[MORPHO-FUNCTIONAL CHARACTERISTICS OF BLOOD CELL MITOCHONDRIA IN BRONCHIAL ASTHMA].

We undertook a structural and functional study of blood cell mitochondria in 25 patients with controlled mild bronchial asthma (BA) including evaluation of blood saturation with oxygen, carboxyhemoglobin level in blood and carbon monoxide content in the exhaled air. Membrane potential of leukocyte mitochondria was determined based on the results of flow cytofluorimetry and fatty acid (FA) composition in platelet mitochondrial membranes measured by GLC. It was shown that the absence of clinical symptoms of BA during remission was associated with a reduction of membrane potential and a change of FA composition resulting in the depletion of the basal pool of saturated (12:0, 14:0, 18:0) and polyunsaturated (20:4n-6, 20:5n-3, 22:5n-3, 22:4n-6) FA. These changes in the structural and functional state of blood cell mitochondria in patients with BA are signs of disordered energy-producing activity, membrane permeability and transmembrane transport suggesting the development of mitochondrial dysfunction and cellular hypoxia. A deeper insight into the role of the structural and functional state of blood cell mitochondria in the formation of respiratory disorders will facilitate early detection of the risk and complications of bronchial obstruction.

Measurement of mitochondrial membrane potential in leukocyte suspension by fluorescent spectroscopy.

A method for evaluating mitochondrial membrane potential in isolated leukocyte suspension with the use of sensitive fl uorescent cation-active dye 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine (JC-1) and spectrophotometry is described. JC-1 monomer rapidly penetrates through mitochondrial membrane of living cell and form aggregations characterized by red fl uorescence (λ = 585 nm). In case of mitochondrial membrane depolarization (early sign of apoptosis), JC-1 is not accumulated in the mitochondria and is present in the cytoplasm as a monomer characterized by green spectral fl uorescence (λ = 510 nm). The method can be used for evaluation of the function of living cells and mechanisms regulating energy metabolism by evaluating the mitochondrial membrane potential.

[Development of pulmonary chlamydia infection in inbred mice lines differentiated by genetically determinated sensitivity to tuberculosis infection].

Mice of I/St strain develop severe lung inflammation and die shortly following infection with virulent mycobacteria. The susceptibility does not depend on the Nramp1 gene, as I/St mice carry its resistant allele, but is controlled by little interacting QTL mapped to chromosomes 3, 9, 17. To find out whether the tuberculosis-susceptible I/St mice are susceptible to other intracellular bacteria taxonomically distant pathogen of Chlamydia pneumoniae was studied. Comparison of I/St and TB-resistant A/Sn mice (both Nramp1r) demonstrated that the former were more susceptible to chlamydia, displaying a significantly shortened survival time following challenge (I/St, 9.2 +/- 1.2 days; A/Sn, 22.0 +/- 0 days (p < 0.001)). To estimate the degree of chlamydial multiplication in the lungs, we suggested a quantitative real-time polymerase chain reaction (PCR)-based method which allows enumeration of the parasite's genome equivalents in infected tissue from 1 to 16 days after challenge. The interstrain difference of chlamydia burden in lungs was observed only after 24 hours after infection. Multiplication of chlamydia in the lungs was controlled efficiently after day 4 of infection. The numbers of genome equivalents dropped slightly by day 8 both in I/St and A/Sn mice. Lung pathology develops more rapidly in I/St compared to A/Sn mice following infection with chlamydia despite their similar ability to control bacterial multiplication. Lung tissue of susceptible I/St mice was markedly infiltrated with macrophages (p < 0.01), which differed significantly from the lungs of resistant A/Sn mice. In agreement with higher macrophage content in the lungs, significantly more macrophage-derived proinflammatory cytokines TNF-? and IL-6 were detected in lung tissue homogenates obtained from I/St mice (p < 0.05). Because the prominent difference in survival time did not correlate with permanent difference in bacterial multiplication, we suggested that both infections trigger fatal pathological processes whose dynamics depends strongly upon the host genetics.

[Immunomorphological characteristics of experimental granulomatous inflammation caused by Mycobacterium avium].

The results of the topical arrangement of T and B lymphocytes in the lung tissue granulomatous masses of the mice susceptible (C57BL/6Ycit) and resistant (I/StSnEgYCit) to aerosol inoculation with M. avium 724R in a dose of 2 x 10(3) CFU/mice are presented by means of labeled monoclonal antibodies. At week 8 of the inoculation, CD4+ lymphocytes were located both among the granuloma cells in the diffuse fashion and as small clusters and in the interalveolar and interlobular septa in the B6 mice and these were mainly in the granuloma in the I/St mice. At week 16, CD8+ lymphocytes were found in the peripheral granuloma portions in the B6 mice and these were diffusely throughout the area of granulomatous masses in the I/St mice. In the B6 mice, CD19+ lymphocytes formed cell aggregates from week 8 of the inoculation; and in the I/st mice, these were diffusely located among the granuloma cells and abundantly in the interalveolar and interlohular septa.