Skip to main content

Main menu

  • Home
  • Current issue
  • ERJ Early View
  • Past issues
  • ERS Guidelines
  • Authors/reviewers
    • Instructions for authors
    • Submit a manuscript
    • Open access
    • Peer reviewer login
    • WoS Reviewer Recognition Service
  • Alerts
  • Subscriptions
  • ERS Publications
    • European Respiratory Journal
    • ERJ Open Research
    • European Respiratory Review
    • Breathe
    • ERS Books
    • ERS publications home

User menu

  • Log in
  • Subscribe
  • Contact Us
  • My Cart

Search

  • Advanced search
  • ERS Publications
    • European Respiratory Journal
    • ERJ Open Research
    • European Respiratory Review
    • Breathe
    • ERS Books
    • ERS publications home

Login

European Respiratory Society

Advanced Search

  • Home
  • Current issue
  • ERJ Early View
  • Past issues
  • ERS Guidelines
  • Authors/reviewers
    • Instructions for authors
    • Submit a manuscript
    • Open access
    • Peer reviewer login
    • WoS Reviewer Recognition Service
  • Alerts
  • Subscriptions

Biomonitoring for assessment of organic dust-induced lung inflammation

L. J. Mueller-Anneling, M. E. O'Neill, P. S. Thorne
European Respiratory Journal 2006 27: 1096-1102; DOI: 10.1183/09031936.06.00092204
L. J. Mueller-Anneling
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
M. E. O'Neill
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
P. S. Thorne
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Abstract

Inhalation exposure to particulate matter containing endotoxin (or lipopolysaccharide (LPS)) occurs in a variety of occupations. Nasal lavage and induced sputum have been used to evaluate lung inflammation resulting from such exposures. Whole blood assay (WBA) measures cytokine production of leukocytes after ex vivo stimulation with LPS. The present study examined the effectiveness of WBA for evaluating inflammatory responses and susceptibility.

C3HeB/FEJ mice were tolerised by LPS injection or sham tolerised with saline. Animals then inhaled either swine barn dust extract containing endotoxin or saline. Bronchoalveolar lavage (BAL) fluid was assayed for leukocyte counts and pro-inflammatory cytokines (interleukin-6, tumour necrosis factor-α). Whole blood was stimulated with 10 or 100 ng·mL-1 of LPS, incubated for 5 or 18 h and assayed for cytokines.

Barn dust-exposed groups revealed significantly higher total cells, neutrophils and cytokines in BAL compared with saline-exposed groups. Animals tolerised to LPS and exposed to barn dust demonstrated lower cellular and cytokine BAL responses. Similarly, WBA yielded significantly elevated cytokines with barn dust exposure and reduced responses with tolerisation.

This study demonstrates the efficacy of whole blood assay as a biomarker of inhalation exposure to inflammatory agents and its use for assessing susceptibility to organic dust-induced lung inflammation.

  • Biomonitoring
  • endotoxin
  • exposure assessment
  • organic dust
  • whole blood assay

Organic dust is a complex bio-aerosol consisting of bacterial and fungal products and their metabolites, animal, insect, and plant components. Inhalation of organic dust may cause severe acute pulmonary and systemic inflammation and lead to occupational asthma and other obstructive pulmonary diseases 1, 2. Studies have shown that a nonallergic, neutrophil-mediated response accounts for these pulmonary health outcomes and that pro-inflammatory cytokines, such as tumour necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6, play a major role in the inflammatory process 2–4. A significant component of organic dust responsible for lung inflammation is endotoxin, also referred to as lipopolysaccharide (LPS). Endotoxin exposure is associated with adverse respiratory effects in the indoor environment 5 and is a well-recognised occupational hazard in: 1) swine, poultry and dairy barns; 2) grain handling facilities; 3) vegetable and cotton processing; 4) sawmills, metal machining, fibreglass production operations; and 5) composting and waste handling 6. Endotoxin forms the exterior layer of the outer cell membrane of nearly all Gram-negative bacteria and is known to have potent pro-inflammatory effects attributed to the lipid-A region of the molecule 7–9. Since this region is imbedded in the bacterial cell membrane, endotoxin is most potent when released by intact cells or upon disruption of the membrane 10.

