Yves Pacheco

Dendritic Cells Infiltrating Human Non-Small Cell Lung Cancer Are Blocked at Immature Stage

The efficacy of immune response to control human cancer remains controversial. It is particularly debated whether and to what extent the capacity of tumor-infiltrating dendritic cells (DC) to drive immunization can be turned off by transformed cells, leading to tumor-specific tolerance rather than immunization. To address this issue, we have characterized the DC isolated from human non-small cell lung cancer (NSCLC). These biopsy specimens contained CD11chigh myeloid DC (mDC), but also CD11c− plasmacytoid DC (pDC) and a third DC subset expressing intermediate level of CD11c. Compared with peripheral blood, CD11chigh tumor-infiltrating DC (TIDC) displayed a “semi-mature” phenotype, and TLR4 or TLR8 stimulation drove them to mature partially and to secrete limited amounts of cytokines. In contrast, most tumor-infiltrating pDC were immature but underwent partial maturation after TLR7 activation, whereas TLR9 ligation triggered low secretion of IFN-α. CD11cint mDC represented ∼25% of total DC in tumoral and peritumoral tissues and expressed low levels of costimulatory molecules contrasting with high levels of the immunoinhibitory molecule B7-H1. Finally, the poor APC function of total TIDC even after TLR stimulation and the migratory response of both tumor-infiltrating mDC and pDC toward CCL21 and SDF-1 in vitro suggested their ability to compromise the tumor-specific immune response in draining lymph nodes in vivo. Further studies will be required to establish the specific role of the three TIDC subsets in tumor immunity and to draw conclusions for the design of therapeutic strategies.

Elevated MMP-12 protein levels in induced sputum from patients with COPD

Background: Several matrix metalloproteinases (MMPs) are involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). In mice, MMP-12 plays a crucial role in the development of cigarette smoke induced emphysema. A study was undertaken to investigate the role of MMP-12 in the development of COPD in human smokers. Methods: Induced sputum samples were collected from patients with stable COPD (n = 28), healthy smokers (n = 14), never smokers (n = 20), and former smokers (n = 14). MMP-12 protein levels in induced sputum were determined by ELISA and compared between the four study groups. MMP-12 enzymatic activity in induced sputum was evaluated by casein zymography and by cleaving of a fluorescence quenched substrate. Results: Median (IQR) MMP-12 levels were significantly higher in COPD patients than in healthy smokers, never smokers, and former smokers (17.5 (7.1–42.1) v 6.7 (3.9–10.4) v 4.2 (2.4–11.3) v 6.1 (4.5–7.6) ng/ml, p = 0.0002). MMP-12 enzymatic activity was significantly higher in patients with COPD than in controls (4.11 (1.4–8.0) v 0.14 (0.1–0.2) μg/μl, p = 0.0002). Conclusion: MMP-12 is markedly increased in induced sputum from patients with stable COPD compared with controls, suggesting a role for MMP-12 in the development of COPD in smokers.

Expression of Class III β-Tubulin Is Predictive of Patient Outcome in Patients with Non–Small Cell Lung Cancer Receiving Vinorelbine-Based Chemotherapy

Purpose: To determine the prevalence and the prognostic value of microtubule component expression in tumors of patients with locally advanced or metastatic non–small cell lung cancer (NSCLC). Experimental Design: Expression of microtubular components was immunohistochemically examined in 93 tumor samples from untreated patients with stage III and IV NSCLC. All patients received vinorelbine-based chemotherapy. Response to chemotherapy, progression-free survival, and overall survival were correlated with the expression of microtubule proteins. Results: The response rate was 27.3% (21 partial responses among 77 valuable patients). Although expression of microtubule components was not associated with the response rate, high class III β-tubulin expression was correlated with resistance to vinorelbine, defined as disease progression under treatment. Patients whose tumors expressed high levels of class III β-tubulin isotype had shorter progression-free survival and overall survival (P = 0.002 and 0.001, respectively). High Δ2 α-tubulin expression was associated with a shorter overall survival (P = 0.018). Tubulin II levels were not found to be correlated with patient outcome. A multivariate analysis, taking into account sex, age, histology, stage, weight loss, and class II β-tubulin, class III β-tubulin, and Δ2 α-tubulin levels, confirmed that class III β-tubulin expression was independently correlated with progression-free survival (P = 0.04) and overall survival (P = 0.012). Conclusions: These findings suggest that a high level of expression of class III β-tubulin in tumor cells is associated with resistance to vinorelbine and a poor prognosis in patients with NSCLC receiving vinorelbine-based chemotherapy.

