Selective production of IL-33-neutralizing autoantibody ameliorates asthma responses and severity.

BACKGROUND: Natural anti-cytokine autoantibodies can regulate homeostasis of infectious and inflammatory diseases. The anti-cytokine autoantibody profile and relevance to the pathogenesis of asthma are unknown. We aim to identify key anti-cytokine autoantibodies in asthma patients, and reveal their immunological function and clinical significance. METHODS: A Luciferase Immunoprecipitation System was used to screen serum autoantibodies against 11 key cytokines in patients with allergic asthma and healthy donors. The antigen-specificity, immunomodulatory functions and clinical significance of anti-cytokine autoantibodies were determined by ELISA, qPCR, neutralization assays and statistical analysis, respectively. Potential conditions for autoantibody induction were revealed by in vitro immunization. RESULTS: Of 11 cytokines tested, only anti-IL-33 autoantibody was significantly increased in asthma, compare to healthy controls, and the proportion positive was higher in patients with mild-to-moderate than severe allergic asthma. In allergic asthma patients, the anti-IL-33 autoantibody level correlated negatively with serum concentration of pathogenic cytokines (e.g., IL-4, IL-13, IL-25 and IL-33), IgE, and blood eosinophil count, but positively with mid-expiratory flow FEF25-75%. The autoantibodies were predominantly IgG isotype, polyclonal and could neutralize IL-33-induced pathogenic responses in vitro and in vivo. The induction of the anti-IL-33 autoantibody in blood B-cells in vitro required peptide IL-33 antigen along with a stimulation cocktail of TLR9 agonist and cytokines IL-2, IL-4 or IL-21. CONCLUSIONS: Serum natural anti-IL-33 autoantibodies are selectively induced in some asthma patients. They ameliorate key asthma inflammatory responses, and may improve lung function of allergic asthma.

qSOFA predicted pneumonia mortality better than minor criteria and worse than CURB-65 with robust elements and higher convergence.

BACKGROUND: Limited data are available on the discriminatory capacity of quick sequential [sepsis-related] organ failure assessment (qSOFA) versus IDSA/ATS minor criteria for predicting mortality in patients with community-acquired pneumonia (CAP). METHODS: An observational prospective cohort study of 2116 patients with CAP was performed. Construct validity was determined using Cronbach α. Discrimination was assessed using the area under the receiver operating characteristic curve (AUROC) and net reclassification improvement (NRI). RESULTS: Overall in-hospital mortality was 6.43%. Mortality was 25.96% for patients with a qSOFA score of 2 or higher versus 3.05% for those with a qSOFA score less than 2 (odds ratio for mortality 6.57, P < 0.0001), and 13.85% for patients with at least 3 minor criteria versus 2.03% for those with 2 or fewer minor criteria (odds ratio for mortality 2.27, P < 0.0001). qSOFA had a higher correlation with mortality than minor criteria, as well as higher internal consistency (Cronbach alpha 0.43 versus 0.14) and diagnostic values of individual elements (larger AUROCs and higher Youden's indices). qSOFA ≥2 was less sensitive but more specific for predicting mortality than ≥3 minor criteria (qSOFA sensitivity 59.6%, specificity 88.3% and positive likelihood ratio 5.11 versus ≥3 minor criteria sensitivity 80.1%, specificity 65.8% and positive likelihood ratio 2.34). The predictive validity of qSOFA was good for mortality (AUROC = 0.868), was statistically greater than minor criteria, was equal to pneumonia severity index, and was inferior compared with CURB-65 (AUROC, 0.824, 0.902, 0.919; NRI, 0.088, -0.068, -0.103; respectively). CONCLUSIONS: The qSOFA predicted mortality in CAP better than IDSA/ATS minor criteria and worse than CURB-65 with robust elements and higher convergence. qSOFA as a bedside prompt might be positioned as a proxy for minor criteria and increase the recognition and thus merit more appropriate management of CAP patients likely to fare poorly, which might have implications for more accurate clinical triage decisions.

Cold-inducible RNA-binding protein might determine the severity and the presences of major/minor criteria for severe community-acquired pneumonia and best predicted mortality.