Studies have demonstrated that a decline in forced expiratory volume in one second following endotoxin or organic dust exposure is associated with increased production of inflammatory mediators by alveolar macrophages and peripheral blood monocytes 11. It may therefore be possible to evaluate immune responses to organic dust-induced airway inflammation through analysis of the ex vivo release of cytokines by blood monocytes or alveolar macrophages. However, isolating and culturing these cells is time consuming and not conducive to studies of multiple subjects. In addition, isolated cells may not respond in the same manner as when maintained in their natural matrix. Although not a perfect substitute, an ex vivo whole blood assay (WBA) attempts to preserve various blood cell interactions necessary for cytokine response and cell viability and may be a suitable alternative 12, 13. Unlike nasal lavage or induced sputum production, the volume of blood used for the assay is invariable, which allows for the determination of accurate total cell counts. Several implementations of whole blood analysis have been used to examine immune responses to various agents and conditions, including pyrogens 14–16, sepsis 17, 18, infectious disease 17, 19, 20, allergens 21, 22 and other environmental agents 23–25. Using methods similar to this study, Wouters et al. 26 assessed within- and between-subject variation of the WBA in normal, unexposed volunteers and found that there was relatively low within-subject variance compared with between-subject variance, particularly for IL-1β and IL-6. Intra-individual variances for these two cytokines at LPS stimuli of 12.5 and 100 ng·mL-1 were 0.15 and 0.18 for IL-1β and 0.14 and 0.12 for IL-6, respectively. As with other studies, the whole blood responded in a dose–response relationship to the inflammatory agents LPS and curdlan. Of the four cytokines analysed, the greatest consistency of response was observed with IL-1β and IL-6 26.

Previous research in the present authors’ laboratory demonstrated that mice tolerised to endotoxin prior to organic dust exposure had a reduced pulmonary inflammatory response upon inhalation exposure to grain dust as compared to their naïve counterparts 27. This phenomenon has been previously reported in the literature 28 and has been referred to as endotoxin or LPS “tolerance” or “adaptation” 2, 27, 29, 30. The present study evaluates WBA response as a biomarker of exposure to organic dust and uses the endotoxin-tolerised mouse as a model for workers tolerised to endotoxin through daily occupational exposure. These experiments sought to test whether mice exposed to organic dust from swine confinement facilities have a more vigorous whole blood cytokine response to endotoxin than sham-exposed animals and if the cytokine response of mouse whole blood stimulated ex vivo increases with higher concentrations of endotoxin stimulant 31. A second objective was to determine whether LPS tolerance led to reduced responsiveness as assessed using WBA.

MATERIALS AND METHODS

Animals

Male, 6-week-old C3HeB/FeJ mice (Jackson Labs, Bar Harbor, ME, USA) were housed in an American Association for Accreditation of Laboratory Animal Care-accredited rodent vivarium. All protocols were reviewed and approved by the Institutional Animal Care and Use Committee of the University of Iowa (Iowa City, IA, USA). Mice were quarantined for 12 days before the experiments were initiated. Four mice were reserved as sentinels and necropsied at the beginning and end of the experiment to verify animal health. Thirty-two mice were divided into two exposure groups: a sham exposure group, which received nebulised saline; and an organic dust exposure group, which was exposed to a nebulised extract of dust collected from a concentrated animal feeding operation (CAFO) housing swine (CAFO dust). The strain of mouse chosen for the current study has been previously characterised as “endotoxin sensitive” in an organic dust exposure model 27.

Induced lipopolysaccharide tolerance

Endotoxin tolerance was induced in half of the mice from each exposure group, as previously described 27, 32. Briefly, mice received daily intraperitoneal (i.p.) injections of increasing doses of Escherichia coli O111:B4 endotoxin (Sigma, St Louis, MO, USA) as follows. Day 1: 100 μg·kg-1; day 2: 500 μg·kg-1; day 3: 1,000 μg·kg-1; day 4: 5,000 μg·kg-1. Endotoxin concentration of the stock solution was confirmed by kinetic chromogenic Limulus amebocyte lysate (LAL) assay (BioWhittaker Inc., Walkersville, MD, USA). The remaining 16 control mice were administered pyrogen-free saline in a similar fashion.

Swine CAFO dust extract

Dust samples were obtained from an Eastern Iowa swine CAFO by vacuuming vertical surfaces with a sampling vacuum. An extract of the dust was prepared at a concentration of 100 mg·mL-1 in sterile, physiological saline. The dust was eluted by first vortexing for 2 min and then shaking on a laboratory rotator for 1 h at ambient temperature. Insoluble particles were removed by centrifuging and decanting twice at 3,350×g for 15 min each. The final supernatant was adjusted to pH 7.2.

