant difference on transcription factors. It is suggested that the different functions between B7-1 and B7-2 maybe related to the difference of cell expression and kinetics. To study the molecular mechanism of B7 mediated T cell immune tolerance can help us to design a better clinic schema to prevent transplantation rejection and GVHD.
3T3 Cells ; Animals ; Antigens, CD ; physiology ; B7-1 Antigen ; physiology ; B7-2 Antigen ; CD28 Antigens ; physiology ; Gene Expression Regulation ; Humans ; Interferon-gamma ; genetics ; Interleukin-2 ; genetics ; Interleukin-4 ; genetics ; Lymphocyte Culture Test, Mixed ; Membrane Glycoproteins ; physiology ; Mice ; NF-kappa B ; metabolism ; RNA, Messenger ; analysis ; Transcription Factor AP-1 ; metabolism

BACKGROUNDT lymphocytes and matrix metalloproteinase (MMP) play an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). However, the details of the mechanisms involved are unclear. The aims of this study were to investigate the changes in interferon-gamma (IFN-gamma), interleukin-4 (IL-4), MMP-9, MMP-12 and tissue inhibitor of metalloproteinase-1 (TIMP-1) levels in a smoke-induced COPD rat model and the therapeutic effects of glucocorticoids and N-acetylcysteine.

METHODSMale Wistar rats were exposed to cigarette smoke for 3.5 months. Budesonide or N-acetylcysteine was given in the last month. Lung function was measured at the end of the study. IL-4 and IFN-gamma levels were then determined in bronchoalveolar lavage fluid and lung tissue samples by enzyme-linked immunosorbent assay. The expression of MMP-9, MMP-12 and TIMP-1 mRNA in lung tissue was determined by RT-PCR.

RESULTSIn comparison with the control group, rats exposed to smoke had a significant increase in IL-4 and MMP-12 levels and a significant decrease in IFN-gamma levels. In addition, the IL-4/IFN-gamma ratio and MMP-12/TIMP-1 ratio were both higher. At the same time, the ratio of forced expiratory volume in 0.3 second to forced vital capacity (FEV(0.3)/FVC) and dynamic compliance (C(dyn)) decreased and expiratory resistance (Re) increased. By measuring pulmonary mean linear intercept and mean alveolar numbers, obvious emphysematous changes were observed in the smoke exposed group. After treatment with budesonide, IL-4 and MMP-12 decreased and IFN-gamma increased. The IL-4/IFN-gamma ratio returned to normal, though the MMP-12/TIMP-1 ratio remained unchanged. FEV(0.3)/FVC was significantly higher and Re was significantly lower than that in untreated smoke exposed rats. No significant differences were found in pulmonary mean linear intercept and mean alveolar numbers. After treatment with N-acetylcysteine, IFN-gamma increased and the IL-4/IFN-gamma ratio decreased. The MMP-12/TIMP-1 ratio remained unchanged. Re and C(dyn) both improved obviously. No significant differences were found in pulmonary mean linear intercept and mean alveolar numbers. Correlation analysis indicated that IL-4 levels in lung tissue correlated negatively with FEV(0.3)/FVC (r = -0.53, P = 0.001), IFN-gamma levels in lung tissue correlated negatively with Re (r = -0.63, P = 0.000) and positively with C(dyn) (r = 0.44, P = 0.009), and that the IL-4/IFN-gamma ratio correlated negatively with FEV(0.3)/FVC (r = -0.44, P = 0.010) and C(dyn) (r = -0.42, P = 0.015) and positively with Re (r = 0.58, P = 0.000). Finally, MMP-12 correlated negatively with FEV(0.3)/FVC (r = -0.36, P = 0.026).

CONCLUSIONSCigarette smoke exposure increases IL-4 levels and decreases IFN-gamma levels. This may be the result of smoke-induced changes in lung function. Budesonide can mitigate the changes in IL-4 and IFN-gamma levels induced by smoke exposure. N-acetylcysteine has no effect on IL-4, but increases IFN-gamma levels and brings the IL-4/IFN-gamma ratio back to normal. Cigarette smoke can also promote MMP-12 gene expression and elevate the MMP-12/TIMP-1 ratio. This effect may play a role in smoke-induced emphysema. Budesonide and N-acetylcysteine do not alter the MMP-12/TIMP-1 ratio in this study when given in the late phase of smoke exposure.

Acetylcysteine ; therapeutic use ; Animals ; Forced Expiratory Volume ; Glucocorticoids ; therapeutic use ; Interferon-gamma ; analysis ; physiology ; Interleukin-4 ; analysis ; physiology ; Lung ; pathology ; physiopathology ; Male ; Matrix Metalloproteinase 12 ; Metalloendopeptidases ; genetics ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; etiology ; physiopathology ; Rats ; Rats, Wistar ; Smoking ; adverse effects ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; Vital Capacity

OBJECTIVETo study the expression of Galectin-9 and Tim-3 in lungs of mice with asthma and the effect of rosiglitazone (PPAR-&gamma; agonist) on their expression.

METHODSFortyfive BALB/c SPF female mice were randomized into control group and asthma groups with and without rosiglitazone intervention. After ovalbumin stimulation and rosiglitazone intervention the pathological changes of the lung tissues were observed. Galectin-9 and Tim-3 mRNA levels in lung tissues were determined using RT-PCR. The levels of IL-4 and IFN-&gamma; in peripheral blood were measured using ELISA.

RESULTSThe expression of Galectin-9 and Tim-3 mRNA of lung tissues in the untreated asthma group increased significantly compared with the control and the rosiglitazone treated groups (P<0.05). A significantly increased blood expression of IL-4 and a significantly decreased blood expression of IFN-&gamma; were found in the untreated asthma group compared with the control and the rosiglitazone-treated groups (P<0.05). The expression of Galectin-9 and Tim-3 mRNA was positively correlated with blood IL-4 level (r=0.792, r=0.794 respectively; P<0.05), but negatively correlated with blood IFN-&gamma; level (r=-0.692, r=-0.757 respectively; P<0.05).

CONCLUSIONSGalectin-9 and Tim-3 mRNA levels in lungs increase in mice with asthma and significantly correlate with the levels of blood Th1/Th2 cytokines. This suggests that Galectin-9 and Tim-3 are closely related to inflammatory process in asthma. Rosiglitazone treatment may decrease the expression of Galectin-9 and Tim-3.

Animals ; Asthma ; drug therapy ; immunology ; pathology ; Female ; Galectins ; genetics ; Hepatitis A Virus Cellular Receptor 2 ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Lung ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; PPAR gamma ; physiology ; RNA, Messenger ; analysis ; Receptors, Virus ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Thiazolidinediones ; therapeutic use