Effects of Medication Use on Small Airway Function and Airway Inflammation in Patients with Clinically Controlled Asthma.

OBJECTIVE: To observe effects of medication use on small airway function, airway inflammation and acute exacerbations in patients with clinically controlled asthma. METHODS: Forced expiratory flow over the middle half of the forced expiratory curve (FEF25%-75%), percentage of eosinophil, concentrations of eosinophil cationic protein (ECP) and interleukin (IL)-5 in induced sputum were assessed in patients with clinically controlled asthma who were given oral anti-inflammatory agents alone or in combination with inhaled therapy and inhaled therapy alone. Subsequently, acute exacerbations were compared between two groups during the 24-week follow-up period. RESULTS: FEF25%-75% in 43 patients with clinically controlled asthma given oral anti-inflammatory agents alone or in combination with inhaled therapy was significantly higher than that in 49 patients given inhaled therapy alone. Meanwhile, the percentage of eosinophils and levels of IL-5 and ECP in patients with clinically controlled asthma given oral anti-inflammatory agents alone or in combination with inhaled therapy were significantly lower than those in patients given inhaled therapy alone. Additionally, the patients with clinically controlled asthma given inhaled therapy were likely to have more acute exacerbation than the patients given oral anti-inflammatory agents alone or in combination with inhaled therapy during the 24-week follow-up period. CONCLUSION: Systemic anti-inflammatory agents may have a greater effect on parameters reflecting small airway patency and reducing acute exacerbations, presumably secondary to reduction in airway inflammation.

The Clinical Role of Changes of Maximum Expiratory Flow at 25% and 50% of Vital Capacity before and after Bronchodilator Reversibility Test in Diagnosing Asthma.

Changes of maximum expiratory flow at 25% and 50% of vital capacity (MEF25 and MEF50, respectively), and predominant parameters indicating small airways function in asthmatics before and after bronchodilator (BD) reversibility test have been less interpreted. Our study aimed to investigate the clinical role of changes of MEF25 and MEF50 before and after BD reversibility test in diagnosing asthma. Forced expiratory volume in the first second (FEV1), MEF25, and MEF50 were measured before and after BD reversibility test in 207 asthmatic patients using standard process. Forty healthy individuals were enrolled as controls. Receiver operating characteristic (ROC) curve was used to assess the diagnostic accuracy of reversibility of MEF25 and MEF50 before and after BD reversibility test (ΔMEF25% and ΔMEF50%, respectively) in diagnosing asthma. Among these functional criteria, ΔMEF25% and ΔMEF50% ≥ 25% performed the best diagnostic performance. The sensitivity, specificity, and accuracy of ΔMEF25% ≥ 25% as an objective diagnostic test for asthma were 63.29%, 87.50%, and 67.21%, and of ΔMEF50% ≥ 25% were 79.23%, 85.00%, and 80.16%, respectively. The area under the ROC curve of the indicators was 0.8203 and 0.9104, respectively. By contrast, an increase in FEV1 ≥ 12% and 200 mL demonstrated a sensitivity of 62.32%, specificity of 82.50%, and accuracy of 65.59% in diagnosing asthma. The changes of MEF25 and MEF50 before and after BD reversibility test may be of additional value in the clinical diagnosis of asthma, with cutoff values of 25% being the most.

Abnormal immunity of non-survivors with COVID-19: predictors for mortality.

BACKGROUND: The number of coronavirus disease 2019 (COVID-19) cases has rapidly increased all over the world. Specific information about immunity in non-survivors with COVID-19 is scarce. This study aimed to analyse the clinical characteristics and abnormal immunity of the confirmed COVID-19 non-survivors. METHODS: In this single-centered, retrospective, observational study, we enrolled 125 patients with COVID-19 who were died between January 13 and March 4, 2020 in Renmin Hospital of Wuhan University. A total of 414 randomly recruited patients with confirmed COVID-19 who were discharged from the same hospital during the same period served as control. The demographic, clinical characteristics and laboratory findings at admission, and treatment used in these patients were collected. The immunity-related risk factors associated with in-hospital death were tested by logistic regression models and Receiver Operating Characteristic (ROC) curve. RESULTS: Non-survivors (70 years, IQR: 61.5-80) were significantly older than survivors (54 years, IQR: 37-65) (P <  0.001). 56.8% of non-survivors was male. Nearly half of the patients (44.9%) had chronic medical illness. In non-survivors, hypertension (49.6%) was the most common comorbidity, followed by diabetes (20.0%) and coronary heart disease (16.0%). The common signs and symptoms at admission of non-survivors were fever (88%), followed by cough (64.8%), dyspnea (62.4%), fatigue (62.4%) and chest tightness (58.4%). Compared with survivors, non-survivors had higher white blood cell (WBC) count (7.85 vs 5.07 × 109/L), more elevated neutrophil count (6.41 vs 3.08 × 109/L), smaller lymphocyte count (0.69 vs 1.20 × 109/L) and lower platelet count (172 vs 211 × 109/L), raised concentrations of procalcitonin (0.21 vs 0.06 ng/mL) and CRP (70.5 vs 7.2 mg/L) (P < 0.001). This was accompanied with significantly decreased levels of CD3+ T cells (277 vs 814 cells/µl), CD4+ T cells (172 vs 473 cells/µl), CD8+ T cells (84 vs 262.5 cells/µl, P < 0.001), CD19+ T cells (88 vs 141 cells/µl) and CD16+ 56+ T cells (79 vs 128.5 cells/µl) (P < 0.001). The concentrations of immunoglobulins (Ig) G (13.30 vs 11.95 g/L), IgA (2.54 vs 2.21 g/L), and IgE (71.30 vs 42.25 IU/ml) were increased, whereas the levels of complement proteins (C)3 (0.89 vs 0.99 g/L) and C4 (0.22 vs 0.24 g/L) were decreased in non-survivors when compared with survivors (all P < 0.05). The non-survivors presented lower levels of oximetry saturation (90 vs 97%) at rest and lactate (2.40 vs 1.90 mmol/L) (P < 0.001). Old age, comorbidity of malignant tumor, neutrophilia, lymphocytopenia, low CD4+ T cells, decreased C3, and low oximetry saturation were the risk factors of death in patients with confirmed COVID-19. The frequency of CD4+ T cells positively correlated with the numbers of lymphocytes (r = 0.787) and the level of oximetry saturation (r = 0.295), Whereas CD4+ T cells were negatively correlated with age (r =-0.323) and the numbers of neutrophils (r = - 0.244) (all P < 0.001). CONCLUSIONS: Abnormal cellular immunity and humoral immunity were key features of non-survivors with COVID-19. Neutrophilia, lymphocytopenia, low CD4+ T cells, and decreased C3 were immunity-related risk factors predicting mortality of patients with COVID-19.