BACKGROUND: Severity of community-acquired pneumonia (CAP) depends on microbial pathogenicity, load and virulence, and immune responses. The Infectious Disease Society of America and the American Thoracic Society (IDSA/ATS) minor criteria responsible for clinical triage of patients with CAP are of unequal weight in predicting mortality. It is unclear whether the IDSA/ATS major/minor criteria might be strongly and positively associated with the immune responses. It is warranted to explore this intriguing hypothesis. METHODS: A prospective cohort study of 404 CAP patients was performed. Cold-inducible RNA-binding protein (CIRP) levels were measured using a sandwich-based enzyme-linked immunosorbent assay. The receiver operating characteristic curves were created and the areas under the curves were calculated to illustrate and compare the accuracy of the indices. RESULTS: Severe CAP patients meeting the major criteria had the highest plasma concentrations of CIRP. The more the number of most predictive minor criteria strongly associated to mortality, i.e. arterial oxygen pressure/fraction inspired oxygen ≤ 250 mmHg, confusion, and uremia, present, the higher the CIRP level. Interestingly, the patients with non-severe CAP meeting the most predictive minor criteria demonstrated unexpectedly higher CIRP level compared with the patients with severe CAP not fulfilling the criteria. Procalcitonin (PCT), interleukin-6 (IL-6), C-reactive protein (CRP), sequential organ failure assessment (SOFA) and pneumonia severity index (PSI) scores, and mortality confirmed similar intriguing patterns. CIRP was strongly linked to PCT, IL-6, CRP, minor criteria, SOFA and PSI scores, and mortality (increased odds ratio 3.433). The pattern of sensitivity, specificity, positive predictive value, and Youden's index of CIRP ≥ 3.50 ng/mL for predicting mortality was the optimal. The area under the receiver operating characteristic curve of CIRP was the highest among the indices. CONCLUSIONS: CIRP levels were strongly correlated with the IDSA/ATS major/minor criteria. CIRP might determine the severity and the presences of major/minor criteria and best predicted mortality, and a CIRP of ≥ 3.50 ng/mL might be more valuable cut-off value for severe CAP, suggesting that CIRP might be a novel and intriguing biomarker for pneumonia to monitor host response and predict mortality, which might have implications for more accurate clinical triage decisions.

Correction to: Scored minor criteria for severe community-acquired pneumonia predicted better.

.

Scored minor criteria for severe community-acquired pneumonia predicted better.

BACKGROUND: Infectious Disease Society of America/American Thoracic Society (IDSA/ATS) minor criteria for severe community-acquired pneumonia (CAP) are of unequal weight in predicting mortality, but the major problem associated with IDSA/ATS minor criteria might be a lack of consideration of weight in prediction in clinical practice. Would awarding different points to the presences of the minor criteria improve the accuracy of the scoring system? It is warranted to explore this intriguing hypothesis. METHODS: A total of 1230 CAP patients were recruited to a retrospective cohort study. This was tested against a prospective two-center cohort of 1749 adults with CAP. 2 points were assigned for the presence of PaO2/FiO2 ≤ 250 mmHg, confusion, or uremia on admission and 1 point for each of the others. RESULTS: The mortality rates, and sequential organ failure assessment (SOFA) and pneumonia severity index (PSI) scores increased significantly with the numbers of IDSA/ATS minor criteria present and minor criteria scores. The correlations of the minor criteria scores with the mortality rates were higher than those of the numbers of IDSA/ATS minor criteria present. As were the correlations of the minor criteria scores with SOFA and PSI scores, compared with the numbers of IDSA/ATS minor criteria present. The pattern of sensitivity, specificity, positive predictive value, and Youden's index of scored minor criteria of ≥2 scores or the presence of 2 or more IDSA/ATS minor criteria for prediction of mortality was the best in the retrospective cohort, and the former was better than the latter. The validation cohort confirmed a similar pattern. The area under the receiver operating characteristic curve of scored minor criteria was higher than that of IDSA/ATS minor criteria in the retrospective cohort, implying higher accuracy of scored version for predicting mortality. The validation cohort confirmed a similar paradigm. CONCLUSIONS: Scored minor criteria orchestrated improvements in predicting mortality and severity in patients with CAP, and scored minor criteria of ≥2 scores or the presence of 2 or more IDSA/ATS minor criteria might be more valuable cut-off value for severe CAP, which might have implications for more accurate clinical triage decisions.

Benzo(a)pyrene facilitates dermatophagoides group 1 (Der f 1)-induced epithelial cytokine release through aryl hydrocarbon receptor in asthma.

