OBJECTIVETo study the effects of huai qi huang, a traditional Chinese medicine, on cytokines Th1, Th2 and Th17 levels and alveolar macrophage phagocytosis in asthmatic rats sensitized by ovalbumin (OVA).

METHODSForty male Sprague-Dawley rats were randomly divided into five groups: normal control, untreated asthma, budesonide-treated, huai qi huang-treated and budesonide+huai qi huang-treated asthma (n=8 each). Asthma was induced by OVA sensitization and challenge. The levels of IL-4, IFN-γ and IL-17 in plasma and bronchoalveolar lavage fluid (BALF) were measured using ELISA. The phagocytosis of alveolar macrophages which were isolated and purified from BALF was evaluated by the colorimetric assay.

RESULTSThe levels of IL-4 and IL-17 increased, in contrast, the IFN-γ level decreased in plasma and BALF in the untreated asthma group compared with those in the normal control group. The IFN-γ level in the huai qi huang-treated asthma group was higher than that in the untreated asthma group. The IFN-γ level increased and the IL-17 level decreased more significantly in the budesonide+huai qi huang-treated asthma group when compared with the budesonide and huai qi huang alone treatment groups. The phagocytosis of alveolar macrophages in the untreated asthma group was lower than that in the normal control group. Huai qi huang alone or combined with budesonide increased the phagocytosis of alveolar macrophages compared with the normal control, untreated asthma and budesonid-treated asthma groups. The levels of IFN-γ in plasma and BALF were positively correlated with the phagocytosis of alveolar macrophages.

CONCLUSIONSThe levels of IL-4 and IL-17 increase and the IFN-γ level decreases in plasma and BALF, and the phagocytosis of alveolar macrophages decreases in asthmatic rats. Huai qi huang treatment may increase the IFN-γ expression in plasma and BALF and the phagocytosis of alveolar macrophages in asthmatic rats. There is a synergistic effect between huai qi huang and glucocorticoids.

Animals ; Asthma ; drug therapy ; immunology ; Cytokines ; biosynthesis ; Macrophages, Alveolar ; drug effects ; immunology ; Male ; Medicine, Chinese Traditional ; Phagocytosis ; drug effects ; Rats ; Rats, Sprague-Dawley ; T-Lymphocytes, Helper-Inducer ; immunology ; Th1 Cells ; immunology ; Th17 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo prepare an O/W ginseng saponins-based nanoemulsion and investigate its amplified immune response.

METHODThe formulation of ginseng saponins-based nanoemulsion was optimized via the range of nanoemulsion zone in phase diagrams and the solubility of ginseng saponins. Its physicochemical properties were investigated, including morphology, particle size distribution, pH, viscosity and stability. Ginseng saponins-based nanoemulsion as adjuvant was co-administrated with a model antigen ovalbumin (OVA) in mice. Two weeks after the boosting, the serum levels of OVA-specific antibody and its isotypes were determined.

RESULTThe optimized ginseng saponins-based nanoemulsion formulation consisted of ginseng saponins, IPM, Cremophor RH 40, glycerol and water (with the weight ratio of 2 : 4 : 17.8 : 17.8 : 58.4), which was a light yellow fluid. The shape of droplets was spherical under transmission electron microscopy with an average diameter of 72.20 nm and a polydispersity index of 0.052. The viscosity and pH value of it were 4.20 s and 6.02, respectively. And it showed good stability. When co-administered with OVA, no obvious side effects were observed in the mice immunized with ginseng saponin-based nanoemulsion. The serum levels of IgG, IgG1 and IgG2a antibody in the group of ginseng saponin-based nanoemulsion immunized mice was significantly increased compared to the groups of OVA and the saline solution of ginseng saponin. Compared with the adjuvant aluminium hydroxide, the serum levels of IgG and IgG1 antibodys in the groups of ginseng saponins-based nanoemulsion had no significant difference, but the level of IgG2a was obviously higher.