Application of Inflammatory Markers in Induced Sputum in Stable Chronic Obstructive Pulmonary Disease Patients with Positive Bronchodilation Tests.

Positive bronchodilation (BD) tests can be noticed in some stable chronic obstructive pulmonary disease (COPD) patients. The characteristics of airway inflammation in this entity remain unclear. Our study aimed to identify the characteristics of airway inflammation in stable COPD patients with positive BD tests. The airway inflammation was assessed in 88 patients with stable COPD using the examination of induced sputum in the aftermath of lung function and BD tests. Cellular counts and the levels of molecular markers including eosinophil cationic protein (ECP), myeloperoxidase (MPO), interleukin-5 (IL-5), and IL-8 were assayed by Wright's stain, Immuno-CAP system, and ELISA, RT-PCR. Among the 88 patients with stable COPD, 20 (22.7%) showed positive BD tests. The values of eosinophils (4.7%±3.4%) and ECP (90.1±41.6 ng/mL) in induced sputum in stable COPD patients with positive BD tests were markedly elevated as compared with those in stable COPD patients with negative BD tests or in healthy controls (all P>0.05), but significantly lower than those in asthmatic patients (all P<0.01). The IL-5 in sputum supernatant was significantly decreased in stable COPD patients with positive BD tests as compared with the patients with asthma (12.5±7.8 vs. 48.2±26.0 ng/mL;.P<0.01). However, healthy controls exhibited similar concentrations of IL-5 in induced sputum with patients with stable COPD, whether with positive or negative BD tests (all P>0.05). Moreover, the values of neutrophils (61.8%±15.1%), MPO (574.0±111.8 ng/mL), and IL-8 (32.6±13.4 ng/mL) in induced sputum in stable COPD patients with positive BD tests were significantly higher than those in asthmatics or normal controls (all P<0.01). However, the values of the above inflammatory markers in induced sputum were similar among stable COPD patients with positive or negative BD tests (all P>0.05). The stable COPD patients with positive BD tests may present not only eosinophilic airway inflammation but also neutrophilic airway inflammation.

[Sulfatide-loaded CD1d tetramer to detect typeII NKT cells in mice].

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's disease misdiagnosed as pneumonia: two case reports.

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.

Increased expression of CD86 and reduced production of IL-12 and IL-10 by monocyte-derived dendritic cells from allergic asthmatics and their effects on Th1- and Th2-type cytokine balance.

BACKGROUND: In allergic asthma, allergen-specific T cells have a Th2-biased phenotype, and it is thought that dendritic cells (DCs) contribute to the induction of allergic immune responses. Therefore, we hypothesized that DCs from allergic asthmatics and healthy donors differ with regard to their preference to induce Th1 or Th2 immune responses. OBJECTIVES: To investigate differences in DC-expressed costimulatory molecules and DC-secreted cytokines between allergic asthmatics and healthy donors, and their influence on the Th1- and Th2-type cytokine balance. METHODS: Circulating monocytes from patients with allergic asthma and healthy donors were cultured with GM-CSF and IL-4, respectively, for 5 days and subsequently with lipopolysaccharide for 2 days to create mature DCs (mDCs). CD1a, CD83, CD40 and CD86 expression on mDCs was examined using a fluorescence-activated cell sorter. IL-12 and IL-10 secreted by mDCs were measured by ELISA. Naïve cord blood T cells were primed by mDCs from two groups, and IL-4 and IFN-gamma production by polarized T-helper cells (Th) was measured by ELISA. RESULTS: (1) CD86 expression on mDCs from allergic asthmatics was higher than that from healthy donors. (2) IL-12, IL-12p40 and IL-10 production by mDCs from allergic asthmatics was significantly lower than that from healthy donors, respectively. (3) IL-4 production by Th cells primed by mDCs from allergic asthmatics was increased compared with that from healthy donors. CONCLUSIONS: mDCs from allergic asthmatics preferentially priming naïve T cells towards Th2-cell development might be due to increased expression of CD86 and reduced production of IL-12 and IL-10.

[A study of phenotype and function of dendritic cells and secretory cytokine in allergic asthmatic patients].

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.

Interleukin-12 was not involved in promotion of T helper cell differentiation induced by theophylline.

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.

Effects of theophylline on CD4+ T lymphocyte, interleukin-5, and interferon gamma in induced sputum of asthmatic subjects.

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.