OBJECTIVETo explore the characteristics of B cell-activating factor (BAFF) secreted by peripheral blood monocyte-derived dendritic cell (MoDC) in chronic idiopathic thrombocytopenic purpura (cITP) and the function of MoDC on B cell proliferation.

METHODSTen cITP patients were studied dynamically before and after treatment. The BAFF levels in serum and the supernatant of LPS stimulated MoDC were tested with ELISA. The BAFF gene expression in LPS stimulated MoDC was tested with RQ-PCR, the B cell proliferation co-cultured with the supernatant of LPS stimulated MoDC for 5 days was tested with flow cytometry for CFSE and (3)H thymidine incorporation.

RESULTSThe BAFF level in serum (serum BAFF) \[(2461 ± 483) ng/L\], and supernatant of LPS stimulated MoDC (supernatant BAFF) \[(1113 ± 113) ng/L\] and BAFF mRNA in LPS stimulated MoDC (BAFF mRNA) (1.70 ± 0.23) before treatment were higher than that after treatment \[(621 ± 53) ng/L, (490 ± 49) ng/L and 0.37 ± 0.12\] and normal group \[(742 ± 77) ng/L, (582 ± 63) ng/L and 0.52 ± 0.08\]. There was a positive correlation among serum BAFF, supernatant BAFF and BAFF mRNA, and a negative correlation among serum BAFF, supernatant BAFF and BAFF mRNA and blood platelet count (BPC) in all ITP patients. The supernatant of LPS-stimulated MoDC from untreated patients enhanced B cell proliferation as compared with the supernatant of LPS-stimulated MoDC from treated patients and normal group.

CONCLUSIONBAFF might contribute to disease development in cITP. MoDC may directly increase B cell proliferation by secreting BAFF without T cell help, playing an important role in the antibody production in cITP.

B-Lymphocytes ; immunology ; Dendritic Cells ; immunology ; Humans ; Interleukin-4 ; Monocytes ; Purpura, Thrombocytopenic, Idiopathic ; immunology

OBJECTIVETo investigate the influence of the LPS of Bacteroides fragilis on the secretion of IL-2 and IL-4 from the peripheral blood mononuclear cells of normal individuals, so as to elucidate the mechanism of the infection by Bacteroides fragilis.

METHODSLPS was obtained from both the strains isolated from patients and from standard NCTC9343. Peripheral blood mononuclear cells (PBMCs) were treated with different concentrations of LPS thus obtained. The supernatants from the cell culture of the PBMCs were harvested at 24 PBHs and were subjected to the determination of the IL-2 and IL-4 contents by ELISA method. RESULTS The IL-2 secretion from the PBMCs of normal volunteers was obviously inhibited by the LPS from Bacteroides fragilis (P < 0.01), and the inhibitory effect was dose-dependent. Nevertheless, the IL-4 secretion from the PBMCs of normal volunteers was significantly stimulated by the LPS from Bacteroides Fragilis (P < 0.05), and it was not concentration dependent. There was no difference between the effects of the LPSs from patients and standard strains (P < 0.05).

CONCLUSIONThe LPS from Bacteroides fragilis was inhibitory to the secretion of IL-2 from PBMCs and was stimulative to that of IL-4 from PBMCs of normal human persons.

Bacteroides fragilis ; metabolism ; Cells, Cultured ; Humans ; Interleukin-2 ; immunology ; secretion ; Interleukin-4 ; immunology ; secretion ; Lipopolysaccharides ; pharmacology ; Monocytes ; drug effects ; immunology

