Search

One of the key requirements for the diagnosis of pulmonary tuberculosis is the identification of M. tuberculosis in tissue. In this paper, we present the advantages of specific fluorescent antibody labelling, combined with laser scanning confocal microscopy (LSCM), for the detection of M. tuberculosis in histological specimens of lung tissues. We demonstrate that the application of LSCM allows: (i) The automatic acquisition of images of the whole slice and, hence, the determination of regions for subsequent analysis, (ii) the acquisition of images of thick (20&ndash, 40 &mu, m) slices at high resolution, (iii) single bacteria identification, and (iv) 3D reconstruction, in order to obtain additional information about the distribution, size, and morphology of solitary M. tuberculosis, as well as their aggregates and colonies, in various regions of tuberculosis inflammation. LSCM allows for the discrimination of the non-specific fluorescence of bacteria-like particles and their aggregates presented in histological lung samples, from the specific fluorescence of labelled M. tuberculosis, using spectrum emission analysis. The applied method was effective in the identification of M. tuberculosis in lung histological samples with weak Ziehl&ndash, Neelsen staining. Altogether, combining immunofluorescent labelling with the application of LSCM visualization significantly increases the effectiveness of M. tuberculosis detection.

Goal of the study: to investigate structural and functional specific features of macrophages and cytokine profiles of bronchoalveolar lavage in new and relapsing respiratory sarcoidosis (RS). Materials and methods. The study included comparative light optical and electronic microscopic examinations of macrophages, macrophage formula and cytokine profile of bronchoalveolar lavage in 120 patients suffering from different forms of RS. It was found that in case of new RS mostly macrophages of M1 phenotype were detected in the lavage. They had high level of IL-8, IL-2, IL-1β production and ultrastructural signs of hypersecretion which was especially typical of platycytes. High level of IL-4, IL-5 content is typical of relapsing sarcoidosis: additionally to macrophages of M1 phenotype, there are macrophages of M2 phenotype with ultrastructural signs of phagocytic function which can be used for diagnostics of this form of RS.

Cells of the host containing special proteins-transporters, so-called proteins of multiple drug resistance (MDR proteins) can contribute to the drug resistance formation. Goal of the study: to define specific expression of gene and distribution of main MDR proteins (MDR1/Pgp, MRP1, lRP, BCRP) in the pulmonary cells in case of active tuberculosis. Materials and methods. Expression of MDR protein genes was evaluated by RT-PCR for mRNA isolated from the surgical specimen of fibrous cavernous tuberculosis patients. Localization of MDR proteins was performed by immunohistochemical staining and confocal laser microscopy. Main results. Intensity of MDR protein genes expression varies in different zones of tuberculous lesions: MDR1 and BCRP are characterized by the highest level and MRP1 gene is characterized by the minimum level. The level of lRP gene expression depends on the inflammation zone and it is maximum in perifocal zone where protein is detected in the cells of alveolar epithelium and macrophages. High expression of MDR protein genes in various parts of tuberculous lesions witnesses about the potential involvement of these proteins in the development of drug resistance to anti-tuberculosis drugs.