CONCLUSIONginseng saponin-based nanoemulsion could amplify the Th1 and Th2 immune responses, and can be used as the vaccine adjuvant.

Animals ; Drug Carriers ; chemistry ; Emulsions ; chemistry ; Female ; Immune System Phenomena ; drug effects ; Immunoglobulin G ; blood ; immunology ; Mice ; Mice, Inbred ICR ; Panax ; chemistry ; immunology ; Particle Size ; Random Allocation ; Saponins ; chemistry ; immunology ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology

This study was purposed to investigate the effect of prostaglandin E2 (PGE2) on proliferation of peripheral blood T lymphocytes, and to evaluate the regulatory role of PGE2 on immunological balance between Th1/Th2 and Tc1/Tc2 lymphocytes. The peripheral blood mononuclear cells (PBMNC) were stimulated by anti-human CD3 monoclonal antibody (mAb) and anti-human CD28 mAb, and were cultured in the presence of different concentration of PGE2 for 120 hours. The proliferation of peripheral blood T lymphocytes was assayed according to the manufacture protocol of BrdU Kit; the IFN-gamma and IL-4 levels in supernatants cultured for 24, 48, 72 and 120 hours were detected by ELISA; the ratios of CD4+IL-4+ T cells/CD4+ IFN-gamma+ T cells and CD8+IL-4+ T cell/CD8+IFN-gamma+ T cells were determined by flow cytometry. The cells cultured without PGE2 were used as control. The results indicated that (1) with the raising of concentration of PGE2, the inhibitory rate of T cell proliferation in vitro significantly increased (p=0.001). There was significant positive correlation between inhibitory rate of T cells and PGE2 concentration (correlation coefficient=0.889, p=0.000). (2) the difference between the IFN-gamma concentrations in supernatant cultured for 120 and 72 hours in test groups had no statistical significance (p=0.917). The IFN-gamma concentration increased continually with prolonging of culture time in control group (p=0.046). The IFN-gamma concentrations produced at different times in test group were significantly lower compared with those in control group (p<0.05). The IL-4 concentrations produced at different time had no significant change in test groups (p=0.400). The IL-4 concentration in 24 hours in control group was significantly higher than that at 48, 72 and 120 hours in control group (p=0.007, 0.003 and 0.002). After cultured for 24 hours the IL-4 concentration in test group was significantly lower than that in control group (p=0.037), but after cultured for 48, 72 and 120 hours, the IL-4 concentration in test group did not show statistical difference in comparison with control group (p>0.05). (3) the proportions of CD4+IFN-gamma+T cells in test group and in control group had no significant difference (p=0.767). The proportion of CD4+IL-4+T cells in test group was slightly higher than that in control group (p=0.051). The ratio of CD4+IL-4+T cells to CD4+IFN-gamma+ T cells in test group was significantly higher than that in control group (p=0.011). The proportions of CD8+IFN-gamma+ T cells in test group and in control group had no statistical difference (p=0.441). The proportion of CD8+IL-4+T cells in test group was significantly higher than that in control group (p=0.015). The ratio of CD8+IL-4+ T cells to CD8+IFN-gamma+ T cells in test group were obviously higher than that in control group(p=0.038). It is concluded that the PGE2 inhibits the proliferation of T lymphocytes in vitro. PGE2 influences the production of IFN-gamma and IL-4, and significantly influences peak appearance of IFN-gamma produced by T lymphocyte. PGE2 can continuously inhibit the production of IFN-gamma, but its continuous effect on IL-4 is no significant. PGE2 enhances the ratio of CD4+IL-4+T lymphocytes to CD4+IFN-gamma+T lymphocytes and the ratio of CD8+IL-4+T lymphocytes to CD8+IFN-gamma+T lymphocytes, and regulates development of T cells toward Th2/Tc2 cells.
Cell Proliferation ; drug effects ; Dinoprostone ; pharmacology ; Flow Cytometry ; Humans ; Lymphocyte Activation ; drug effects ; Lymphocyte Count ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology

OBJECTIVETo discuss the effect of integrated traditional Chinese and Western medicine on Th1/Th2 cytokines level in children with asthma.

