PubMed · 2520563
The source did not provide an abstract. Follow the original record for more information.
Explore connections, maps & timelines
R P Sen, T E Walsh. 1989. Bronchoscopy. Enough or too much?. https://doi.org/10.1378/chest.96.4.710
Cite the original work for its findings. Save a collection to share your selection of sources.
KEEP EXPLORING
We have compared an immunocytochemical and a flow cytofluorimetric method to detect intracellular IFN-gamma, IL-4 and IL-5 in T-cell clones, peripheral blood mononuclear cells (PBMC) and bronchoalveolar lavage fluid (BALF) cells. Intracellular bound cytokine-specific antibodies were visualized either with amino-ethyl carbazole (for immunocytochemistry), or with fluorescent antibodies (for flow cytofluorimetry). The staining was inhibited with recombinant cytokines and corresponded qualitatively and quantitatively to cytokine levels in the supernatants of T-helper-0 (Th0), Th1 and Th2 clones. In analysing in vitro stimulated cells, sufficient signal in the fluorimetric assay was only obtained after the addition of monensin to the cultures. We then observed a good correlation between immunocytochemical (with no monensin added) and the flow cytofluorimetric staining for all three cytokines (PBMC, IFN-gamma and IL-4, rho = 0.9, no IL-5 detectable; clones, IL-5, rho = 0.81, all three p < 0.05). However, compared to flow cytometry, a greater percentage of positively stained cells was frequently observed using immunocytochemistry. In BALF cells, the immunocytochemical method was able to detect significant percentages of positive cells without in vitro stimulation of the cells, in contrast to the flow cytofluorimetric method. In BALF cells from sarcoidosis patients, T-cells were mainly IFN-gamma-positive (immunocytochemically assessed), both with (mean +/- SEM, 39.7 +/- 9.8%), and without (3.5 +/- 1.3%) in vitro stimulation. In BALF cells from allergic subjects, the immunocytochemical method showed lymphocytes positive for IFN-gamma (40.3 +/- 8.3%), IL-4 (19.1 +/- 0.49) and IL-5 (6.1 +/- 3.1). We conclude that both methods can be used to assess the production of IFN-gamma, IL-4 or IL-5 at the single-cell level in T-cell clones, PBMC and cells from the BALF. The high sensitivity and the low number of cells required for the immunocytochemical method indicate that this method can provide detailed information on cytokine production of airway-derived cells in diseases with airway inflammation such as sarcoidosis and asthma.
A sensitive and selective method for the determination of cefuroxime in bronchoalveolar lavage (BAL) fluid using high-performance liquid chromatography (HPLC) with UV detection at 280 nm after solid-phase extraction with C18 cartridges was developed. A Waters symmetry C18 column was used and the mobile phase was acetonitrile-0.05 M ammonium phosphate buffer (pH 3.2) (15:85, v/v). The method enabled the determination of cefuroxime at concentrations below 100 ng/ml, with a linear calibration curve at concentrations of 5-100 ng/ml for 400 microliters of BAL. The intra- and inter-assay coefficient of variations for 10, 40 and 80 ng/ml were between 5.3 and 8.9%. Analytical recoveries were between 92.7 and 106.2%. The detection limit was 1 ng/ml at a signal-to-noise ratio of 3:1 using 400 microliters of BAL. The method was successfully used for the analysis of BAL fluid from patients after oral administration of 500 mg cefuroxime axetil twice daily.
Cytomegalovirus (CMV) pneumonitis is a common opportunistic infection in lung transplant recipients. Its diagnosis usually rests on the identification of viral inclusions in lung parenchyma obtained by transbronchial biopsy, or by examination of the cytologic material obtained by bronchioloalveolar lavage (BAL). To determine whether the use of immunocytochemistry (ICC) increases the sensitivity of cytology in the diagnosis of CMV pneumonitis, we retrospectively selected 17 cases in which transbronchial biopsy and BAL were performed simultaneously, and had positive histology with negative cytology. Five negative controls were selected. The 22 slides were decolorized and restained with ICC for CMV. Of the 17 slides, nine (53%) showed cells with positive nuclear staining. All controls were negative. These results were then correlated with the number of infected cells present in the biopsy tissue, and the location of the cells (interstitial vs. intraalveolar). A good correlation was found between positive cytology and intraalveolar location of infected cells, and no correlation was seen between number of infected cells in the biopsy and the positive cytology. In summary, although histologic evaluation of lung parenchyma obtained by transbronchial biopsy is more sensitive for diagnosis of CMV pneumonitis, the sensitivity of the cytologic evaluation of BAL material can be increased by the use of ICC. The likelihood of positive ICC seems to be related to the presence of infected cells in the alveolar space rather than to the number of infected cells.