α1-antitrypsin and its C-terminal fragment attenuate effects of degranulated neutrophil-conditioned medium on lung cancer HCC cells, in vitro

BACKGROUND: Tumor microenvironment, which is largely affected by inflammatory cells, is a crucial participant in the neoplastic process through promotion of cell proliferation, survival and migration. We measured the effects of polymorphonuclear neutrophil (PMN) conditioned medium alone, and supplemented with serine proteinase inhibitor α-1 antitrypsin (AAT) or its C-terminal fragment (C-36 peptide), on cultured lung cancer cells. METHODS: Lung cancer HCC cells were grown in a regular medium or in a PMN-conditioned medium in the presence or absence of AAT (0.5 mg/ml) or its C-36 peptide (0.06 mg/ml) for 24 h. Cell proliferation, invasiveness and release of IL-8 and VEGF were analyzed by [(3)H]-thymidine incorporation, Matrigel invasion and ELISA methods, respectively. RESULTS: Cells exposed to PMN-conditioned medium show decreased proliferation and IL-8 release by 3.9-fold, p < 0.001 and 1.3-fold, p < 0.05, respectively, and increased invasiveness by 2-fold (p < 0.001) compared to non-treated controls. In the presence of AAT, PMN-conditioned medium loses its effects on cell proliferation, invasiveness and IL-8 release, whereas VEGF is up-regulated by 3.7-fold (p < 0.001) compared to controls. Similarly, C-36 peptide abolishes the effects of PMN-conditioned medium on cell invasiveness, but does not alter its effects on cell proliferation, IL-8 and VEGF release. Direct HCC cell exposure to AAT enhances VEGF, but inhibits IL-8 release by 1.7-fold (p < 0.001) and 1.4-fold (p < 0.01) respectively, and reduces proliferation 2.5-fold (p < 0.01). In contrast, C-36 peptide alone did not affect these parameters, but inhibited cell invasiveness by 51.4% (p < 0.001), when compared with non-treated controls. CONCLUSIONS: Our data provide evidence that neutrophil derived factors decrease lung cancer HCC cell proliferation and IL-8 release, but increase cell invasiveness. These effects were found to be modulated by exogenously present serine proteinase inhibitor, AAT, and its C-terminal fragment, which points to a complexity of the relationships between tumor cell biological activities and local microenvironment

Human mast cells decrease SLPI levels in type II - like alveolar cell model, in vitro.

Background Mast cells are known to accumulate at sites of inflammation and upon activation to release their granule content, e.g. histamine, cytokines and proteases. The secretory leukocyte protease inhibitor (SLPI) is produced in the respiratory mucous and plays a role in regulating the activity of the proteases. Result We have used the HMC-1 cell line as a model for human mast cells to investigate their effect on SLPI expression and its levels in cell co-culture experiments, in vitro. In comparison with controls, we found a significant reduction in SLPI levels (by 2.35-fold, p < 0.01) in a SLPI-producing, type II-like alveolar cell line, (A549) when co-cultured with HMC-1 cells, but not in an HMC-1-conditioned medium, for 96 hours. By contrast, increased SLPI mRNA expression (by 1.58-fold, p < 0.05) was found under the same experimental conditions. Immunohistochemical analysis revealed mast cell transmigration in co-culture with SLPI-producing A549 cells for 72 and 96 hours. Conclusion These results indicate that SLPI-producing cells may assist mast cell migration and that the regulation of SLPI release and/or consumption by mast cells requires interaction between these cell types. Therefore, a "local relationship" between mast cells and airway epithelial cells might be an important step in the inflammatory response

Plasma levels of alpha1-antichymotrypsin and secretory leukocyte proteinase inhibitor in healthy and chronic obstructive pulmonary disease (COPD) subjects with and without severe α1-antitrypsin deficiency

