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Objective: to investigate the levels of peripheral CD(8)(+)CD(25)(+)Foxp(3)(+) regulatory T cells (CD(8)(+)CD(25)(+)Treg) in stable chronic obstructive pulmonary disease (COPD) patients, and the effect of muscarinic cholinergic receptor antagonist, tiotropium bromide, on the expression of CD(8)(+)CD(25)(+)Treg., Methods: from Oct. 2007 to Mar. 2008, 23 patients with stable moderate to severe COPD received tiotropium bromide inhalation (18 microg daily) for 3 months. Before and after the use of tiotropium bromide, lung function was performed and peripheral vein blood samples were collected from the patient. Lymphocytes were isolated by three-color labeled monoclonal antibodies flow cytometry to detect the quantity and percentage of CD(8)(+)T cell, CD(8)(+)CD(25)(+) T cell, CD(8)(+)CD(25)(+)Treg, CD(4)(+)T cell, CD(4)(+)CD(25)(+) T cell and CD(4)(+)CD(25)(+)Treg, respectively. Paired t test was used for comparison between data before and after treatment., Results: in patients with stable COPD, after tiotropium bromide treatment, the percentage of CD(4)(+) T cells was increased from (28 +/- 10)% to (36 +/- 6)%, and the difference was significant (t = 3.20, P < 0.01). CD(4)(+)CD(25)(+) was decreased from (10 +/- 7)% to (4 +/- 3)% (t = 3.78, P < 0.01), and CD(8)(+)CD(25)(+)Treg was increased from (8 +/- 8)% to (21 +/- 21)% (t = 2.72, P < 0.05). At baseline, CD(8)(+) cells, CD(8)(+)CD(25)(+) cells and CD(4)(+)CD(25)(+)Treg were detectable in the peripheral blood, but no significant changes were observed after treatment. After treatment with tiotropium bromide, the lung function was markedly improved; FEV(1), FEV(1)/Pre%, and FEV(1)/FVC were increased from (1.0 +/- 0.3) L, (35 +/- 10)% and (41 +/- 8)% to (1.1 +/- 0.3) L, (40 +/- 11)% and (45 +/- 11)%, respectively (t = 2.65, 2.56 and 2.37, respectively, all P < 0.05). By linear correlation analysis, the quantity of changes of CD(4)(+) T cells and CD(4)(+)CD(25)(+) T cells were negatively correlated with the rate of change in CD(8)(+)CD(25)(+)Treg (r = -0.61, P < 0.05 and r = -0.72, P < 0.01, respectively)., Conclusions: in the peripheral blood of patients with stable COPD, there was expression of CD(8)(+)CD(25)(+)Treg and CD(4)(+)CD(25)(+)Treg. The muscarinic receptor antagonist tiotropium bromide, promoted the amplification of CD(4)(+), inhibited the expression of CD(25)(+), and enhanced the expression of CD(8)(+)Treg, suggesting that muscarinic receptor antagonist tiotropium bromide may possess immunomodulation function.

Objective: To investigate whether house dust mite (HDM) could induce CD(4)(+) CD(25)(+) T cells infiltration into asthmatic airways in patients vivo., Methods: Ten subjects with asthma underwent initial bronchoscopy during which normal saline and HDM were administered to two sublobar segments separately. The second bronchoscopy were carried out and bronchoal lavage fluid from HDM-challenged sites and saline-challenged sites were separately taken 24 h later. The differential cell counts were determined, and the absolute number of each type was calculated. At the same time, CD(3)(+), CD(4)(+) and CD(4)(+) CD(25)(+) T cells were determined by flow-cytometric analysis. We compared cellular counts in airways without and after topical instillation of HDM., Results: Eosinophile granulocyte cells of broncho-alveolar fluid in the HDM-challenged sites (1.4 +/- 0.1) x 10(6)/ml are more than it in control sites (0.3 +/- 0.1) x 10(6)/ml, P < 0.003. Lymphocyte cells of BALF in the HDM-challenged sites (2.2 +/- 0.3) x 10(6)/ml are more than it in control sites (0.3 +/- 0.1) x 10(6)/ml, P < 0.001; CD(4)(+) CD(25)(+)T cells of BALF in the HDM-challenged sites (784.0 +/- 281.3) cell/microl are more than it in control sites (7.7 +/- 3.6) cell/microl, P < 0.001., Conclusions: Our findings suggest that HDM is capable of inducing CD(4)(+) CD(25)(+) T cells recruitment into non-acute mild allergic asthmatic airways.