Spontaneous remission (SR) of leukemia is a rare event in clinic, which possibly correlated with severe infection and sepsis, but its exact mechanism has not been confirmed. Plasmacytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) play a key role in innate and adaptive immunity respectively. A patient with severe infection of staphylococcus aureus acquired completely spontaneous remission (SR), moreover a increased number of pDC were observed, suggesting that bacteria-activated pDC may play an important role in SR. This study was purposed to explore if the bacteria can stimulate pDC successfully and get a functional pDC. Both pDC and mDC were isolated from freshly collected, leukocyte-rich buffy coats from healthy blood donor and leukemic patient with SR by using MACS and FACS. The pDC were cultured in RPMI 1640 medium and were stimulated with different kinds of bacteria and the expression of CD40, CD86 and HLA-DR on the cell surface was analyzed by flow cytometry. The cytokine (IFN-α, IL-12, IFN-γ, IL-2, IL-4, IL-10) production was measured by using ELISA kits. The results showed that the stimulation with staphylococcus aureus and pseudomonas aeruginosa resulted in the maturation of pDC, which secrete a large number of IFN-α and promote the differentiation of naive CD4⁺ T cells to Th1 cells. The activated pDC expressed high level of CD40 and CD86 and showed higher T cell stimulatory capacities. It is concluded that staphylococcus aureus and pseudomonas aeruginosa can activate pDC, the activated pDC secrete high quantity of IFN-α. This result suggests that bacteria stimulated pDC may play a key role in SR of leukemia following severe infections.
CD4-Positive T-Lymphocytes ; Dendritic Cells ; immunology ; microbiology ; Humans ; Interferon-alpha ; Interleukin-10 ; Interleukin-12 ; Interleukin-2 ; Interleukin-4 ; Leukemia ; diagnosis ; immunology ; microbiology ; Remission, Spontaneous ; Staphylococcus aureus

OBJECTIVETo determine the proportion of Tc17 cells in the lungs of mice with neutrophilic(NEU) asthma, and to investigate the role of Tc17 cells in the pathogenesis of NEU asthma.

METHODSThirty-two C57/B6 mice of clean grade were randomly divided into two groups: NEU asthma and control. The mice in the NEU asthma group were sensitized by airway instillation of ovalbumin (OVA) and lipopolysaccharide (LPS), and challenged with an aerosol of OVA, while those in the control group were sensitized and challenged with normal saline. At 24 hours after the final challenge, bronchoalveolar lavage fluid (BALF) was collected, and the total number and differential counts of nucleated cells and percentage of each type were determined. The lung tissues were separated and hematoxylin-eosin staining was performed to observe the pathological changes of lungs; flow cytometry was applied to determine the percentages of Tc17 and Th17 cells in the lung tissues. Enzyme-linked immunosorbent assay was applied to determine the levels of interleukin-6 (IL-6), transforming growth factor β (TGF-β), and interleukin-17 (IL-17) in BALF.

RESULTSThe NEU asthma group had a significantly higher total number of nucleated cells, a significantly higher percentage of eosinophils, and a significantly higher percentage of neutrophils in BALF than the control group (P<0.01). The NEU asthma group also had significantly higher percentages of Tc17 and Th17 cells than the control group (P<0.01). In the NEU asthma group, the percentage of Tc17 cells was positively correlated with that of Th17 cells (P<0.05). The NEU asthma group had significantly higher concentrations of IL-6, TGF-β, and IL-17 in BALF than the control group (P<0.05).

CONCLUSIONSThe expression of Tc17 cells in the lung tissues increases in mice with NEU asthma, and the increased number of Tc17 cells may be involved in the pathogenesis of NEU asthma. Tc17 cells may play an important role in NEU asthma through IL-17.

Animals ; Asthma ; genetics ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Interferon-gamma ; immunology ; Interleukin-17 ; genetics ; immunology ; Interleukin-4 ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Lymphocyte Count ; Mice ; Th17 Cells ; immunology