BACKGROUND: Individuals with severe Z α1-antitrypsin (AAT) deficiency have a considerably increased risk of developing chronic obstructive lung disease (COPD). It has been hypothesized that compensatory increases in levels of other protease inhibitors mitigate the effects of this AAT deficiency. We analysed plasma levels of AAT, α1-antichymotrypsin (ACT) and secretory leukocyte protease inhibitor (SLPI) in healthy (asymptomatic) and COPD subjects with and without AAT deficiency. METHODS: Studied groups included: 71 asymptomatic AAT-deficient subjects (ZZ, n = 48 and SZ, n = 23, age 31 ± 0.5) identified during Swedish neonatal screening for AAT deficiency between 1972 and 1974; age-matched controls (MM, n = 57, age 30.7 ± 0.6); older asymptomatic ZZ (n = 10); healthy MM (n = 20, age 53 ± 9.6); and COPD patients (ZZ, n = 10, age 47.4 ± 11 and MM, n = 10, age 59.4 ± 6.7). Plasma levels of SLPI, AAT and ACT were analysed using ELISA and immunoelectrophoresis. RESULTS: No significant difference was found in plasma ACT and SLPI levels between the healthy MM and the ZZ or SZ subjects in the studied groups. Independent of the genetic variant, subjects with COPD (n = 19) had elevated plasma levels of SLPI and ACT relative to controls (n = 153) (49.5 ± 7.2 vs 40.7 ± 9.1 ng/ml, p < 0.001 and 0.52 ± 0.19 vs 0.40 ± 0.1 mg/ml, p < 0.05, respectively). CONCLUSION: Our findings show that plasma levels of ACT and SLPI are not elevated in subjects with genetic AAT deficiency compared MM controls and do not appear to compensate for the deficiency of plasma AAT

Experimental and Clinical Studies of SLPI, with Special Reference to IgE-Mediated Allergic Reactions

In this study we demonstrated the production of SLPI (Secretory Leukocyte Protease Inhibitor) in serous glands in the nasal mucosa. We have shown that the pattern of the expression of mRNA corresponds to the encoded protein. The encoded protein was detected by immunohistochemical methods and mRNA was discovered by in situ hybridisation. Nasal mucosa from 11 test subjects without atopic disposition was used for this in vitro study. We found that SLPI inhibits IgE mediated histamine release in a dose dependent way. SLPI had no influence on spontaneous histamine release. Double-immunolabelling revealed that SLPI coexists with tryptase and chymase in tonsilar mast cells. The proportion SLPI/tryptase was 60% and the proportion SLPI/chymase was 37%. SLPI was found in 31% of these cells. In situ hybridisation detected SLPI mRNA in all mast cells. In situ hybridisation was performed as a double immunostaining, using a mouse anti-human mast cell antibody as mast cell identification. Mast cells in nasal mucosa also showed immunoreactivity for SLPI. We investigated the role of SLPI in patients with allergic rhinitis. From this point of view, we also examined leokocyte elastase, a1- PI and albumin. We used the method of unilateral antigen challenge. There was a higher level of SLPI in lavage fluid from healthy subjects than from atopic patients. SLPI increased in response to allergen challenged in atopics but not in healthy subjects. SLPI also increased in the non challenged left nostril, indicating that neural reflexes are involved in the SLPI-release. There was a higher concentration of elastase, a1-PI and albumin before antigen challenge in atopics patients out of pollen season than in healthy subjects. An increase in elastase, a1-PI and albumin was seen in the right challenged nostril, but not in the left non-challenged side exclusively in the atopic subjects. An irrelevant antigen did not increase the secretion of SLPI, elastase, a1- PI and albumin in atopic or healthy subjects

Päivähoidon ja hyvinvointineuvolan moniammatillinen yhteistyö päivähoidon henkilöstön kuvaamana

