Katharina Schwarz

Characterization of Exhaled Particles from the Healthy Human Lung—A Systematic Analysis in Relation to Pulmonary Function Variables

BACKGROUND Noninvasive monitoring of airway inflammation is important for diagnosis and treatment intervention of lung disease. Mediators of interest are often nonvolatile molecules that are exhaled as aerosols and captured by breath condensation. Because analysis of exhaled breath condensate has been troublesome in the past, partly due to poor standardization and unknown dilution, we investigated in detail the influence of respiratory variables on exhaled particle number and size distribution during tidal breathing in healthy volunteers. METHODS Particle number was detected by a condensation nuclei counter, and size distribution was determined by a laser spectrometer online with high time resolution while subjects underwent a defined protocol of normal and deep tidal breathing. Intra- and intersubject variability of particle emission was analyzed and physical properties of exhaled aerosols were correlated to pulmonary function variables obtained by body-plethysmography. RESULTS The particle size distribution was in the submicron range and stable during tidal breathing. Increasing tidal volumes dominantly influenced particle number emission while flow rates had only little effect. Reproducibility within subjects was high, but there was a large variation of particle emission between subjects. The ratio of functional residual capacity to total lung capacity was found to correlate with exhaled particle numbers. This indicates that particle generation is caused by reopening of terminal airways and is dependent on functional residual capacity. CONCLUSION We conclude that online determination of exhaled aerosols from the human lungs is a prerequisite to standardize the assessment of nonvolatile mediators by normalization to the aerosol emission rate.

Functional Testing of an Inhalable Nanoparticle Based Influenza Vaccine Using a Human Precision Cut Lung Slice Technique

Annual outbreaks of influenza infections, caused by new influenza virus subtypes and high incidences of zoonosis, make seasonal influenza one of the most unpredictable and serious health threats worldwide. Currently available vaccines, though the main prevention strategy, can neither efficiently be adapted to new circulating virus subtypes nor provide high amounts to meet the global demand fast enough. New influenza vaccines quickly adapted to current virus strains are needed. In the present study we investigated the local toxicity and capacity of a new inhalable influenza vaccine to induce an antigen-specific recall response at the site of virus entry in human precision-cut lung slices (PCLS). This new vaccine combines recombinant H1N1 influenza hemagglutinin (HAC1), produced in tobacco plants, and a silica nanoparticle (NP)-based drug delivery system. We found no local cellular toxicity of the vaccine within applicable concentrations. However higher concentrations of NP (≥103 µg/ml) dose-dependently decreased viability of human PCLS. Furthermore NP, not the protein, provoked a dose-dependent induction of TNF-α and IL-1β, indicating adjuvant properties of silica. In contrast, we found an antigen-specific induction of the T cell proliferation and differentiation cytokine, IL-2, compared to baseline level (152±49 pg/mg vs. 22±5 pg/mg), which could not be seen for the NP alone. Additionally, treatment with 10 µg/ml HAC1 caused a 6-times higher secretion of IFN-γ compared to baseline (602±307 pg/mg vs. 97±51 pg/mg). This antigen-induced IFN-γ secretion was further boosted by the adjuvant effect of silica NP for the formulated vaccine to a 12-fold increase (97±51 pg/mg vs. 1226±535 pg/mg). Thus we were able to show that the plant-produced vaccine induced an adequate innate immune response and re-activated an established antigen-specific T cell response within a non-toxic range in human PCLS at the site of virus entry.

Thoracic and respirable aerosol fractions of spray products containing non-volatile compounds

ABSTRACT A versatile and simple mass balance method for the measurement of the release fraction of thoracic and respirable particles of non-volatile compounds of spray products is presented. The release fractions are defined as the ratio between the mass of suspended non-volatile particulate matter in the thoracic and respirable particle size range and the total mass of non-volatile material released with the spray action. For its determination, a spray bolus of short duration and of defined mass is sprayed into a well stirred control chamber. The respirable and thoracic aerosol mass associated with the spray bolus is determined by measuring the time averaged mass concentration inside the control volume and the half time of the exponential concentration decrease to be expected in well stirred systems to correct for mass losses during sampling.  The method is used for a wide range of spray products and technologies for which the release fractions vary by orders of magnitude. A set of data is presented elucidating the relationship between spray technology and fine particle release. Furthermore, a simple rule of thumb was derived from the data that allows for estimation of the release fractions based on a characteristic diameter of the spray droplets. The usefulness of the mass balance method for substance classification as well as for generating input data for exposure assessment and indoor air quality modeling is discussed.

Fast detection of air contaminants using immunobiological methods

The fast and direct identification of possibly pathogenic microorganisms in air is gaining increasing interest due to their threat for public health, e.g. in clinical environments or in clean rooms of food or pharmaceutical industries. We present a new detection method allowing the direct recognition of relevant germs or bacteria via fluorescence-labeled antibodies within less than one hour. In detail, an air-sampling unit passes particles in the relevant size range to a substrate which contains antibodies with fluorescence labels for the detection of a specific microorganism. After the removal of the excess antibodies the optical detection unit comprising reflected-light and epifluorescence microscopy can identify the microorganisms by fast image processing on a single-particle level. First measurements with the system to identify various test particles as well as interfering influences have been performed, in particular with respect to autofluorescence of dust particles. Specific antibodies for the detection of Aspergillus fumigatus spores have been established. The biological test system consists of protein A-coated polymer particles which are detected by a fluorescence-labeled IgG. Furthermore the influence of interfering particles such as dust or debris is discussed.

Airway and systemic inflammatory responses to ultrafine carbon black particles and ozone in older healthy subjects

ABSTRACT Increased adverse health effects in older subjects due to exposure to ambient air pollutants may be related to the inflammatory response induced by these contaminants. The aim of this study was to assess airway and systemic inflammatory responses in older healthy subjects to a controlled experimental exposure with spark-generated elemental carbon black ultrafine particles (cbUFPs) and ozone (O3). Twenty healthy subjects, age 52–75 years, were exposed on three occasions separated by at least 8 weeks. The exposures to filtered air (FA), to cbUFP (50 μg/m3), or to cbUFP in combination with 250 ppb ozone (cbUFP + O3) for 3 h with intermittent exercise were performed double blind, and in random order. Sputum and blood samples were collected 3.5 h after each exposure. Exposure to cbUFP + O3 significantly increased plasma club cell protein 16 (CC16), the number of sputum cells, the number and percent of sputum neutrophils, and sputum interleukin 6 and matrix metalloproteinase 9. Exposure to cbUFP alone exerted no marked effect, except for an elevation in sputum neutrophils in a subgroup of 13 subjects that displayed less than 65% sputum neutrophils after FA exposure. None of the inflammatory markers was correlated with age, and serum cardiovascular risk markers were not markedly affected by cbUFP or cbUFP + O3. Exposure to cbUFP+O3 induced a significant airway and systemic inflammatory response in older healthy volunteer subjects. The effects induced by cbUFP alone suggest that the inflammation was predominantly mediated by O3, although one cannot rule out that the interaction of cbUFP and O3 played a role.