Exposure

Mice were exposed in a whole-body exposure chamber 33 for 4 h to either nebulised saline (n = 16) or nebulised CAFO dust extract (n = 16). In each exposure group, half of the mice were from the induced tolerance (LPS i.p.) group and the remaining were controls (saline i.p.). Aerosols were generated with a six-jet Collison nebuliser (BGI Inc., Waltham, MA, USA) with a 138-kPa air supply 7. Airflow through the chamber was regulated by an exhaust flow of 22.5 L·min-1. Passive airflow into the exposure chamber was filtered through an organic vapour cartridge and P100 particulate filter (North Safety Products, Cranston, RI, USA). Endotoxin concentration within the exposure chamber was measured by hourly collection of 15-min air samples collected on 47-mm glass fibre filters.

Bronchoalveolar lavage fluid and whole blood collection

At 1 h post-exposure (5 h after the initiation of the inhalation exposure), animals were anaesthetised, euthanised and exsanguinated by cardiac puncture to obtain heparinised whole blood. Blood from all animals of an exposure group was pooled and held at 4°C. Pooling of blood from individual mice was necessary to obtain sufficient blood for the WBA. Immediately following blood collection, bronchoalveolar lavage fluid (BALF) was obtained for total and differential cell counts and cytokine analyses. The lungs were lavaged with sterile, pyrogen-free saline at a pressure of 25 cmH2O in 1-mL increments for a total volume of 4 mL. BALF was stored at 4°C and processed as soon as possible after collection. The BALF was centrifuged for 5 min at 200×g and the resulting supernatant was decanted, divided into equal volume aliquots and frozen at -80°C for later cytokine analysis. The remaining cell pellet was re-suspended in RPMI media and prepared for total and differential cell counts. Total counts were performed using an improved Neubauer hemocytometer (Reichert, Buffalo, NY, USA), and the Diff Quick Stain Set (Harleco, Gibbstown, NY, USA) was used for staining for differential enumeration by microscopy.

Whole blood assay

Whole blood samples were pooled by endotoxin tolerance and exposure groups, and diluted with an equal volume of saline. The diluted whole blood was pipetted into tissue culture tubes in duplicate aliquots of 900 μL. To each of these tubes, 100 μL of LPS was added to stimulate cytokine release as follows: high LPS stimulant (100 ng·mL-1), low LPS stimulant (10 ng·mL-1), and saline control. LPS was purchased from BioWhittaker Inc. as a lyophilised extract of E. coli serotype O55:B5. Tubes with blood and stimulant were incubated at 37°C at 95% humidity and 5% carbon dioxide in duplicate. Following either a 5- or 18-h incubation, 500 μL saline was added to each tube before centrifugation at 1,000×g for 15 min. The resulting plasma supernatants were decanted and stored at -80°C until cytokine analysis.

ELISA and Limulus amebocyte lysate assays

Murine IL-6 and TNF-α cytokine concentrations in bronchoalveolar lavage (BAL) supernatant and WBA plasma samples were determined using commercially available ELISA kits (BioSource International, Camarillo, CA, USA). Endotoxin concentrations of air filter samples were determined using kinetic chromogenic LAL assay, performed as previously described 7.

Statistical methods

Primary dependent variables included BAL total and differential cell counts, BALF cytokine concentrations, and WBA cytokine production with and without LPS stimulation. For the WBA, net induced cytokine production was calculated by subtracting the nonstimulated control from the corresponding LPS stimulated result. Unpaired t-tests were used to analyse BALF data. Comparisons were considered significant at p≤0.01.

RESULTS

Exposure

The concentration of the endotoxin delivered in the CAFO dust extract exposure averaged 28.4±4.1 μg·m-3. This concentration corresponds to a body burden of 490 ng·kg-1 (4,900 EU·kg-1), which is ∼10 times the lung burden a CAFO worker would amass over an 8-h working day in a CAFO operated without any exposure controls 34.

Results of bronchoalveolar lavage fluid analysis

Significantly higher total and neutrophil cell counts as well as cytokine concentrations were observed for both CAFO dust-exposed groups as compared with the sham-exposed groups (p<0.001; fig. 1⇓). Mice exposed to CAFO dust had a far higher concentration of total BAL cells than the sham-exposed group mean (p<0.001; fig. 1a⇓). This response was marked by an influx of neutrophils, resulting in an increase from 1% neutrophils in controls to 97% in CAFO dust-exposed mice. Figure 1b⇓ illustrates that neutrophils increased in concentration from 230 cells·mL-1 in controls to 1,600,000 cells·mL-1 in the CAFO dust-exposed mice.