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

The present study aimed to examine the effect of interleukin (IL)-4 on neutrophil chemotaxis in airway inflammation in asthmatic rats and the possible mechanism. Male Wistar rats were intranasally instilled with recombinant rat (rr) IL-4 (rrIL-4) at different doses [2, 4 or 8 μg/animal, dissolved in 200 μL normal saline (NS)] or rrIL-4 at 4 μg/animal (dissolved in 200 μL NS). NS (200 μL) and LPS (6 mg/kg/animal, dissolved in 200 μL NS) were intranasally given respectively in the negative and positive control groups. Moreover, the asthmatic lung inflammation was induced in rats which were then intranasally treated with rrIL-4 (4 μg/animal) or LPS (6 mg/kg/animal). The normal rats treated with different doses of rrIL-4 and those asthmatic rats were sacrificed 6 h later. And animals instilled with rrIL-4 at 4 μg were sacrificed 6, 12 or 24 h later. The bronchoalveolar lavage fluid (BALF) and lungs were harvested for detection of leukocyte counts by Wright-Giemsa staining and lung histopathology by haematoxylin-eosin (HE) staining. The levels of cytokine-induced neutrophil chemoattractant (CINC)-1 and intercellular adhesion molecule (ICAM)-1 in BALF were determined by ELISA. Real-time PCR was used to measure the mRNA expression of CINCs (CINC-1, CINC-2α, CINC-2β, CINC-3) and ICAM-1 in lung tissues. The results showed that the intranasal instillation of IL-4 did not induce a recruitment of neutrophils in BALF in rats. However, IL-4 could increase the CINC-1 level in BALF in a dose-dependent manner at 6 h. But the mRNA expression levels of CINC-1, CINC-2α, CINC-2β, CINC-3 were not significantly increased in lungs of IL-4-treated rats relative to NS negative control group. Moreover, IL-4 was found to augment the mRNA expression of ICAM-1 in lungs and the ICAM-1 level in BALF at 6 h. However, the increase in CINC-1 and ICAM-1 levels in BALF of IL-4-treated asthmatic rats was not significantly different from that in untreated asthmatic rats. These findings indicate that IL-4 does not directly recruit neutrophils in the rat lungs, but it may contribute to airway neutrophilia through up-regulation of CINC-1 and ICAM-1.
Animals ; Asthma ; immunology ; Chemotactic Factors ; immunology ; Intercellular Adhesion Molecule-1 ; immunology ; Interleukin-4 ; immunology ; Lung ; immunology ; Male ; Neutrophils ; immunology ; Rats ; Rats, Wistar

OBJECTIVE: To obtain a global view of lymphocyte subset changes in the peripheral blood and cytokine profile in patients with class IV lupus nephritis (LN). METHODS: A total of 30 patients with biopsy proven active class IV LN, 30 patients with biopsy proven active class V LN, and 30 healthy controls were enrolled. Serum concentration of Th1 cytokines (IFN-γ, IL-1, IL-2, and TNF-α) and Th2 cytokines (IL-4, IL-5, IL-6, IL-10, IL-13) were simultaneously analyzed by Fast Quant Human Th1/Th2 protein array. The expression of lymphocyte subsets was measured by flow cytometer. Clinical parameters such as urine protein of 24 h, autoantibodies and complement were detected. Pearson analysis was used to examine the relation between lymphocyte subsets and clinical parameters, cytokine and clinical parameters. RESULTS: The patients with class IV LN had evident anemia (P<0.001), hypocomplementemia, and hypoalbuminemia (P<0.05). There were no significant difference both in the ratio and number of CD4+ lymphocytes between the controls and the patients. In the patients with class IV LN, the ratio and number of CD4+ lymphocytes were both lower than those of the controls (P<0.01). The ratio and number of CD20+ lymphocytes were both higher than those of the controls (P<0.05), and a significant decrease in CD4+CD25+Foxp3+ regulatory T cells (Tregs) was observed in the patients compared with healthy age-matched controls (P<0.001). The abnormality of lymphocytes in class IV patients was obviously notable, especially in CD4+CD25+Foxp3+ regulatory T cells. In class IV patients, most of the detected cytokines levels were markedly elevated as compared with the controls, including Th2 cytokines INF-γ (P<0.05), IL-2 (P<0.05) and TNF-α (P<0.01), and Th2 cytokines IL-4 (P<0.05), IL-6 (P<0.05), IL-10 (P<0.01) and IL-13 (P<0.01). Only 4 out of 9 cytokines significantly increased in class V patients. In addition to IL-2, all of them belonged to Th2 (IL-4, IL-10 and IL-13) cytokines. There was negative correlation between CD4+CD25+Foxp3+ regulatory cells and urine protein, anti-dsDNA titer or SLEDAI (r=-0.781, -0.746 and -0.646, respectively; P<0.05). There was positive correlation between IL-5 and anti-dsDNA titer (r=0.708, P<0.05), between IL-5 and creatinine (r=0.681, P<0.05), and between IL-10 and SLEDAI (r=0.877, P<0.01). There was also negative correlation between IL-10 and urine protein of 24 h (r=-0.659, P<0.05), between IL-10 and hemoglobin concentration (r=-0.856, P<0.01), and between IL-13 and urine protein of 24 h (r=-0.769, P<0.05). There was little correlation between cytokines and clinical parameters in patients with class V LN. CONCLUSION There is extensive abnormality in lymphocyte subsets and cytokine profile in patients with class IV LN, which may be the mechanism of immunosuppressive agents to treat patients with class IV LN.
Cytokines ; immunology ; Flow Cytometry ; Humans ; Interleukin-1 ; Interleukin-10 ; Interleukin-2 ; Interleukin-4 ; Interleukin-5 ; Interleukin-6 ; Lupus Nephritis ; classification ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Tumor Necrosis Factor-alpha