Quality of life during pollen season in patients with seasonal allergic rhinitis with or without asthma

Objectives: We studied the evolution of generic and rhinoconjunctivitis-specific quality of life (QOL) during pollen season in patients with isolated seasonal allergic rhinitis (SAR) and those with asthma and concomitant SAR (AS+SAR). Generic QOL between groups was also compared at pollen peak. Methods: A prospective cohort study was conducted in Southern France in 2002. Outpatients aged 18–60, regularly visiting respiratory physicians for SAR, were recruited before the grass (grass cohort) or ragweed pollination period (ragweed cohort). Before the pollination period (baseline) and at peak pollination, patients completed French versions of the Mini Rhinoconjuctivitis Quality of Life Questionnaire (Mini-RQLQ) and physical and mental Short Form-12 (SF-12) scores (PCS and MCS) to determine rhinoconjunctivitis and generic QOL. Results: Totals of 83 and 52 patients were included in the SAR and AS+SAR groups, respectively (mean age = 35.4; 56.4% females). Mini-RQLQ scores indicated slightly worse QOL in the A+SAR group at inclusion, which significantly deteriorated at the time of pollen peak, both in the SAR (p < 0.0001) and AS+SAR groups (p = 0.003). In univariate analysis, significantly higher SF-12 PCS (meaning better QOL) were observed at pollen peak in the SAR compared with the AS+SAR group (p = 0.0008), while the difference for SF-12 MCS was more limited (p = 0.05). Results were confirmed in multivariable analyses adjusting for gender, allergy medication use at pollen peak, cohort of inclusion (grass/ragweed) and comorbid conditions. Conclusions: Significant deterioration in rhinoconjunctivitis-specific QOL was observed through the pollination period in patients with SAR and AS+SAR. At pollen peak, AS+SAR patients experienced significantly worse physical functioning than patients with SAR alone.

Fas and Fas ligand expression in cystic fibrosis airway epithelium

BACKGROUND Cystic fibrosis (CF) is a genetic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene and defective expression of CFTR protein in epithelial cells. The main cause of mortality in CF is linked to chronic inflammatory and infectious airway processes. Recent studies have suggested perturbations in the apoptotic process in CF cell lines and enterocytes. A study was undertaken to investigate the expression of Fas and Fas ligand (FasL) in CF bronchial epithelium and CF tracheal cell lines. METHODS Immunohistochemical staining for Fas (alkaline phosphatase anti-alkaline phosphatase) and FasL (immunoperoxidase) was performed in eight CF bronchial epithelial samples and four controls and immunohistochemical DNA fragmentation (TUNEL) was carried out in four CF patients and four controls. Immunofluorescence staining and flow cytometric analysis of Fas and FasL expression was performed in two human tracheal epithelial cell lines (HTEC) with normal and CF genotype. The dosage of serum soluble FasL was examined in 21 patients with CF and 14 healthy volunteers. RESULTS FasL expression was markedly increased in patients with CF in both the ciliated and submucosal glandular bronchial epithelium compared with controls; Fas was similarly expressed in bronchial samples from controls and CF patients in both the ciliated epithelium and submucosal glands. High levels of DNA fragmentation were observed in CF but with some epithelial cell alterations. Serum concentrations of soluble FasL were frequently undetectable in patients with CF. In vitro, HTEC expressed Fas and FasL in both genotypes. A higher mean fluorescence intensity for FasL expression was noted in CF genotype HTEC with median (range) for six experiments of 74 (25–101) for CF cells and 42 (21–70) for non-CF cells. CONCLUSION Fas/FasL interaction is probably implicated in the human CF airway apoptotic pathway. The mechanisms of induction of FasL expression and its role in inducing tissue damage or remodelling or in controlling local inflammatory cell apoptosis remain to be determined.

Flow cytometry analysis of T lymphocytes in sarcoidosis

To examine the immunologic alterations in patients with sarcoidosis we characterized the cell cycle phase of T lymphocytes that were collected from peripheral blood and from bronchoalveolar lavage fluid. T-cell DNA and RNA contents were measured at the single cell level using flow cytometry after staining with the metachromatic fluorochrome acridine orange. T-enriched lymphocyte suspensions were obtained from peripheral blood and from bronchoalveolar lavage in 17 patients with histologically-proved sarcoidosis (10 patients, stage I and 7 patients, stage II) and in 4 patients with acute extrinsic hypersensitivity pneumonitis (AEHP). The percentages of cells in the S + (GS + M) phase in the peripheral blood of the patients with sarcoidosis did not differ from those of healthy control subjects. With bronchoalveolar lavage, however, elevated numbers of T cells in the S + (G2 + M) phase were found in the patients with AEHP and in those with stage II sarcoidosis when compared to patients with stage I sarcoidosis. The cellular RNA content of T lymphocytes from the peripheral blood showed a typical bimodal distribution without difference between patients and control subjects. Conversely, T lymphocytes obtained by bronchoalveolar lavage from patients with AEHP and sarcoidosis had a homogeneous low RNA content which differed from that of T lymphocytes from the blood from that of in vitro phytohemagglutinin-stimulated lymphocytes. These findings provide a new approach to the study of the mechanisms of local T-cell activation in sarcoidosis.