Fig. 1—
  • Download figure
  • Open in new tab
  • Download powerpoint
Fig. 1—

a) Total cells, b) neutrophils, c) interleukin (IL)-6 and tumour necrosis factor (TNF)-α in bronchoalveolar lavage fluid (BALF) from sham-tolerised (No tls) and tolerised (lipopolysaccharide (LPS) tls) mice exposed to saline (bars shaded green) and concentrated animal feeding operation dust (bars shaded yellow); sentinels are represented by orange shading. Error bars indicate mean±sem. **: p<0.01; ***: p<0.001.

Animals tolerised to LPS and exposed to CAFO dust had significantly lower BALF total cells (1.13×106 versus 1.61×106 cells·mL-1; p<0.001) and neutrophils (1.08×106 versus 1.59×106 cells·mL-1; p<0.001) compared with animals that received the same exposure but without tolerisation (fig. 1a⇑ and b). Furthermore, as shown in figure 1c⇑, tolerised animals in the CAFO dust-exposed group did not have as vigorous a pulmonary inflammatory response as the nontolerised animals, as measured by IL-6 release (3,400 versus 1,100 pg·mL-1; p<0.001). There was no difference in TNF-α between these two groups (fig. 1d⇑). However, the TNF-α response was extremely high (>17,000 pg·mL-1) for both the tolerised and nontolerised groups exposed to CAFO dust. There were no significant differences in pulmonary inflammatory responses between the control (sham-exposed) tolerised and nontolerised groups.

Whole blood assay

Results of the WBA on pooled blood from the animals described previously are shown in figure 2⇓. The whole blood was incubated for either 5 or 18 h with no in vitro stimulation or stimulation with low or high amounts of LPS. In most cases, there was an increased release of cytokines in vitro with increasing endotoxin stimulation. Cytokine release was higher in non-tolerised CAFO dust-exposed mice than sham-exposed mice at the 5- and 18-h time points for IL-6 but not TNF-α. Tolerising animals to LPS prior to CAFO dust-extract exposure dramatically reduced their IL-6 response compared with the exposed, nontolerised animals for both the 5- and 18-h time points. As with BALF, this difference was not generally reflected in the sham-exposed groups. The pattern of responsiveness between exposure (sham versus CAFO dust) and tolerance (saline versus LPS) groups for TNF-α suggested a stronger response to LPS stimulation in the animals that were not exposed to CAFO dust. This may be due to the fact that TNF-α response among mice exposed to CAFO dust peaked before the 5-h time point.

Fig. 2—
  • Download figure
  • Open in new tab
  • Download powerpoint
Fig. 2—

Whole blood analysis (WBA) cytokine results for a) interleukin (IL)-6 and b) tumour necrosis factor (TNF)-α following 5-h incubation of the whole blood after lipopolysaccharide (LPS) stimulation and c) IL-6 and d) TNF-α following 18-h incubation. WBA from the saline (green bars) and concentrated animal feeding operation (CAFO; yellow) groups were either unstimulated (lightest bars), stimulated with low LPS (mid-shaded bars) or stimulated with high LPS (darkest bars). No tls: nontolerised; LPS tls: tolerised.

DISCUSSION

Mice exposed to inhaled swine CAFO dust developed a profound lung inflammatory response as demonstrated by a 40-fold increase in BALF cells (30-fold for the tolerised group), a 6,500-fold increase in neutrophils (8,500 for tolerised group) and a ∼250-fold increase in two cytokines that serve as markers of inflammation: TNF-α and IL-6. Sham-exposed mice who breathed nebulised, pyrogen-free saline did not demonstrate any changes in these outcome variables as compared with sentinel animals.

The present study demonstrated that mice tolerised to endotoxin prior to inhalation exposure to swine CAFO dust have significantly fewer total cells and neutrophils in BALF. These mice also responded less vigorously in WBA than nontolerised mice receiving the same exposure. Differences in susceptibility between exposure groups were evident, as demonstrated by the greater WBA IL-6 cytokine responses in both CAFO dust-exposed groups as compared with the sham-exposed groups. The same effect was found to a lesser degree for TNF-α after 5-h WBA incubation but not 18-h incubation. It is possible this was due to the timing of the assay, since TNF-α production peaks much earlier than IL-6 13. Furthermore, in human WBA studies, TNF-α production has been found to be much more variable both within and between individuals 12, 26. These data indicate that WBA functions as a biomarker assay of exposure effect and of reduced susceptibility due to induced tolerance, and suggest that IL-6 might be a more reliable marker of this effect.