BACKGROUND: Environmental pollutants, which coexist with allergens, have been associated with the exacerbation of asthma. However, the underlying molecular mechanisms remain elusive. We sought to determine whether benzo(a)pyrene (BaP) co-exposure with dermatophagoides group 1 allergen (Der f 1) can potentiate Der f 1-induced asthma and its underlying mechanisms. METHODS: The effect of BaP was investigated in Der f 1-induced mouse model of asthma, including airway hyper-responsiveness, allergic inflammation, and epithelial-derived cytokines. The impact of BaP on Der f 1-induced airway epithelial cell oxidative stress (ROS) and cytokine release was further analyzed. The role of aryl hydrocarbon receptor (AhR) signaling in BaP-promoted Der f 1-induced ROS, cytokine production, and allergic inflammation was also investigated. RESULTS: Compared with Der f 1, BaP co-exposure with Der f 1 led to airway hyper-responsiveness and increased lung inflammation in mouse model of asthma. Increased expression of TSLP, IL-33, and IL-25 was also found in the airways of these mice. Moreover, BaP co-exposure with Der f 1 activated AhR signaling with increased expression of AhR and CYP1A1 and promoted airway epithelial ROS generation and TSLP and IL-33, but not IL-25, expression. Interestingly, AhR antagonist CH223191 or cells with AhR knockdown abrogated the increased expression of ROS, TSLP, and IL-33. Furthermore, ROS inhibitor N-acetyl-L-cysteine (NAC) also suppressed BaP co-exposure-induced expression of epithelial TSLP, IL-33, and IL-25. Finally, AhR antagonist CH223191 and NAC inhibited BaP co-exposure with Der f 1-induced lung inflammation. CONCLUSIONS: Our findings suggest that BaP facilitates Der f 1-induced epithelial cytokine release through the AhR-ROS axis.

Rapid and specific detection of porcine parvovirus using real-time PCR and high resolution melting (HRM) analysis.

BACKGROUND: Porcine parvovirus (PPV) is the important causative agent for infectious infertility, which is a fairly tough virus that multiplies normally in the intestine of pigs without causing clinical signs in the world. RESULTS: We developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the detection of PPV. Primers targeting the VP gene were highly specific, as evidenced by the negative amplification of closely related viruses, such as porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The performance of unlabeled real time PCR was compared to TaqMan real time PCR, and the detection limits of the two methods were nearly equal. Moreover, there was good correlation between Cp and diluted genomic DNA when tested with the two methods. The assay has the accuracy of 100% in reference to labeled real time PCR, when it was tested on 45 clinical samples. CONCLUSIONS: The present study demonstrated that the established assay integrating real-time PCR and HRM is relatively cost-effective and more stable, which provides an alternative tool for rapid, simple, specific and sensitive detection of PPV.

Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini using real-time PCR and high resolution melting analysis.

Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.

Endoplasmic reticulum stress drives macrophages to produce IL-33 to favor Th2 polarization in the airways.

Interleukin (IL)-33 is a key driver of T helper 2 (Th2) cell polarization. Endoplasmic reticulum (ER) stress plays a role in the skewed T cell activation. The objective of this project is to elucidate the role of IL-33 derived from macrophages in inducing Th2 polarization in the airways. In this study, bronchoalveolar lavage fluids (BALF) were collected from patients with asthma and healthy control subjects. Macrophages were isolated from the BALF by flow cytometry cell sorting. An asthmatic mouse model was established using the ovalbumin/alum protocol. The results showed that increased IL33 gene activity and ER stress-related molecules in BALF-derived M2a macrophages was observed in asthmatic patients. Levels of IL33 gene activity in M2a cells were positively correlated with levels of asthma response in asthma patients. Sensitization exacerbated the ER stress in the airway macrophages, which increased the expression of IL-33 in macrophages of airway in sensitized mice. Conditional ablation of Il33 or Perk or Atf4 genes in macrophages prevented induction of airway allergy in mice. In conclusion, asthma airway macrophages express high levels of IL-33 and at high ER stress status. Inhibition of IL-33 or ER stress in macrophages can effectively alleviate experimental asthma.

ER stress modulates the immune regulatory ability in gut M2 cells of patients with ulcerative colitis.

