Cathelicidin related antimicrobial peptide, laminin, Toll-like receptors and chemokines levels in experimental hypersensitivity pneumonitis in mice.

INTRODUCTION: Hypersensitivity pneumonitis (HP) is an interstitial lung disease caused by unresolved inflammation and tissue repair pathologies triggered by repeated organic dust exposure. The aim of the study was to investigate changes in levels of the cathelicidin related antimicrobial peptide (CRAMP), laminin (LAM-A1), selected Toll-like receptors (TLR) and chemokines in experimental HP in mice. MATERIALS AND METHODS: Three and 18-month-old female C57BL/6J mice underwent inhalations of the saline extract of Pantoea agglomerans cells, Gram-negative bacterium common in organic dust and known for its pathogenic impact. The inhalations were repeated daily (28 days). ELISA was used for measuring in lung tissue homogenates concentration of CRAMP, LAM-A1, TLR2, TLR4, TLR8, CXCL9 (chemokine [C-X-C motif] ligand) and CXCL10. RESULTS: Levels of TLR2, TLR4 and CXCL9 were significantly higher in both young and old mice lungs already after 7 days of inhalations, while significant increase of LAM-A1 and CXCL10 was noted after 28 days, compared to untreated samples. TLR8 level was significantly augmented only in young mice. Only CRAMP level significantly declined. Significantly higher TLR8 and CXCL9 concentration in untreated samples were noted in old animals compared to young ones. CONCLUSION: Significant alterations of the examined factors levels indicate their role in HP pathogenesis.

Cathelicidin LL-37 in bronchoalveolar lavage and epithelial lining fluids from healthy individuals and sarcoidosis patients.

Sarcoidosis is a granulomatous disease of unknown etiology most often characterized by pulmonary manifestations. Changes in an innate immune system, involving antimicrobial peptides, have been noted during the course of pulmonary sarcoidosis. This study focuses on the level of LL-37 peptide, the only human cathelicidin, additionally characterized by a wide range of pleiotropic activities, in pulmonary sarcoidosis. A cross-sectional study was conducted in groups of 32 patients with sarcoidosis and 12 healthy individuals. Bronchoalveolar lavage fluid (BALF) sampling, followed by LL-37 measurements by mass spectrometry combined with previous immunoaffinity purification, was performed. Based on urea levels, concentrations of LL-37 in epithelial lining fluid (ELF) were calculated. The levels of LL-37 peptide in BALF samples derived from patients with pulmonary sarcoidosis (median: 17.45 pg/ml, 25th-75th percentile: 8.05-28.33 pg/ml) were significantly higher compared to the healthy group (median: 6.38 pg/ml, 25th-75th percentile: 4.90-11.55 pg/ml) (U Mann-Whitney test, p=0.04). Assessment of LL-37 in ELF confirmed the differences across the groups that were observed in BALF. The level of LL-37 in patients with sarcoidosis (median: 2.25 ng/ml, 25th-75th percentile: 1.03-5.06 ng/ml) was again higher compared to healthy individuals (median: 0.62 ng/ml, 25th-75th percentile: 0.43-2.17 ng/ml) (p=0.06, Mann-Whitney U test). The results of this study demonstrate that the level of LL-37 peptide is elevated in pulmonary compartment affected by sarcoidosis. This might have a meaning in the pathomechanism of the disease, especially taking into consideration versatile activity of human cathelicidin revealed in numerous experimental studies during the last years.

Relationship between COPD and lower socioeconomic status in farmers from South-Eastern Poland (Lublin region).

INTRODUCTION: Lower socioeconomic status is considered to be an independent risk factor of chronic conditions, such as chronic obstructive pulmonary disease (COPD). COPD, one of the major public health problems worldwide, is a chronic inflammatory lung disease of a multifactorial background. COPD morbidity in rural areas has been higher than in urban settings, as apart from the major causative factor, tobacco smoking, the burden of this disease in rural environments is also connected to additional occupational factors (organic dusts). The management of chronic diseases seems to be particularly difficult in rural areas. The aim of the study was to analyze the socioeconomic status of farmers suffering from COPD in comparison to healthy farmers. METHODS: Thirty farmers with COPD and 34 healthy farmers from the Lublin region (Poland) were investigated based on the area of land they possessed (an indicator used in the health insurance system in Poland to classify farmers). The farmers from five rural communes were selected by general practitioners. Statistical analysis was performed by non-parametric Mann-Whitney U-test for the differences between area of farms. The p < 0.05 level was considered as significant. RESULTS: Area of land (median: 1.5 ha, 25th-75th percentile: 1.0-4.0) owned by farmers with COPD was significantly lower than area of farms belonging to healthy farmers (median: 7.0 ha, 25th-75th percentile: 3.0-10.0) (p < 0001, Mann-Whitney U-test). CONCLUSIONS: In rural areas individuals with COPD are characterized by significantly lower socioeconomic status than healthy persons. COPD is a major health problem, especially in rural areas, which may indicate that policy-makers should consider addressing equity in COPD management in rural areas.