METHODChildren with asthma were randomly divided into control group and treatment group. The control group was given Montelukast Sodium tablets for 4 mg (2-5 years old) or 5 mg (6-14 years old), once every night before sleeping. At the same time, the treatment group was given Pingchuan water decoction additionally for one tie per day, in four to six-divided doses for eight weeks. On the other hand, groups of health children were selected as blank control. Before and after treatment, the level of IL-4, IL-13 and IFN-gamma were detected by elisa from 3 mL of venous blood.

RESULTBefore treatment, IL-4 and IL-13 in two groups were obviously higher than healthy group, but IFN-gamma was lower (P < 0.01, P < 0.05). After eight weeks, IL-4 and IL-13 in treatment group went down, but IFN-gamma increased. And there were significant different compared with the control group (P < 0.05).

CONCLUSIONTreating children asthma by integrated traditional Chinese and western medicine can obviously reduce IL4 and IL-13, increase IFN-gamma, and therefore lighten the airway inflammation, prevent asthma attacks effectively.

Adolescent ; Asthma ; drug therapy ; immunology ; metabolism ; Child ; Cytokines ; metabolism ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Th1 Cells ; drug effects ; metabolism ; Th2 Cells ; drug effects ; metabolism ; Western World

OBJECTIVETo investigate immunomodulatory effect of Astragalus polysaccharides (APS) on IL-12-secreting dendritic cell (DC) subset CD11c(high)CD45RB(low) DC.

METHODSSpleen CD11c(high)CD45RB(low) DC and CD4(+)T lymphocytes in BALB/c mice were purified by magnetic beads sorting, and were treated with 0 (as control), 50, 100, 200 µg/mL APS. Immunofluorescence staining and flow cytometry were used to determine expressions of CD11c(high)CD45RB(low) DC surface molecules, including CD40, CD80, CD86, I-A/E, and Toll-like receptor (TLR) 4. IL-12 level in CD11c(high)CD45RB(low) DC culture supernatant was determined by ELISA. The CD4(+) T lymphocytes were divided into: normal control group, non-stimulation group (CD4(+)T lymphocytes cocultured with APS-unstimulated CD11c(high)CD45RB(low) DC), high-dose APS stimulation group (CD4(+)T lymphocytes cocultured with 200 µg/mL APS-stimulated CD11c(high)CD45RB(low) DC), high-dose APS stimulation+antibody 1 group (CD4(+)T lymphocytes cocultured with 200 µg/mL APS-stimulated CD11c(high)CD45RB(low) DC and IL-12 antibody), high-dose APS stimulation+ antibody 2 group (CD4(+)T lymphocytes cocultured with 200 µg/mL APS-stimulated CD11c(high)CD45RB(low) DC and IL-12 antibody isotype). Proliferation ability of CD4(+) T lymphocytes was determined with MTT method. IL-4 level as well as IFN-γ level in CD4(+)T lymphocyte culture supernatant was determined by flow cytometry. Data were processed with one-way analysis of variance.

RESULTSCompared with those in control, the expressions of CD11c(high)CD45RB(low) DC surface molecules (except for CD86) on CD11c(high)CD45RB(low) DC surface, as well as IL-12-secreting level with dose-dependence were increased in cells stimulated with 50, 100, 200 µg/mL APS. Proliferation ability of CD4(+)T lymphocytes in high-dose APS stimulation group was higher as compared with that in non-stimulation group (F = 13.438, P < 0.05). IFN-γ level in high-dose APS stimulation group \[(2784 ± 137) pg/mL\] was higher than that in non-stimulation group \[(1952 ± 101) pg/mL, F = 12.177, P < 0.05\]. IL-4 level in high-dose APS stimulation group was (172 ± 20) pg/mL, which was lower than that in non-stimulation group \[(193 ± 19) pg/mL, F = 11.963, P < 0.05\]. Proliferation ability of CD4(+) T lymphocytes, IFN-γ level, and IL-4 level in high-dose APS stimulation + antibody 1 group were all ameliorated when compared with those in non-stimulation group; while levels of the 3 indexes in high-dose APS stimulation + antibody 2 group were similar to those in high-dose APS stimulation group.