Endotoxin exposures were ∼10-fold higher than would be experienced by CAFO workers 6, 34. Despite this difference in exposure used to elicit a response, WBA might also be applicable to human inhalation exposure studies. It is known that individuals vary in their response to inhalation of organic dusts and LPS in both occupational and experimental settings 11, 35. The nature of this difference may relate to factors in the pathway of signal transduction, from endotoxin to toll-like receptor (TLR)4 via LPS-binding protein, CD14 and MD2 36. Another significant factor may be genetic determinants of immune responses to LPS. TLR4 polymorphisms may account for some of this variability, but other genes also seem to play an important role 37–39. Arbour et al. 40 found that two common TLR4 mutations (Asp299Gly and Thr399Ile) were associated with a decreased response to inhaled LPS in humans. Insertion of the wild-type allele into primary airway epithelial cell or alveolar macrophage cell cultures from individuals with the aforementioned TLR4 mutations could reverse this effect in vitro. Some of this variability may also depend on the type of endotoxin (cell-bound versus purified) present in the exposure 41 and pre-existing conditions such as asthma 42. A whole blood in vitro stimulation assay attempts to account for this variability by maintaining an “intact” cellular milieu; it may therefore be a good predictor of an individual's immune response to various environmental and microbial agents 43.

Workers exposed to high endotoxin in CAFOs, in grain-dust or composting facilities, or in vegetable-washing operations may become tolerised to endotoxin. One way in which this is apparent is in “Monday morning fever” in cotton and textile workers, where those exposed to endotoxin early in the week (i.e. after a weekend off from work) have more significant respiratory symptoms than they do later in the week 44. Data from this study suggest that cytokine production in the WBA for cotton would diminish over the course of the working week.

Several previous studies have sought to validate WBA 15, 16, 26, 45. Wouters et al. 26 determined that between-individual variability in WBA with LPS and β-glucan stimulation was more significant than within-individual variability. WBA was a reliable, reproducible measure of an individual's responsiveness. However, that study used unexposed normal subjects so no inference could be made regarding the utility of WBA for biomonitoring exposures to inflammatory agents. Other studies have characterised LPS-induced cytokine responses of cultured cells, including monocytes and lymphocytes, but did not investigate WBA 13, 39, 46. The role of occupationally induced LPS tolerance in WBA has not been adequately explored.

In conclusion, the present study in a murine inhalation exposure model suggests that whole blood analysis may serve as a biomarker of exposure and an alternative to various methods that are either nonspecific or more invasive and not applicable to large, population-based occupational exposure studies. Further research is needed in human exposure models to confirm these findings. One such study is currently underway in the present authors’ laboratory.

Acknowledgments

The authors gratefully acknowledge the helpful suggestions of G. Doekes of Utrecht University (Utrecht, the Netherlands).

  • Received August 4, 2004.
  • Accepted February 13, 2006.
  • © ERS Journals Ltd