Cathelicidin LL-37 in bronchoalveolar lavage and epithelial lining fluids from COPD patients and healthy individuals.

Innate immunity is currently under scope of interest concerning its role in the development of chronic obstructive pulmonary disease (COPD). Antimicrobial peptides constitute a potent part of this fast response system. Here, we focus on the role of a specific antimicrobial peptide, the only human cathelicidin, the pleiotropic LL-37 peptide, in the development of COPD under clinical conditions. A cross-sectional study was conducted in groups of 43 patients with COPD (previously classified according to GOLD) and 12 healthy individuals. Bronchoalveolar lavage fluid (BALF) sampling, followed by LL-37 measurements by mass spectrometry combined with previous immunoaffinity purification, was performed. Based on urea levels, concentrations of LL-37 in epithelial lining fluid (ELF) were calculated. Additionally, an antimicrobial assay of growth inhibition of two bacterial species, often involved in COPD development mechanisms, by purchased LL-37 was conducted. Altogether, 55 BALF samples were analyzed. LL-37 levels were significantly higher in BALF from patients in early stages of COPD (GOLD I-II) compared to BALFs from healthy individuals. The same was true for ELF. Cathelicidin’s concentration was significantly lower in both BALF and ELF from patients in advanced COPD (GOLD III-IV). The significantly elevated LL-37 levels both in BALF and ELF in patients with COPD at stage GOLD I-II together with reduced levels in advanced (COPD stage III-IV) further supports the innate immunity involvement in COPD pathology and suggests a profound change in non-specific immunity during the disease progression.

Response of herb processing workers to work-related airborne allergens.

A group of 51 herb processing workers employed in a big herb processing facility located in eastern Poland were examined by the skin and precipitin tests with, respectively, 4 and 17 extracts of microorganisms associated with organic dusts. Out of this number, 32 workers were examined by the skin test with 7 extracts of selected herbs processed in the facility. All the subjects were asked about the occurrence of work-related symptoms. 32 healthy office workers were examined with microbial extracts as a reference group. The herb processing workers showed a high proportion of early skin reactions (after 20 min) to the extract of Gram-negative bacterium Alcaligenes faecalis (41.2%), significantly higher compared to the reference group (p<0.01). At all time intervals (20 min, 8 hrs, 24 hrs), the workers responded with a high frequency to the extract of Bacillus subtilis (respectively 72.5%, 64.7%, and 15.7%), significantly greater compared to the reference group (respectively p<0.001, p<0.001, and p<0.05). No significant differences were found between the groups of herb processing workers and referents in skin response to the extracts of Streptomyces albus and Alternaria alternata and, except for the extract of Pantoea agglomerans, in the frequency of positive precipitin reactions to microbial antigens. In the skin test with herb extracts, the highest response among workers were caused by the extracts of chamomile flowers and nettle leaves which evoked 40-65% of positive skin reactions at all time intervals. 39 out of 51 interviewed herb processing workers (76.5%) reported the occurrence of work-related general, respiratory and skin symptoms. The positive skin reactions occurred more frequently among symptomatic workers which suggests that the specific immunologic response might be implicated in etiopathogenesis of work-related symptoms in examined workers. However, in most cases the differences did not attain a significance level which indicates that there is no direct relationship between a positive immunologic response and the appearance of symptoms caused by occupational exposure to herb dust, and that most probably a considerable part of these symptoms might be also due to non-specific immunologic and/or toxic mechanisms.

Effects of exposure to flax dust in Polish farmers: work-related symptoms and immunologic response to microbial antigens associated with dust.

Medical examinations were performed in a group of 51 Polish farmers heavily exposed to flax dust during harvesting and scutching (threshing) and in a group of 50 healthy urban dwellers not exposed to organic dusts (controls). The examinations included: interview concerning the occurrence of respiratory disorders and work-related symptoms, physical examination, X-ray examination of chest, lung function tests, oxymetric examinations, determination of the concentration of cytokines (IL-1alpha IL-6, TNFalpha) in blood serum and allergological tests with microbial antigens associated with organic dust, comprising: skin prick test with 4 antigens, agar-gel precipitation test with 12 antigens and test for specific inhibition of leukocyte migration with 4 antigens. As many as 32 farmers (62.7%) reported the occurrence of work-related symptoms during harvesting, transporting and scutching of flax. The most common complaint was general weakness reported by 15 farmers (29.4%), followed by headache reported by 14 (27.5%), blocking of the nose - by 11 (21.6%), dry cough, shivering, and eyes itching - each by 10 (19.8%), chest tightness and hoarseness - each by 9 (17.6%). No control subjects reported these work-related symptoms. The mean spirometric values in the examined group of farmers were within a normal range and did not show a significant post-shift decline. In contrast, a significant post-shift decline of oxymetric values was found among flax farmers. The farmers showed a frequency of the positive early skin reactions to environmental allergens in the range of 0-19.6%, a frequency of positive precipitin reactions in a range of 0-56.9%, and frequency of positive reactions of leukocyte migration inhibition in a range of 7.8-21.6%. The members of the control group responded to the majority of allergens with a significantly lower frequency of positive results compared to the farmers. Elevated concentrations of IL-1alpha and IL-6, but not TNFalpha, were found in blood sera of flax farmers. In conclusion, farmers engaged in harvesting and scutching of flax represent a group of elevated professional risk because of high incidence of work-related symptoms and high frequency of allergic reactions to bacteria and fungi associated with organic dust.