CONCLUSIONSAPS can activate IL-12-producing CD11c(high)CD45RB(low) DC, and further induce the activation of immune function of T lymphocyte with shifting of Th2 to Th1 in vitro. APS can enhance the immune response via promoting the phenotypic and functional maturation of CD11c(high)CD45RB(low) DC.

Animals ; Astragalus Plant ; chemistry ; Cell Differentiation ; Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; secretion ; Interleukin-12 ; metabolism ; Mice ; Mice, Inbred BALB C ; Polysaccharides ; pharmacology ; Th1 Cells ; immunology

The cytokine pattern on viral antigen recognition is believed to exert a profound influence on the resolution of viral infections and viral clearance. This study was initiated to investigate whether a cytokine imbalance oriented toward Th2 type response plays a role in chronic hepatitis B. Cytokine profiles of peripheral blood mononuclear cells associated with chronic hepatitis B were analysed by RT-PCR. Upon HBsAg stimulation, expression of IFN-gamma, IL-2, IL-4, and IL-10 was detected in 41%, 8%, 41%, and 50% of the patients, respectively. Among these cytokines, the expression of IFN-gamma was associated with high levels of serum AST/ALT. However, we could not prove that Th2 type cytokines had a protective effect on hepatocytes. Upon HBxAg stimulation, there was no recognizable association of cytokine patterns with AST/ALT levels. In conclusion, production of a Th1 cytokine, IFN-gamma, by HBsAg-reactive cells was associated with hepatocyte damage in chronic hepatitis B, while no counteracting effect of Th2 cytokines produced by those cells was observed.
Cytokines/genetics ; Cytokines/biosynthesis* ; Hepatitis B Surface Antigens/pharmacology ; Hepatitis B Surface Antigens/immunology* ; Hepatitis B, Chronic/immunology* ; Human ; Interferon Type II/genetics ; Interferon Type II/biosynthesis ; Leukocytes, Mononuclear/immunology ; Liver/cytology ; Recombinant Fusion Proteins/pharmacology ; Recombinant Fusion Proteins/immunology ; Th1 Cells/immunology* ; Th1 Cells/drug effects ; Th2 Cells/immunology* ; Th2 Cells/drug effects ; Trans-Activators/pharmacology ; Trans-Activators/immunology*

Adolescent ; Antibody Formation ; drug effects ; Asthma ; drug therapy ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Powders ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology

OBJECTIVETo evaluate the Th17/Th1 response in HIV infected patients and the mutual relationship between the response of Th17 and Th1.

METHODS38 chronic HIV infected patients as well as 24 healthy volunteers were performed in this study. The patients were divided into two groups, one group before treatment, the other after therapy. The whole blood intracellular cytokine staining was used, samples detected by BD FACSCanto, after that, the expression of CD4+ IL-17+ T cell and CD4 IFN-gamma+ T cell were analyzed by FACSDiva software and lastly compared the differences among different groups.

RESULTSThe expression of CD4+ IL-17+ T cell in naive-therapy patients were significantly lower than that of the healthy controls (1.14 +/- 0.7)9% vs (3.98 +/- 1.14)%, P = 0.000, but increased remarkably after HARRT(highly antiretroviral treatment) (2.22 +/- 1.00)%, P = 0.001; however there were no significant differences in the expression of CD4+ IFN-gamma+ T cell before and after therapy (34.35 +/- 24.38)% vs (42.10 +/- 15.57%), also with the healthy control (P = 0.383). The frequency of CD4 IL-17+ T cell was positively correlated with CD4+ T counts (R = 0.345, P = 0.034), but no significant correlations was observed between the expression of CD4+ IFN-gamma T cell and CD4+ T counts (R = -0.247, P = 0.136).