This study aims to characterize the impaired immune regulatory function of Mφ obtained from UC patient colon lavage fluid (CLF). Mφs were the largest proportion (21.3 4.0%) of the CLF-derived cellular components. Less abundant and weaker immune suppressive function were observed in M2 Mφs (M2 cells) of the ulcerative colitis (UC) group. High levels of endoplasmic reticulum (ER) stress associated molecules were detected in UC M2 cells. The spliced X box binding protein-1 (XBP1) gene was negatively correlated with programmed death ligand-1 (PD-L1) in UC M2 cells. XBP1 promoted the expression of ring-finger protein 20 (Rnf20) in M2 cells. Rnf20 reduced PD-L1 abundance in UC M2 cells and impaired the immune suppressive ability. Inhibition of Rnf20 restored the immune regulating capacity of M2 cells and suppressed experimental colitis.

Single cell characteristics of patients with vaccine-related adverse reactions following inactivated COVID-19 vaccination.

A good safety and immunogenicity profile was reported in Phase I and II clinical trials of inactivated SARS-CoV-2 vaccines. Here, we report two cases associated with vaccine-associated adverse events, including one patient with fever and another with anaphylactic shock resulting from inactivated SARS-CoV-2 vaccination. Cell sub-types and the importance of genetic characteristics were assessed using single-cell mRNA sequencing and machine learning. Overall, the patient with fever showed a significant increase in the numbers of cytotoxic CD8 T cells and MKI67high CD8 T cells. A potential concurrent infection with the Epstein-Barr virus enhanced interferon type I responses to vaccination against the virus. STAT1, E2F1, YBX1, and E2F7 played a key role in the transcription regulation of MKI67high CD8 T cells. In contrast, the patient with allergic shock displayed predominant increases in the numbers of S100A9high monocytes, activated CD4 T cells, and PPBPhigh megakaryocytes. The decision tree showed that LYZ and S100A8 in S100A9high monocytes contributed to the degranulation of neutrophils and activation of neutrophils involved in allergic shock. PPBP and PF4 were major contributors to platelet degranulation. These findings highlight the diversity of adverse reactions following inactivated SARS-CoV-2 vaccination and show the emerging role of cellular subtypes and central genes in vaccine-associated adverse reactions.

The identification of cell sub-types may help in the diagnosis of COVID-19 vaccine-related adverse events.COVID-19 vaccination-related acute pulmonary edema may induce a higher risk of thrombosis.The long-term fever after vaccination may attribute to the excessive type I interferon responses.

Updating cut-off values of severity scoring systems for community-acquired pneumonia to orchestrate more predictive accuracy.

Background: The assessment of severity is crucial in the management of community-acquired pneumonia (CAP). It remains unknown whether updating cut-off values of severity scoring systems orchestrate improvement in predictive accuracy.Methods: 3,212 patients with CAP were recruited to two observational prospective cohort studies. Three bettered scoring systems were derived from the corresponding well-established and extensively used pneumonia-specific severity scoring systems, i.e. pneumonia severity index, minor criteria and CURB-65 (confusion, urea >7 mmol/L, respiratory rate ≥30/min, low blood pressure, and age ≥65 years) score, with the updating cut-off values for tachypnea and low blood pressure. Cronbach α was employed to determine construct validity. Discrimination was valued by calculating the area under the receiver operating characteristic curve (AUROC) and net reclassification improvement (NRI).Results: Respiratory rate ≥22/min and systolic blood pressure ≤100 mm Hg were performed better than respiratory rate ≥30/min and hypotension for predicting mortality in the derivation cohort, respectively (AUROC, 0.823 vs 0.519, 0.688 vs 0.622; NRI, 0.61, 0.13). Bettered scoring systems orchestrated higher convergences, indicated by greater Cronbach α and more decrease in Cronbach α if the updating cut-off values were deleted. The six scoring systems agreed well with one another. Bettered- pneumonia severity index, minor criteria and CURB-65 score showed higher associations with severity and mortality rates and demonstrated greater predictive accuracies for mortality compared with the corresponding original systems (AUROC, 0.939 vs 0.883, 0.909 vs 0.871, 0.913 vs 0.859; NRI, 0.113, 0.076, 0.108; respectively). The validation cohort confirmed a similar pattern.Conclusions: Updating cut-off values of severity scoring systems for CAP orchestrate improvement in predictive accuracy, suggesting that it may facilitate the rationalization of clinical triage decision-making and further reduce mortality. The current studies provide the first known prospective evidence of potential benefit of the updating cut-off values of severity scoring systems for CAP in predictive accuracy.Key messagesUpdating cut-off values were performed better for predicting mortality.Bettered scoring systems orchestrated higher convergences.Bettered scoring systems demonstrated greater predictive accuracies for mortality.