References

  1. ↵
    American Thoracic Society. Respiratory health hazards in agriculture. Am J Respir Crit Care Med 1998;158:S1–S76.
    OpenUrlCrossRefPubMedWeb of Science
  2. ↵
    Rylander R. Endotoxin in the environment - exposure and effects. J Endotoxin Res 2002;8:241–252.
    OpenUrlCrossRefWeb of Science
  3. Clapp WD, Becker S, Quay J, et al. Grain dust-induced airflow obstruction and inflammation of the lower respiratory tract. Am J Respir Crit Care Med 1994;150:611–617.
    OpenUrlCrossRefPubMedWeb of Science
  4. ↵
    Jagielo PJ, Thorne PS, Watt JL, Frees KL, Quinn TJ, Schwartz DA. Grain dust and endotoxin inhalation challenges produce similar inflammatory responses in normal subjects. Chest 1996;110:263–270.
    OpenUrlCrossRefPubMedWeb of Science
  5. ↵
    Thorne PS, Kulhankova K, Yin M, Cohn R, Arbes SJ, Zeldin DC. Endotoxin exposure is a risk factor for asthma: the national survey of endotoxin in United States housing. Am J Respir Crit Care Med 2005;172:1371–1377.
    OpenUrlCrossRefPubMedWeb of Science
  6. ↵
    Douwes J, Thorne P, Pearce N, Heederik D. Bioaerosol health effects and exposure assessment: progress and prospects. Ann Occup Hyg 2003;47:187–200.
    OpenUrlAbstract/FREE Full Text
  7. ↵
    Thorne PS. Inhalation toxicology models of endotoxin- and bioaerosol-induced inflammation. Toxicology 2000;152:13–23.
    OpenUrlCrossRefPubMedWeb of Science
  8. Wiese A, Brandenburg K, Ulmer AJ, Seydel U, Muller-Loennies S. The dual role of lipopolysaccharide as effector and target molecule. Biol Chem 1999;380:767–784.
    OpenUrlCrossRefPubMedWeb of Science
  9. ↵
    Thorn J. The inflammatory response in humans after inhalation of bacterial endotoxin: a review. Inflamm Res 2001;50:254–261.
    OpenUrlCrossRefPubMedWeb of Science
  10. ↵
    Khan SA, Everest P, Servos S, et al. A lethal role for lipid A in Salmonella infections. Mol Microbiol 1998;29:571–579.
    OpenUrlCrossRefPubMedWeb of Science
  11. ↵
    Kline JN, Cowden JD, Hunninghake GW, et al. Variable airway responsiveness to inhaled lipopolysaccharide. Am J Respir Crit Care Med 1999;160:297–303.
    OpenUrlCrossRefPubMedWeb of Science
  12. ↵
    Ojeda Ojeda M, Silva CV, de J Arana Rosainz M, Fernandez-Ortega C. TNFalpha production in whole blood cultures from healthy individuals. Biochem Biophys Res Commun 2002;292:538–541.
    OpenUrlCrossRefPubMedWeb of Science
  13. ↵
    Allen JN, Herzyk DJ, Allen ED, Wewers MD. Human whole blood interleukin-1-beta production: kinetics, cell source, and comparison with TNF-alpha. J Lab Clin Med 1992;119:538–546.
    OpenUrlPubMedWeb of Science
  14. ↵
    Poole S, Mistry Y, Ball C, et al. A rapid “one-plate” in vitro test for pyrogens. J Immunol Methods 2003;274:209–220.
    OpenUrlCrossRefPubMedWeb of Science
  15. ↵
    Fennrich S, Fischer M, Hartung T, et al. Detection of endotoxins and other pyrogens using human whole blood. Dev Biol Stand 1999;101:131–139.
    OpenUrlPubMed
  16. ↵
    Hartung T, Wendel A. [Detection of pyrogens using human whole blood]. ALTEX 1995;12:70–75.
    OpenUrlPubMed
  17. ↵
    Rigato O, Salomao R. Impaired production of interferon-gamma and tumor necrosis factor-alpha but not of interleukin 10 in whole blood of patients with sepsis. Shock 2003;19:113–116.
    OpenUrlCrossRefPubMedWeb of Science
  18. ↵
    Yaqub S, Solhaug V, Vang T, et al. A human whole blood model of LPS-mediated suppression of T cell activation. Med Sci Monit 2003;9:BR120–BR126.
    OpenUrlPubMed
  19. ↵
    House D, Chinh NT, Hien TT, et al. Cytokine release by lipopolysaccharide-stimulated whole blood from patients with typhoid fever. J Infect Dis 2002;186:240–245.
    OpenUrlAbstract/FREE Full Text
  20. ↵
    Diterich I, Harter L, Hassler D, Wendel A, Hartung T. Modulation of cytokine release in ex vivo-stimulated blood from borreliosis patients. Infect Immun 2001;69:687–694.
    OpenUrlAbstract/FREE Full Text
  21. ↵
    Ellaurie M, Yost SL, Rosenstreich DL. A simplified human whole blood assay for measurement of dust mite-specific gamma interferon production in vitro. Ann Allergy 1991;66:143–147.
    OpenUrlPubMedWeb of Science
  22. ↵
    Kruger T, Sigsgaard T, Bonefeld-Jorgensen EC. Ex vivo induction of cytokines by mould components in whole blood of atopic and non-atopic volunteers. Cytokine 2004;25:73–84.
    OpenUrlCrossRefPubMedWeb of Science
  23. ↵
    Sigsgaard T, Bonefeld-Jorgensen EC, Kjaergaard SK, Mamas S, Pedersen OF. Cytokine release from the nasal mucosa and whole blood after experimental exposures to organic dusts. Eur Respir J 2000;16:140–145.
    OpenUrlAbstract/FREE Full Text
  24. Fennrich S, Zucker B, Hartung T. [A new application for the human whole blood test: development of an assay to assess the health risk of air-borne microbial contaminations]. ALTEX 2001;18:41–46.