Animals ; Asthma ; immunology ; Diabetes Mellitus, Experimental ; immunology ; Interferon-gamma ; biosynthesis ; Interleukin-4 ; biosynthesis ; Male ; Ovalbumin ; immunology ; Rats ; Rats, Sprague-Dawley ; Streptozocin ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVE: To explore the relationship of the expression of thymic stromal lymphopoietin (TSLP) in nasal polyps tissues and the Th2 inflammatory response. METHOD: Sixty patients with nasal polyps were collect ed. The immunohistochemical staining method was used to detect the expression of TSLP in nasal polyps tissues and ELISA method to the expression of IL-4, IL-5, IFN-gamma, IL-13 and analyzed the correlation between them. RESULT: The expression of TSLP in nasal polyp tissues was higher than that in normal inferior turbinate mucosa (P < 0.05). The expression level of IL-4, IL-5, IFN-gamma and IL-13 in nasal polyps tissues were significantly higher than those in control group (P < 0.05). TSLP staining was a statistically significant positive correlation with IL-4, IL5 and IL-13 (r = 0.475, 0.594 and 0.582, respectively, P < 0.01), while inverse correlation with IFN-gamma (r = -0.614, P < 0.01). CONCLUSION The high expression of TSLP might promote T cell differentiation towards Th2, and participated in the occurrence/development of nasal polyps, aggravated the nose Th2 inflammatory response.
Adult ; Cytokines ; metabolism ; Female ; Humans ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-5 ; metabolism ; Male ; Nasal Polyps ; immunology ; metabolism ; Th2 Cells ; immunology ; Young Adult

OBJECTIVETo study the concentrations of IL-4 and IL-13 in bronchoalveolar lavage fluid (BALF) in neonates with respiratory distress syndrome (RDS) and concurrent ventilator-associated pneumonia (VAP).

METHODSSixty-eight neonates with RDS undergoing mechanical ventilation for over 48 hrs were enrolled. IL-4 and IL-13 levels in BALF were measured using ELISA 1, 72 and 96 hrs after mechanical ventilation. The results were compared between the neonates with concurrent VAP (n=37) and without (n=31).

RESULTSThe levels of BALF IL-4 96 hrs after ventilation in the VAP group (35.34+/-1.78 ng/mL) were significantly higher than those in the non-VAP group (13.69+/-2.47 ng/mL, P<0.05). The levels of BALF IL-13 96 hrs after ventilation in the VAP group (33.74+/-2.74 ng/mL) also increased significantly compared with those in the non-VAP group (13.50+/-3.81 ng/mL) (P<0.05). There were significant differences in BALF IL-4 and IL-13 levels between 1 hr and 96 hrs in the VAP group (P<0.05).

CONCLUSIONSBALF IL-4 and IL-13 levels increase in neonates with RDS and concurrent VAP. IL-4 and IL-13 may involve in the regulation of the inflammatory immune response.

Bronchoalveolar Lavage Fluid ; immunology ; Female ; Humans ; Infant, Newborn ; Interleukin-13 ; analysis ; Interleukin-4 ; analysis ; Male ; Microbial Sensitivity Tests ; Pneumonia, Ventilator-Associated ; immunology ; microbiology ; Respiratory Distress Syndrome, Adult ; immunology