[Study of tacrolimus intranasal treatment for allergic asthma in mice].

OBJECTIVE: To explore the efficacy of intranasal treatment by immunosuppressant tacrolimus for allergic asthma and its mechanism in mice. METHODS: 24 female BALB/c mice were randomly divided into 4 groups: group A (negative control), group B (model control), group C (low dose treatment), and group D (high dose treatment). Mice in group A were treated with saline (100 microl). Other groups were sensitized intraperitoneally with allergen extracts of Dermatophagoides farinae (Der f) absorbed to Al(OH)3 at day 0, 7, and 14. From day 28, groups A, B, C, and D were intranasally treated with saline, PBS, 0.01% tacrolimus, and 0.1% tacrolimus, respectively, once per day for 7 d, and followed by intranasal challenge with 50 microl Der f extracts in the mean time. 24 h after the last challenge, the airway hyper-responsiveness (AHR) were detected. At 48 h after the last challenge, the mice were sacrificed, the bronchoalveolar lavage fluid (BALF) was collected, the lungs and spleen were aseptically removed. The total cell number and cell classification of BALF were recorded. The level of interleukin-4 (IL-4), interleukin-5 (IL-5), interferon-gamma (IFN-gamma) in BALF and in spleen cells culture supernatants was detected by ELISA. The lung inflammation and mucus secretion were observed in mice by HE (haematoxylin and eosin) staining and AB (Alcian Blue) staining. RESULTS: Compared with group B, AHR (P < 0.05) and airway inflammation in group D significantly reduced. The number of total cells [(29.92 +/- 5.20) x 10(4)/ml] (P < 0.05) and eosinophils [(4.33 +/- 0.75) x 10(4)/ml] (P < 0.01) in group D greatly decreased than those of group B [(59.33 +/- 5.99) x 10(4)/ml and (22.67 +/- 5.65) x 10(4)/ml]. The level of IL-4 [(22.49 +/- 4.96) pg/ml] (P < 0.05), IL-5 [(43.90 +/- 13.15) pg/ml] (P < 0.01) and IFN-gamma [(10.17 +/- 1.09) pg/ml] (P < 0.05) in BALF significantly decreased (P < 0.05) than those of group B [(57.02 +/- 7.38), (133.49 +/- 15.63) and (15.32 +/- 3.23) pg/ml, respectively]. The level of IL-4 [(22.54 +/- 4.58) pg/ml], IL-5 [(3631 +/- 20.85) pg/ml] and IFN-gamma [(11.28 +/- 1.79) pg/ml] in spleen cell culture supernatant all significantly decreased (P < 0.05) than those of group B [(56.34 +/- 6.21), (72.3 +/- 6.23) and (18.82 +/- 1.88) pg/ml, respectively]. There was no significant difference between group C and group B. CONCLUSION: Tacrolimus shows certain immune therapeutic effect on dust mite sensitized mice, and this effect may be attributed to its inhibition on T lymphocyte factor secretion.

[Therapeutic effect on murine asthma with sublingual use of Dermatophagoides farinae/chitosan nanoparticle vaccine].