Objective: To explore whether regulatory CD(4)(+)CD(25)(+) cells exist in patients with atopic asthma., Methods: The numbers of peripheral blood CD(4)(+)CD(25)(+) cells in peripheral blood of atopic asthmatics and healthy nonatopic subjects were determined using flow cytometry. CD(4)(+)CD(25)(+) and CD(4)(+)CD(25)(-) cells from atopic asthmatics and normal donors were isolated, and were cultured to observe the effects of CD(4)(+)CD(25)(+) cells on proliferation response as well as Th1/Th2 cytokine production of CD(4)(+)CD(25)(-) cells in vitro., Results: A significant increase in CD(4)(+)CD(25)(+) cell numbers was shown in atopic asthmatic patients during acute exacerbation [(14.9 +/- 1.8)%, P < 0.01], but not in patients with stable asthma [(11.8 +/- 0.7)%] and normal subjects [(11.2 +/- 0.8)%, P > 0.05]. The mean inhibition values of the proliferation response of CD(4)(+)CD(25)(-) cells by CD(4)(+)CD(25)(+) cells from normal controls [(72 +/- 8)%] and asthmatics [(74 +/- 9)%] were similar (P > 0.05). There was no difference in inhibitory effects on both Th1 and Th2 cytokine production of CD(4)(+)CD(25)(-) cells by CD(4)(+)CD(25)(+) cells in the two groups., Conclusion: Although CD(4)(+)CD(25)(+) cells increase in atopic asthma during exacerbation, these regulatory T cells appear to function normally with regard to their suppressive activities.

Objective: Interleukin-16 (IL-16) is a chemoattractant of CD4+ lymphocytes, and it has been implicated in the pathogenesis of various inflammatory diseases. The aim of the present study was to measure IL-16 in pleural effusions caused by tuberculosis and malignancy and its relationship with cell and differential counts as well as lymphocyte subsets., Methods: Pleural effusion and venous blood samples were collected from 32 patients with tuberculous pleuritis and 30 lung cancer patients with malignant effusion. Analysis of pleural effusion for total leukocytes and cell differentials of leukocytes was performed. Three-color flow cytometry was performed to determine T lymphocyte subsets in cell pellets of pleural effusion. The concentration of IL-16 in cell-free supernatants of pleural effusion and sera was measured by a sandwich enzyme-linked immunosorbent assay., Results: In all the studied patients, the level of IL-16 was significantly higher in pleural effusion than in serum. The levels of IL-16 were significantly higher in tuberculous than in malignant effusions. In pleural effusion, positive correlations were found between the IL-16 levels and total cell counts, lymphocytes, CD3+ T cells, as well as CD4+ T cells., Conclusions: Compared to malignant pleural effusion, IL-16 appeared to be increased in tuberculous pleural effusion. The pleural effusion IL-16 levels were positively related to the numbers of CD4+ T cells, suggesting that IL-16 might be capable of inducing CD4+ T cell infiltration into pleural space.

Objective: To explore the mode of immune responses of gammadeltaT cells in asthma and the role of gammadeltaT cell subsets in the pathogenesis of asthma., Methods: Wistar rats were sensitized and challenged with ovalbumin to establish an asthmatic model (n = 10, for each group). "Attack-panning" method was used to culture selective gammadeltaT cells. The percentage of gammadelta T cells among cultured cells was determined by flow cytometry. The levels of interleukin (IL)-4 and interferon-gamma (IFN-gamma) were measured by ELISA. The expression of IL-4 mRNA and IFN-gamma mRNA was determined by in situ hybridization., Results: In the asthmatic group, IL-4 and IFN-gamma were detectable at the same time in cultured supernatant of gammadeltaT cells in peripheral blood mononuclear cells (PBMC) or broncho-alveolar fluid (BALF), and the level of IL-4 [PBMC: (40.5 +/- 3.7) ng/L, BALF: (49.6 +/- 3.1) ng/L] increased as compared with the control group [PBMC: (26.1 +/- 2.1) ng/L, BALF: (23.6 +/- 1.7) ng/L; all P < 0.01], while the level of IFN-gamma [PBMC: (17.6 +/- 2.5) ng/L, BALF: (28.5 +/- 3.6) ng/L] decreased as compared with the control group [PBMC: (24.3 +/- 1.7) ng/L, BALF: (38.4 +/- 2.8) ng/L; all P < 0.01]. In the asthmatic group, the percentage of IL-4 mRNA positive-staining cells [PBMC: (76.2 +/- 7.2)%, BALF: (85.7 +/- 8.4)%] among the gammadeltaT cells significantly increased as compared with the control group [PBMC: (20.6 +/- 5.3)%, BALF: (25.0 +/- 6.8)%; all P < 0.01], while the percentage of IFN-gammamRNA positive-staining cells [PBMC: (30.9 +/- 6.7)%, BALF: (41.5 +/- 3.6)%] among the gammadeltaT cells significantly decrease as compared with the control group [PBMC: (60.1 +/- 4.2)%, BALF: (53.8 +/- 5.1)%; all P < 0.01]., Conclusions: gammadeltaT cells can secrete both IL-4 and IFN-gamma at the same time, or there are two subsets of gammadeltaT cells which can secrete either IL-4 or IFN-gamma in PBMC or BALF in asthma. Th1/Th2 immune response mode is present in gammadeltaT cells of asthma and Th2-cytokine profile predominates. gammadeltaT cells is involved in the pathogenesis of asthma.