Allergic alveolitis due to herb dust exposure.

We report an episode of allergic alveolitis in a female farmer due to massive exposure to organic dust contaminated with microorganisms during threshing of herbs (thyme). The patient's medical history, the results of exposure test, inhalation challenge, and bronchoalveolar lavage suggested the diagnosis of allergic alveolitis

Effects of exposure to grain dust in Polish farmers: work-related symptoms and immunologic response to microbial antigens associated with dust.

Medical examinations were performed in a group of 76 Polish farmers heavily exposed to grain dust during harvesting and threshing, and in a group of 63 healthy urban dwellers not exposed to organic dusts (controls). The examinations included: interview concerning the occurrence of respiratory disorders and work-related symptoms, physical examination, lung function tests, and allergological tests comprising skin prick test with 4 microbial antigens associated with grain dust and agar-gel precipitation test with 12 microbial antigens. As many as 34 farmers (44.7%) reported the occurrence of work-related symptoms during harvesting and threshing. The most common was dry cough reported by 20 individuals (26.3%). Dyspnoea was reported by 15 farmers (19.7%), tiredness by 12 (15.7%), chest tightness by 8 (10.5%), plugging of nose and hoarseness by 5 each (6. 5%). No control subjects reported these work-related symptoms. The mean spirometric values in the examined group of farmers were within the normal range, but a significant post-shift decrease of these values was observed after work with grain. The farmers showed a frequency of the positive early skin reactions to environmental allergens in the range of 10.8 - 45.5%, and a frequency of positive precipitin reactions in range of 3.9 - 40.8%. The control group responded to the majority of allergens with a significantly lower frequency of positive results compared to the farmers. The obtained results showed a high response of grain farmers to inhalant microbial allergens and indicate a potential risk of occupational respiratory diseases (such as allergic alveolitis, asthma, Organic Dust Toxic Syndrome) among this population

Study on exposure of pig farm workers to bioaerosols, immunologic reactivity and health effects.

Occupational inhalation of organic dust may be a cause of numerous symptoms and diseases. Organic dust consists of various biological compounds which induce inflammatory reactions in the lungs on an immunotoxic or allergic basis. Bacteria and their metabolites, moulds and their spores, mycotoxins, glucans, and other still unidentified substances, can be potential aetiologic factors of diseases caused by organic dust. The aim of this study was to determine the concentration and composition of airborne microflora in typical Polish swine buildings and to assess the health conditions of the workers employed within them. Total concentration of microorganisms in the air of five examined swine breeding farms ranged from 613.7-1246.7 x 10(3) cfu/m3 (mean value 930.6 x 10(3) cfu/ m3). The examination of 53 employees working in the swine buildings included their medical history, physical examination, spirometry, and allergological tests. Work-related symptoms were reported by 31 (58.5%) of the subjects. No abnormal findings were present upon physical and spirometric examinations. The results suggest the common occurrence of work-related respiratory disease in swine workers, mostly corresponding to the Organic Dust Toxic Syndrome (ODTS).

[Exposure to organic dust and microorganisms as a factor affecting respiratory function of workers of purebred horse farms]. / Narazenie na pyl organiczny i drobnoustroje jako czynnik wplywajacy na stan ukladu oddechowego pracowników stadniny koni.

In two farms of purebred horses, microbiological studies of the air and medical examination of the workers were performed. The concentration of microorganisms in the air were within the range 29.2-336.0 cfu x 10(3)/m3. The respirable fraction in most cases was between 30-60% of the total count. The levels of dust and endotoxin were low except for one sampling point where the concentration of endotoxin was as high as 3.44 g/m3. As many as 16 workers out of the total of 31 examined reported occurrence of work-related symptoms. However, the results of physical examination and of lung function tests were within normal range. A high proportion of workers showed a positive skin response to Saccharopolyspora rectivirgula (51.6%) and the presence of precipitins to Acinetobacter calcoaceticus (32.3%). No significant relationship could be found between the presence of symptoms and positive allergological reactions. Total medical results indicate the probability of the occurrence of Organic Dust Toxic Syndrome in high proportion of the workers.