OBJECTIVE: To prepare Dermatophagoides farinae (Der f)/chitosan nanoparticle vaccine (DCN), and to investigate the effect of sublingual administration with DCN in asthma mice model. METHODS: DCN were prepared by ionotropic gelation. 30 BALB/c mice were randomly divided into 5 groups: normal control group(A), PBS control group (B), Chitosan group (C), Der f group (D), DCN group (E). Group A were treated with normal saline (100 microl) all the time. Mice in other groups were sensitized intraperitoneally with 50 microg dust mite extracts plus 2 mg Al(OH)3, and on day 28 given a sublingual vaccination of PBS(group B), or empty CS nanoparticles (group C), or Der f (group D, 1 mg Der f) or DCN (group E, loaded with 1 mg Der f). All the mice received 18 doses at 1-day intervals. One week after the last immunization, mice in group B, C, D, and E were intranasally challenged with 50 microg Der f extract daily for seven days. Twenty-four hours after the last challenge, airway hyper-responsiveness (AHR) was assessed by using whole-body plethysmography. Two days post challenge, mice were sacrificed and bronchoalveolar lavage fluid (BALF) was collected. Number of the total cells and eosinophils was determined. Level of cytokines in the supernatant of splenocyte culture was assayed by ELISA. Level of Der f specific IgE, IgG2a and IgA in the sera was determined by ELISA. Airway inflammation was analyzed by HE staining. Spleen lymphocyte proliferation responses were analyzed by MTT colorimetry. RESULTS: Compared with group B, AHR and the lung inflammation in groups D and E were greatly reduced. Numbers of total cells and eosinophils in BALF of groups D (36.50 x 10(4)/ml, 3.72 x 10(4)/ml) and E (34.25 x 10(4)/ml, 2.25 x 10(4)/ml) were significantly lower than that of group B (61.67 x 10(4)/ml, 14.17 x 10(4)/ml) (P < 0.05). The level of specific IgE was significantly lower in groups D (0.22) and E (0.22), and that of IgA in groups D (0.88) and E (1.03) was significantly higher than that in group B (0.79). The level of IL-4 in BALF (D: 28.49 pg/ml, E: 20.93 pg/ml) and cultured splenocytes (D: 27.82 pg/ml, E: 20.80 pg/ml) of groups D and E was significantly lower than that of group B (56.33 pg/ml, 45.84 pg/ml) (P < 0.05). While IFN-gamma (D: 18.80 pg/ml, E: 37.32 pg/nml) and IL-10 (D: 118.90 pg/ml, E: 129.15 pg/ml) in BALF in groups D and E were significantly higher than that of group B (13.60 pg/ml, 29.61 pg/ml) (P < 0.05), and same with IFN-y (D: 20.68 pg/ml, E: 42.42 pg/ml) and IL-10 (D: 36.31 pg/ml, E: 161.37 pg/ml) in spleen cultured supernatants of groups D and E (P < 0.05). The allergen-specific splenocyte proliferation was inhibited in groups D (SI: 0.14) and E (SI: 0.13), and there was no significant difference between group C (SI: 0.22) and group B (SI: 0.23). CONCLUSION: Dermatophagoides farinae (Der f)/chitosan nanoparticle vaccine has therapeutic effect on murine asthma.

Sublingual immunotherapy efficacy of Dermatophagoides farinae vaccine in a murine asthma model.

BACKGROUND: Allergen-specific sublingual immunotherapy is a potential treatment for allergic diseases. Its effective dose and underlying mechanism are still to be explored. Here, we investigated the efficacy and mechanism of sublingually administered Dermatophagoides farinae (Der f) vaccine in a murine asthma model. METHODS: BALB/c mice were sensitized intraperitoneally with Der f extract absorbed to alum, followed by sublingual treatment with Der f vaccine for 6 weeks. The mice were subsequently challenged intranasally with Der f extract for 1 week. We analyzed their clinical symptoms, antibody levels, cytokine levels, T-cell proliferation and the regulatory T-cell numbers. RESULTS: Mice treated with high-dose Der f sublingual vaccine prior to challenge displayed alleviated symptoms such as airway hyperreactivity, lung inflammation and mucus production, as well as less eosinophilic cells in bronchoalveolar lavage fluid. Interestingly, reduced responses of Der-f-specific IgE and increased responses of Der-f-specific IgA and IgG1 were aroused in the high-dose Der f sublingual vaccine group. We also observed that interleukin-4 was reduced and interferon-gamma and interleukin-10 were increased among splenocytes and in bronchoalveolar lavage fluid, which inhibited Der-f-specific T-cell proliferation of the spleen and increased CD4+CD25+Foxp3+regulatory T cells in the spleen. However, mice treated with low-dose Der f sublingual vaccine developed allergic asthma. CONCLUSION: Our results illustrate that high-dose Der f sublingual vaccine may play a role in immunologic protection in murine allergic asthma, possibly by inducing regulatory T cells and Th1 reaction.

Feasibility analysis of flexible bronchoscopy in conjunction with noninvasive ventilation for therapy of hypoxemic patients with Central Airway Obstruction: a retrospective study.