CONCLUSIONThe infection of HIV virus down-regulated Th17 immune response and disturbed the balances between Th17 and Th1 evidently in human. Th17 response may play an important role in the pathogenesis of HIV infection.

Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Case-Control Studies ; Female ; HIV ; immunology ; HIV Infections ; drug therapy ; immunology ; Humans ; Interleukin-17 ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Helper-Inducer ; drug effects ; immunology ; Th1 Cells ; drug effects ; immunology ; Young Adult

OBJECTIVETo investigate the effects of dexamethasone on Th1/Th2 cytokines in oral lichen planus.

METHODSPeripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation from OLP patients and healthy controls. PBMC from patients with OLP were stimulated with phytohemagglutinin (PHA) and dexamethasone respectively for 72 h. The concentrations of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) in culture supernatants were determined by ELISA. The mRNA levels for IFN-gamma and IL-4 in culture cells were evaluated using semi-quantitative reverse transcription-polymerase chain reaction.

RESULTSCompared with healthy controls, the levels of IFN-gamma in OLP patients were significantly lower (P < 0.05). The levels of IL-4 were higher, but not statistically significant (P > 0.05). The ratios of IFN-gamma/IL-4 were lower in patients with OLP (P < 0.05). Dexamethasone significantly inhibited the levels of IFN-gamma and IL-4 (P < 0.01). Moreover, IFN-gamma was inhibited significantly more than IL-4. The levels of IFN-gamma and IL-4 mRNA expression in culture cells were consistent with protein production in supernatants.

CONCLUSIONSTh2 immune response is predominant in OLP. Dexamethasone is an immunosuppressant inhibiting Th1/Th2 cytokines.

Adult ; Cells, Cultured ; Dexamethasone ; pharmacology ; Female ; Glucocorticoids ; pharmacology ; Humans ; Interferon-gamma ; analysis ; Interleukin-4 ; analysis ; Lichen Planus, Oral ; immunology ; Male ; Middle Aged ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology

OBJECTIVE: To investigate the protective effect of Zengye Decoction (, ZYD) on the submandibular glands (SMGs) in nonobese diabetic (NOD) mice. METHODS: Twenty-seven female NOD mice were randomly equally divided into 3 groups: the model group, the hydroxychloroquine (HCQ) group, and the ZYD group. Nine C57/B6 mice served as the normal group. After 1-week acclimation, the HCQ and ZYD groups were intragastrically administered with HCQ and ZYD, respectively, and the normal and model groups were administered with normal saline. Changes in the salivary flow rate were observed. Mice from all 4 groups were sacrificed at the age of 20 weeks. The serum and SMGs were collected. Serum cytokines gamma-interferon (IFN-γ), interleukin-10 (IL-10) were detected by enzyme-linked immunosorbent assay. Histological changes in the submandibular glands were examined by hematoxylin and eosin staining. The mRNA expression of IFN-γ, IL-10 and vasoactive intestinal peptide (VIP) in the submandibular glands were measured by real-time polymerase chain reaction. RESULTS: Compared with the model group, the salivary flow of the ZYD group significantly increased (P<0.05), the extent of the histological changes was ameliorated (P<0.05), and the Th1/Th2 cytokine imbalance was remedied (P<0.05). In the ZYD-treated mice, the VIP mRNA was up-regulated (P<0.05). CONCLUSIONS ZYD is beneficial in protecting structure and function of SMGs in NOD mice. The mechanism may be associated with the correction of the Th1/Th2 cytokine imbalance, and with the prevention of a progressive decline of the VIP level.
Animals ; Cytokines ; blood ; Drugs, Chinese Herbal ; pharmacology ; Female ; Mice ; Mice, Inbred C57BL ; Mice, Inbred NOD ; Salivation ; drug effects ; Sjogren's Syndrome ; drug therapy ; immunology ; Submandibular Gland ; drug effects ; pathology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Vasoactive Intestinal Peptide ; genetics