    OpenUrlPubMedWeb of Science
  25. ↵
    Bønløkke JH, Thomassen M, Viskum S, Omland O, Bonefeld-Jorgensen E, Sigsgaard T. Respiratory symptoms and ex vivo cytokine release are associated in workers processing herring. Int Arch Occup Environ Health 2004;77:136–141.
    OpenUrlCrossRefPubMedWeb of Science
  26. ↵
    Wouters IM, Douwes J, Thorne PS, Heederik D, Doekes G. Inter- and intraindividual variation of endotoxin- and beta(1 --> 3)-glucan-induced cytokine responses in a whole blood assay. Toxicol Ind Health 2002;18:15–27.
    OpenUrlAbstract/FREE Full Text
  27. ↵
    Schwartz DA, Thorne PS, Jagielo PJ, White GE, Bleuer SA, Frees KL. Endotoxin responsiveness and grain dust-induced inflammation in the lower respiratory tract. Am J Physiol 1994;267:L609–L617.
  28. ↵
    Snella MC, Rylander R. Lung cell reactions after inhalation of bacterial lipopolysaccharides. Eur J Respir Dis 1982;63:550–557.
    OpenUrlPubMedWeb of Science
  29. ↵
    Elder AC, Finkelstein J, Johnston C, Gelein R, Oberdorster G. Induction of adaptation to inhaled lipopolysaccharide in young and old rats and mice. Inhal Toxicol 2000;12:225–243.
    OpenUrlPubMedWeb of Science
  30. ↵
    Beeson PB. With the technical assistance of Elizabeth Roberts. Tolerance to bacterial pyrogens: I. Factors influencing its development. J Exp Med 1947;86:29–38.
    OpenUrlAbstract
  31. ↵
    Mueller-Anneling L, O'Neill ME, Doekes G, Thorne PS. Organic dust-induced lung inflammation and the whole blood assay. Am J Respir Crit Care Med 2003;167:A760
    OpenUrlCrossRef
  32. ↵
    Coffee KA, Halushka PV, Ashton SH, Tempel GE, Wise WC, Cook JA. Endotoxin tolerance is associated with altered GTP-binding protein function. J Appl Physiol 1992;73:1008–1013.
    OpenUrlAbstract/FREE Full Text
  33. ↵
    O'Shaughnessy PT, Achutan C, O'Neill ME, Thorne PS. A small whole-body exposure chamber for laboratory use. Inhal Toxicol 2003;15:251–263.
    OpenUrlCrossRefPubMedWeb of Science
  34. ↵
    Duchaine C, Thorne PS, Meriaux A, Grimard Y, Whitten P, Cormier Y. Comparison of endotoxin exposure assessment by bioaerosol impinger and filter-sampling methods. Appl Environ Microbiol 2001;67:2775–2780.
    OpenUrlAbstract/FREE Full Text
  35. ↵
    Jacobs RR, Boehlecke B, van Hage-Hamsten M, Rylander R. Bronchial reactivity, atopy, and airway response to cotton dust. Am Rev Respir Dis 1993;148:19–24.
    OpenUrlPubMedWeb of Science
  36. ↵
    Gioannini TL, Teghanemt A, Zhang D, et al. Isolation of an endotoxin-MD-2 complex that produces Toll-like receptor 4-dependent cell activation at picomolar concentrations. Proc Natl Acad Sci USA 2004;101:4186–4191.
    OpenUrlAbstract/FREE Full Text
  37. ↵
    Lorenz E, Jones M, Wohlford-Lenane C, et al. Genes other than TLR4 are involved in the response to inhaled LPS. Am J Physiol Lung Cell Mol Physiol 2001;281:L1106–L1114.
    OpenUrlAbstract/FREE Full Text
  38. Yang RB, Mark MR, Gray A, et al. Toll-like receptor-2 mediates lipopolysaccharide-induced cellular signalling. Nature 1998;395:284–288.
    OpenUrlCrossRefPubMedWeb of Science
  39. ↵
    Hermann C, von Aulock S, Graf K, Hartung T. A model of human whole blood lymphokine release for in vitro and ex vivo use. J Immunol Methods 2003;275:69–79.
    OpenUrlCrossRefPubMedWeb of Science
  40. ↵
    Arbour NC, Lorenz E, Schutte BC, et al. TLR4 mutations are associated with endotoxin hyporesponsiveness in humans. Nat Genet 2000;25:187–191.
    OpenUrlCrossRefPubMedWeb of Science
  41. ↵
    Rylander R, Bake B, Fischer JJ, Helander IM. Pulmonary function and symptoms after inhalation of endotoxin. Am Rev Respir Dis 1989;140:981–986.
    OpenUrlPubMedWeb of Science
  42. ↵
    Michel O, Duchateau J, Sergysels R. Effect of inhaled endotoxin on bronchial reactivity in asthmatic and normal subjects. J Appl Physiol 1989;66:1059–1064.
    OpenUrlAbstract/FREE Full Text
  43. ↵
    Schins RP, van Hartingsveldt B, Borm PJ. Ex vivo cytokine release from whole blood. A routine method for health effect screening. Exp Toxicol Pathol 1996;48:494–496.
    OpenUrlPubMedWeb of Science
  44. ↵
    Rylander R. Health effects of cotton dust exposures. Am J Ind Med 1990;17:39–45.
    OpenUrlPubMedWeb of Science
  45. ↵
    Kirchner H, Kleinicke C, Digel W. A whole-blood technique for testing production of human interferon by leukocytes. J Immunol Methods 1982;48:213–219.
    OpenUrlCrossRefPubMedWeb of Science
  46. ↵
    Koyama S, Sato E, Nomura H, et al. The potential of various lipopolysaccharides to release monocyte chemotactic activity from lung epithelial cells and fibroblasts. Eur Respir J 1999;14:545–552.
    OpenUrlAbstract/FREE Full Text
View Abstract
PreviousNext
Back to top
View this article with LENS
Vol 27 Issue 6 Table of Contents
European Respiratory Journal: 27 (6)
  • Table of Contents
  • Index by author
Email