BACKGROUND: Interventional bronchoscopy for hypoxemic patients with central airway obstruction (CAO) is typically performed under general anesthesia. This approach poses remarkable challenge for both bronchoscopist and anesthesiologist. Noninvasive ventilation (NIV) during flexible bronchoscopy (FB) has been successfully used in hypoxemic patients, but rarely in the treatment of hypoxemic patients with CAO. OBJECTIVE: To evaluate the feasibility of therapeutic FB assisted with NIV for therapy of hypoxemic patients with CAO. METHOD: Twenty-nine hypoxemic CAO patients treated with FB from December 2010 to May 2016 in our hospital were retrospectively reviewed, either aided with NIV under sedation (NIV group ) or through artificial airway under general anesthesia (control group). Interventional procedures included balloon dilation, electrocautery and argon plasma coagulation. RESULT: Fifteen patients were enrolled in the NIV group and 14 in the control group. The success rate (93.3% VS 92.9%, p = 1.0), procedure time (60.5 ± 4.2 min VS 67.8 ± 5.6 min, p = 0.31) and oxygenation improvement between the two groups have no significant difference. Less reduction of systolic blood pressure and heart rate during procedure was observed in the NIV group. The NIV group showed shorter admission time before procedure than the control group (35.1 ± 4.6 h VS 55.6 ± 5.6 h, p < 0.01). In addition, procedure fee in the NIV group was significantly less than that in the control group (540.7 ± 62.8$ VS975.4 ± 69.5$, p < 0.0001). CONCLUSION: FB assisted with NIV is a safe, efficient and economic method for therapy of selected hypoxemic patients with CAO.

Survivin Impairs the Apoptotic Machinery in CD4+ T Cells of Patients with Ulcerative Colitis.

BACKGROUND: The increase in CD4+ T cell infiltration and overproduction of CD4+ T cell-associated cytokines have been observed in the inflamed colon mucosa of patients with ulcerative colitis (UC); the underlying mechanisms are not fully understood. Survivin plays a critical role in the interference with apoptotic machinery. This study aims to elucidate the role of survivin in the interference with the apoptotic machinery in CD4+ T cells of UC patients. METHODS: Peripheral blood samples were collected from UC patients (UC group) and healthy subjects (healthy group). The apoptotic status in CD4+ T cells was analyzed by flow cytometry. RESULTS: We observed that the expression of survivin was significantly higher in CD4+ T cells of UC patients than in healthy subjects. UC CD4+ T cells were resistant to apoptosis induction. A complex of survivin and c-Myc, the transcription factor of FasL, was detected in CD4+ T cells in UC patients, which prevented the binding of c-Myc to the FasL promoter and interfered with the expression of FasL. Increased expression of survivin prevented the activation-induced CD4+ T cells from apoptosis. CONCLUSIONS: The data indicate that UC CD4+ T cells express high levels of survivin, which impairs the apoptotic machinery in CD4+ T cells and prevents the activation-induced CD4+ T cell apoptosis. Therefore, target therapy against survivin has translational potential in the treatment of UC patients.

Local nasal immunotherapy: efficacy of Dermatophagoides farinae-chitosan vaccine in murine asthma.

BACKGROUND: Local nasal immunotherapy has been reported to be effective for airway allergic diseases. A biodegradable material, chitosan (CS), has been reported to be safe and effective in allergen delivery. In this study, we tested immunotherapeutic efficiency by intranasal administration of Der f entrapped in CS microparticles to sensitized mice. METHODS: BALB/c mice were sensitized intraperitoneally with Der f extract absorbed to alum, followed by intranasal treatment with PBS, CS, Der f or Der f-CS nano-vaccine for 6 weeks. The mice were subsequently challenged intranasally with Der f extract for 1 week, and we analyzed their clinical symptoms, antibody expression levels, cytokine levels, T cell proliferation and regulatory T cell numbers. RESULTS: Mice treated with intranasal Der f-CS nano-vaccine prior to challenge displayed an alleviated spectrum of symptoms including airway hyper-reactivity, lung inflammation and mucus production and had fewer eosinophilic cells in bronchoalveolar lavage fluid (BALF). Interestingly, the cytokine levels in Der f-specific IgE were reduced, but IgA in serum and BALF was increased. We also observed that IL-4 was reduced and IFN-gamma and IL-10 were increased among splenocytes and in BALF, which inhibits Der f-specific T-cell proliferation in splenocytes and increases regulatory T cells in the spleen. However, the mice challenged without intranasal Der f or Der f-CS vaccine treatment developed allergic asthma. CONCLUSION: Our results illustrate that intranasal administration of Der f-CS nano-vaccine plays roles in immunologic protection in murine allergic asthma by inducing regulatory T cells and Th1-type reaction.