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Aim: To create a method of detecting typeII natural killer T (NKT) cells of mice., Methods: Biotinylated mouse CD1d monomers were mixed with sulfatide at a molar ratio of 1:3 (protein:lipid) and incubated at room temperature overnight, and then 80 μg of streptavidin-PE was added into 200 μg of the CD1d-sulfatide mixture and incubated at room temperature for 4 h to get sulfatide/CD1d tetramer. Flow cytometry was used to detect the percentage of typeII NKT cells in mononuclear cells (MNCs) of lung and spleen of normal mice, as well as the percentage of typeII NKT cells in spleen MNCs of mice after stimulated with sulfatide., Results: In normal mice, the percentage of typeII NKT cells accounted for (0.875±0.096)% and (1.175±0.263)% in MNCs of spleen and lung; the percentage in spleen MNCs after activated with sulfatide was (2.75±0.603)%, which significantly increased as compared with that in normal mice (P<0.01)., Conclusion: Sulfatide-loaded CD1d tetramer is an effective method of detecting typeII NKT cells in mice.

Adult-onset Still disease (AOSD) is an uncommon inflammatory condition of unknown origin and pathogenesis. Pulmonary involvement is rare and includes pleuritis and transient radiological infiltrations. We report two cases of AOSD characterized by lung involvement at presentation. Both were misdiagnosed as pneumonia with para-pneumo­nic effusion. We also discuss the difficulties in diag­nosis of AOSD with pulmonary infiltration.

Objective: To investigate the deficiency of the expression of the phenotypes (CD(1a), CD(83), CD(40), CD(86)) and cytokines (IL-12 and IL-10) by human peripheral blood monocyte (PBMC)-derived dendritic cell (DCs) from asthmatic subjects, and their influence on naive T cell polarization., Methods: Adherent cells were isolated from peripheral blood samples in asthmatic patients and in healthy volunteers, and were cultured with granulocyte-macrophage colony-stimulating factor and IL-4 as immature DC (iDC). iDCs were stimulated with lipopolysaccharide as mature DC (mDCs). Nonadherent cells were obtained from umbilical cord blood by idem methods, and naïve T cells were sorted by adding anti-CD(4) and anti-CD(45RA) in nonadherent cells respectively and magnetic microbeads. Naïve T cells and mDCs from two groups were co-cultured in complete RPMI1640 media respectively, and naive T cells polarized as T helper cells 1 (Th1) and Th2. The expression of the CD(1a), CD(83), CD(40) and CD(86) on mature DCs were examined by fluorescent activated cell sorter. IL-12 and IL-10 released by mDCs and IL-4 and IFNgamma produced by Th cells were measured by ELISA., Results: (1) The expression of CD(86) on dendritic cells from atopic asthmatics was higher than that from healthy control subjects (40.75 +/- 3.99 vs 29.88 +/- 1.25, P < 0.01). (2) The levels of IL-12, IL-12p40 and IL-10 produced by DCs from asthmatic subjects were all significantly lower than those from healthy control group (217.79 +/- 118.65 vs 905.66 +/- 495.32, P < 0.01; 2072.22 +/- 1496.37 vs 5569.43 +/- 2922.75, P < 0.01; 336.89 +/- 261.52 vs 1425.00 +/- 1148.87, P < 0.05, respectively). (3) IL-4 production by Th2 cells which were primed by DCs from asthmatics was significantly increased as compared to that from control group (368.56 +/- 190.72 vs 584.91 +/- 290.13, P < 0.01); On the contrary, IFNgamma in the patient group was reduced as compared to that in the control group (425.33 +/- 164.94 vs 49.86 +/- 18.14, P < 0.05). (4) In the patient group, the level of IL-12 was positively correlated to that of IFNgamma (P < 0.05), negatively correlated to that of IL-4 (P < 0.05); IL-10 was negatively correlated to IL-4 (P < 0.05). (5) There was a positive correlation between IL-12 and IL-10 in the two groups (P < 0.01)., Conclusion: Because of DC deficiency, naïve T cells preferentially polarize to Th2 which synthesize more Th2-type cytokine (i.e. IL-4) and T cell tolerance cannot be induced, which may be one of the important pathogenic mechanisms for allergic asthma.

Aim: To investigate the effect of theophylline on the naive T cell differentiation and the probable role of interleukin-12 (IL-12)., Methods: Naive cord blood T cells were treated with theophylline 10 mg/L for 3 d after stimulation with PHA 100 mg/L. Differentiation of T cells was analyzed by flow cytometry. Theophylline 10 mg/L and IL-12-mAb 0.025 mg/L were added in cord blood mononuclear cell (CBMC) cultures primed with LPS 1 mg/L to detect the levels of IL-12 and IL-12P40. The whole blood cultures were obtained from twelve health volunteers with or without administration of theophylline (200 mg). Cytokines were measured by enzyme linked immuno-sorbent assay., Results: Theophylline promoted T helper 1 (Th1) cells differentiation from naive T cells (21.9 %+/-10.3 % vs 9.4 %+/-5.6 %, P<0.05), but had no significant effect on Th2 deviation. But theophylline inhibited the production of IL-12 and IL-12P40 by CBMC in vitro (28+/-6 ng/L vs 57+/-14 ng/L and 88+/-34 ng/L vs 214+/-82 ng/L, P<0.01) and reduced IL-12 and IL-12P40 levels in whole blood cultures from healthy subjects (19+/-11 ng/L vs 31+/-15 ng/L and 92+/-13 ng/L vs 196+/-49 ng/L, P<0.01)., Conclusion: Theophylline promoted the differentiation of Th1 cells. IL-12 seemed not to be involved in this process.

Aim: To investigate the mechanism of anti-inflammatory action of theophylline on asthma., Methods: Nineteen asthmatic subjects were administered 200 mg sustained-release theophylline twice daily for 4 weeks. The number of CD4+ T lymphocytes, eosinophils, and the levels of interleukin (IL)-5 and interferon gamma (IFN-gamma) in sputum pre- and post-administration were measured by direct immunofluorescene technique, Wright's stain and enzyme-linked immunosorbent assay, respectively. The symptom scores and lung function were also evaluated., Results: Theophylline treatment significantly improved symptom scores and FEV1.0, FEV1.0 % (P < 0.05) and reduced sputum eosinophils (P < 0.01) in asthmatic subjects. These were accompanied by a decrease in sputum IL-5 level (P < 0.01), but sputum CD4+ T lymphocytes and IFN-gamma had no significant change (P >0.05). The mean (range) serum theophylline concentration in final steady state was 7.9 (3.9 - 12.9) mg/L., Conclusion: The anti-inflammatory action of theophylline in asthma may result from reduction of IL-5 production in the airways.