Thank you for your interest in spreading the word on European Respiratory Society .

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Biomonitoring for assessment of organic dust-induced lung inflammation
(Your Name) has sent you a message from European Respiratory Society
(Your Name) thought you would like to see the European Respiratory Society web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Print
Citation Tools
Biomonitoring for assessment of organic dust-induced lung inflammation
L. J. Mueller-Anneling, M. E. O'Neill, P. S. Thorne
European Respiratory Journal Jun 2006, 27 (6) 1096-1102; DOI: 10.1183/09031936.06.00092204

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero

Share
Biomonitoring for assessment of organic dust-induced lung inflammation
L. J. Mueller-Anneling, M. E. O'Neill, P. S. Thorne
European Respiratory Journal Jun 2006, 27 (6) 1096-1102; DOI: 10.1183/09031936.06.00092204
del.icio.us logo Digg logo Reddit logo Technorati logo Twitter logo CiteULike logo Connotea logo Facebook logo Google logo Mendeley logo
Full Text (PDF)

Jump To

  • Article
    • Abstract
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • Acknowledgments
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
  • Tweet Widget
  • Facebook Like
  • Google Plus One

More in this TOC Section

  • Involvement of MMP-12 and phosphodiesterase type 4 in cigarette smoke-induced inflammation in mice
Show more Original Articles: Lung inflammation

Related Articles

Navigate

  • Home
  • Current issue
  • Archive

About the ERJ

  • Journal information
  • Editorial board
  • Press
  • Permissions and reprints
  • Advertising

The European Respiratory Society

  • Society home
  • myERS
  • Privacy policy
  • Accessibility

ERS publications

  • European Respiratory Journal
  • ERJ Open Research
  • European Respiratory Review
  • Breathe
  • ERS books online
  • ERS Bookshop

Help

  • Feedback

For authors

  • Instructions for authors
  • Publication ethics and malpractice
  • Submit a manuscript

For readers

  • Alerts
  • Subjects
  • Podcasts
  • RSS

Subscriptions

  • Accessing the ERS publications

Contact us

European Respiratory Society
442 Glossop Road
Sheffield S10 2PX
United Kingdom
Tel: +44 114 2672860
Email: journals@ersnet.org

ISSN

Print ISSN:  0903-1936
Online ISSN: 1399-3003

Copyright © 2023 by the European Respiratory Society