Opinnäytetyömme kuvailee päivähoidon ja hyvinvointineuvolan yhteistyötä päivähoidon näkökulmasta. Tuomme esiin päivähoidon työntekijöiden omia kokemuksia ja ajatuksia yhteistyöstä. Hyvinvointineuvolamallin tarkoitus on tuoda moniammatillinen apu lähelle perhettä entistä helpommin. Päivähoidon ja hyvinvointineuvolan esteetön yhteistyö edistää tavoitteiden saavuttamista ja tukee oikea-aikaista, matalan kynnyksen avunsaantia. Tutkimuksen tausta-ajatuksena on halu edistää lasten ja lapsiperheiden hyvinvointia ja ennaltaeh-käistä ongelmien syntyä. Toimeksiantajana on Oulun kaupunki. Tutkimuksemme tavoite on tuottaa kokemuksellista tietoa siitä, millainen päivähoidon ja hyvinvointineuvolan moniammatillinen yhteistyö on lastenhoitajien ja lastentarhanopettajien kuvaamana. Tutkimusaineisto koostuu neljän työntekijän, kahden lastenhoitajan ja kahden lastentarhanopettajan, teemahaastatteluista. Tutkimus kohdistuu kahteen Oulun kaupungin Koskelan alueen päiväkotiin. Kvalitatiivisen tutkimuksen aineisto analysoitiin teoriaohjaavalla sisällönanalyysilla. Teoreettisessa viitekehyksessä määritellään perheen hyvinvointi, moniammatillinen yhteistyö sekä päivähoidon ja hyvinvointineuvolan tarjoama tuki. Tutkimustuloksista ilmenee päivähoidon ja hyvinvointineuvolan yhteistyötä edistäviä ja estäviä tekijöitä. Vakiintuneita käytäntöjä ovat yhteistyöryhmä Luotsi, laajennettu nelivuotistarkastus ja lapsen perinteiset neuvolakäynnit. Ulkomaalaistaustaisten perheiden tarpeet ja lapsen kouluun siirtymisen vaihe lisäävät yhteistyötä. Työntekijät toteavat, että yhteistyö on nykyisellään vähäistä ja sitä tulisi lisätä. Haasteena nähdään salassapitosäännökset ja vaitiolovelvollisuus. Myös vähäinen tieto hyvinvointineuvolan toiminnasta koetaan esteeksi. Kuvatessaan nykyistä yhteistyötä työntekijät esittävät myös ehdotuksia yhteistyön kehittämiseksi.Our thesis describes the cooperation between a welfare clinic and day care from the perspective of day care personnel. We discuss day care staff’s experiences and views on the cooperation. The purpose of welfare clinics is to give multiprofessional aid close to families easier than before. Barrier-free cooperation between a welfare clinic and day care contributes to reaching this aim and supports families getting low-threshold help on time. Through this thesis, we hope to advance welfare in families with children and prevent problems from arising. The study was commissioned by the City of Oulu. The aim of the study was to gain experiential knowledge on how multiprofessional cooperation between a welfare clinic and day care is described by childminders and kindergarten teachers. The study focused on two kindergartens in the Koskela area in the City of Oulu. The data consisted of theme-based interviews of four employees, two of whom were childminders and two kindergarten teachers. The qualitative data was analysed by using theory-guided content analysis. The theoretical framework of the thesis presents and defines the concepts of family welfare, multiprofessional cooperation, and the support provided by welfare clinics and day care. Study results indicated factors that hindered or fostered cooperation. A cooperation team Luotsi, 4-year-olds’ extended assessments and traditional child welfare clinic visits were established cooperation practices. Close cooperation was required when working with families with a foreign background and children who are starting at school. Based on the study results, there is little cooperation currently but this was hoped to increase. Secrecy orders and professional secrecy were conceived to set challenges for cooperation. In addition, it was found that day care staff had too little information on welfare clinics’ operations. However, when describing the present state of cooperation, interviewees had ideas how to develop communication between the welfare clinic and the day care

Localisation of secretory leucocyte proteinase inhibitor mRNA in nasal mucosa

Human secretory leucocyte proteinase inhibitor (SLPI) is a low-molecular weight, acid-stable inhibitor of polymorphnuclear granulocyte elastase and cathepsin G. Previous reports have demonstrated the existence of SLPI in the respiratory tract, salivary glands and cervical mucosa. Positive staining for SLPI using immunohistochemical techniques has been reported in serous glands in nasal mucosa. We now confirm this observation and show, using in situ hybridization, that the pattern of expression of mRNA corresponds to the distribution of the encoded protein, SLPI. This, together with the high concentration of SLPI in nasal secretions, confirms the hypothesis of a local production of SLPI in the mucous membranes

Identification of SLPI (Secretory leukocyte protease inhibitor) in human mast cells using immunohistochemistry and in situ hybridisation

Recently interest has been focused on secretory leucocyte protease inhibitor (SLPI) and its role in immediate hypersensitive reactions, possibly by inhibiting mast cell chymase. The purpose of this investigation was to show whether or not SLPI is produced in mast cells. Double-immunolabelling revealed that SLPI coexists with mast cell tryptase (60%) and chymase (37%). On the other hand, in situ hybridisation studies demonstrated the expression of SLPI mRNA in all mast cells. The differences in results can be attributed to the fact that in situ hybridisation is a more sensitive method than immunohistochemistry. Hence, we conclude that SLPI is produced in human tonsillar mast cells

Human mast cells decrease SLPI levels in type II – like alveolar cell model, <it>in vitro</it>

Abstract Background Mast cells are known to accumulate at sites of inflammation and upon activation to release their granule content, e.g. histamine, cytokines and proteases. The secretory leukocyte protease inhibitor (SLPI) is produced in the respiratory mucous and plays a role in regulating the activity of the proteases. Result We have used the HMC-1 cell line as a model for human mast cells to investigate their effect on SLPI expression and its levels in cell co-culture experiments, in vitro. In comparison with controls, we found a significant reduction in SLPI levels (by 2.35-fold, p Conclusion These results indicate that SLPI-producing cells may assist mast cell migration and that the regulation of SLPI release and/or consumption by mast cells requires interaction between these cell types. Therefore, a "local relationship" between mast cells and airway epithelial cells might be an important step in the inflammatory response.</p