Objective: The aim of the present study was to evaluate whether eosinophils within the tracheobronchial lumen can stimulate Th2 cell expansion by presenting antigen both in vitro and in vivo., Methods: Airway eosinophils were recovered from ovalbumin-sensitized and -challenged BALB/c mice, these eosinophils were then cocultured with sensitized CD4(+) cells in the absence or presence of anti-CD80 or/and -CD86 monoclonal antibodies. Airway eosinophils were instilled into the trachea of sensitized mice, At 3 d thereafter, the draining paratracheal lymph nodes were removed and teased into cell suspensions for culture. Cell-free culture supernatants were collected for detection of cytokines., Results: Our data showed that airway eosinophils, recovered following inhalational ovalbumin challenge in sensitized mice functioned as CD80- and CD86-dependent antigen-presenting cells to stimulate sensitized CD4(+) lymphocytes to produce IL-4, IL-5 and IL-13, but not interferon-gamma (IFN-gamma) in vitro assay. When instilled intratracheally in ovalbumin-sensitized recipient mice, these antigen-loaded eosinophils migrated into draining paratracheal lymph nodes primed Th2 cells in vivo for IL-4, IL-5 and IL-13, but not IFN-gamma, production during the in vitro culture that was also CD80- and CD86-dependent., Conclusions: We concluded that eosinophils within the lumina of airways could process inhaled antigen function in vitro and in vivo as antigen-presenting cells to promote expansion of Th2 cells. This investigation highlights the potential of eosinophils to not only act as terminal effector cells but also to actively modulate immune responses by amplifying Th2 cell responses.

Objective: To study the pathological features of airway inflammation and remodeling in rats with chronic bronchitis (CB) and emphysema and to evaluate the protective and therapeutic effects of erythromycin (EM)., Methods: Forty-three Wistar rats were assigned to eight groups: normal control group (A group, n = 5), normal saline solution group (P group, n = 5), CB group (L group, n = 6), CB and emphysema group (S group, n = 6), low-dose EM-treatment group (E(1) group, n = 5), high-dose EM-treatment group (E(2) group, n = 6), low-dose EM-prevention group (E(10) group, n = 5) and high-dose EM-prevention group (E(20) group, n = 5). The rat model of CB and emphysema was established by intratracheal instillation of lipopolysaccharide (LPS) and daily exposure to cigarette smog. After four weeks, total and differential cell counts in bronchoalveolar lavage fluid (BALF) were observed, and the pathomorphological changes in the lung were analyzed. The thickness of the smooth muscles and collagen in the bronchial wall were measured. Expression and localization of transforming growth factor beta(1) (TGF-beta(1)) were observed in the bronchi and lung tissues by immunohistochemistry. The levels of hyaluronic acid (HA) and procollagen type III (PCIII) in the serum and BALF were determined by the radioimmunoassay (RIA)., Results: (1) Compared with A group [(0.9 +/- 0.7) x 10(5)/ml], absolute neutrophil count in BALF from S group [(17.1 +/- 10.8) x 10(5)/ml] were significantly higher (P < 0.01). (2) Both the pathologic scores obtained from the S group (329 +/- 114) and P group (67 +/- 25), and the thickness of smooth muscles and collagen from S group [(9.6 +/- 2.6)%] and A group [(6.1 +/- 1.8)%] were statistically different (P < 0.01, P < 0.05, respectively). Expression of TGF-beta(1) in the lung of S group was significantly higher than that in A group. (3) The levels of HA [(152.5 +/- 36.3) micro g/ml] and PCIII [(40 +/- 8) micro g/ml] in serum and the levels of HA [(94 +/- 35) micro g/ml] and PCIII [(39 +/- 7) micro g/ml] in BALF in S group were higher than those in A group (P < 0.01). (4) After treatment with 100 mg/kg EM, absolute neutrophil count in BALF, the pathologic scores, the thickness of smooth muscles and collagen in the bronchi, the levels of PCIII and HA in serum and the levels of PCIII and HA in BALF were reduced to (2.1 +/- 1.4) x 10(5)/ml, 187 +/- 61, (6.0 +/- 2.3)%, (9.69 +/- 5.61) micro g/ml, (63.0 +/- 11.6) micro g/ml, (16 +/- 6) micro g/ml, (52 +/- 12) micro g/ml, respectively. Statistical analysis revealed that there were significant differences as compared to those of group S (P < 0.05)., Conclusions: Many inflammatory cells especially neutrophils and alveolar macrophages might play an important role in the airway inflammation of CB and emphysema. Thickening of smooth muscles and collagen in the bronchi and the excessive depositions of extracellular matrix (ECM) constitute the fundamental pathological characteristic of airway remodeling in CB and emphysema. EM may prevent airway inflammation and remodeling to some degree.