Sarcoidosis complicating anti-cytotoxic T-lymphocyte-associated antigen-4 monoclonal antibody biotherapy

To the Editor: In August 2009, a 57-yr-old male smoker (30 pack-yrs) without medical history was diagnosed as having a melanoma of the left shoulder. A complete surgical excision was performed. The lesion was ulcerated. Breslow’s thickness was 1.034 mm and Clark’s level was III. As the axillary sentinel lymph node was positive for tumour cells, an additional lymphadenectomy was performed. The disease was finally staged at IIIb (pT2b N2a M0), in agreement with the American Joint Committee on Cancer Classification. In December 2009, the patient was included in a placebo-controlled trial to evaluate ipilimumab (a monoclonal antibody anti-cytotoxic T-lymphocyte-associated antigen (CTLA)-4 antibody), after complete resection of a high-risk stage III melanoma. Ipilimumab was administered by intravenously at 10 mg·kg−1, every 3 weeks for four doses (induction) followed by 10 mg·kg−1 every 12 weeks (maintenance). After two infusions of the maintenance phase, in July 2010, subcutaneous nodules appeared on his left arm and elbow. A concomitant computed tomography (CT) scan showed multiple micronodular, reticulonodular lesions of the lung and bilateral hilar lymph nodes. Positron emission tomography–CT showed an intense fluorodeoxyglucose binding of lung nodules and mediastinal lymph nodes (fig. 1). The skin lesions were biopsied and a pathological study revealed the presence of noncaseating granulomas. Bronchoalveolar lavage showed a mild lymphocytic alveolitis (11%, with a predominance of …

Angiotensin converting enzyme in bronchoalveolar lavage fluid in pulmonary sarcoidosis.

Alveolar angiotensin converting enzyme (ACE) and serum ACE were measured simultaneously in 16 patients with histologically confirmed sarcoidosis and in 16 control subjects. Although alveolar ACE was abnormally elevated in all 15 cases of active sarcoidosis, serum ACE was not. No correlations were found between radiographic staging of pulmonary sarcoidosis and the levels of these enzymes. There was a clear correlation, however, between the levels of alveolar ACE and counts made on bronchoalveolar lavage fluid. This correlation was closer than that existing between serum ACE and bronchoalveolar lavaged lymphocytes. It is suggested that alveolar ACE is an additional biological marker of pulmonary sarcoidosis which is possibly more sensitive than serum ACE.

Quality of asthma care: results from a community pharmacy based survey

Background:  Optimal control is a major objective of disease management of asthma. The aim of the present study was to provide descriptive data on disease management in asthma patients, including medical resource utilization.

Influence of sociodemographic factors on quality of life during pollen season in seasonal allergic rhinitis patients

BACKGROUND Quality of life (QOL) is an important outcome in asthma and seasonal allergic rhinitis (SAR), and its determinants are imperfectly understood. More specifically, the influence of sociodemographic factors on QOL in patients with SAR has been so far little investigated. OBJECTIVE To examine the changes of QOL during the pollen season in patients with isolated SAR or SAR associated with asthma. METHODS A prospective cohort study was conducted in southern France. Outpatients aged 18 to 60 years and regularly treated by respiratory physicians for SAR (with or without associated asthma) were identified. Patients were recruited before the grass or ragweed pollination period. At peak pollination, patients completed the French versions of the Mini Rhino-conjunctivitis Quality of Life Questionnaire (mini-RQLQ) and the 12-item Short-Form Health Survey (SF-12) physical component summary (PCS) and mental component summary (MCS). RESULTS A total of 135 patients was included, 83 with isolated SAR and 52 with associated asthma (mean age, 35.4 years; SD, 10.6 years; 56% female). At pollen peak, QOL scores were lower in women for all instruments, with significant effects on SF-12 MCS and PCS scores in multivariate analyses. Likewise, a university-level education was an independent predictor of higher SF-12 PCS and MCS scores. Patients who lived in rural areas had significantly poorer QOL at pollen peak, as measured by the mini-RQLQ (P = .002) and SF-12 PCS (P = .008). No influence of age, presence of an animal at home, or smoking status could be identified on any QOL scores. CONCLUSIONS Being a woman, living in the countryside, and having a lower education level were all independent predictors of poorer QOL of SAR patients. These factors must be taken into account when interpreting QOL of patients with SAR. Further studies are needed to confirm these results.