Study of cell death kinds and stages of leukocytes apoptosis in pulmonary tuberculosis patient during antimycobacterial therapy
Journal Title: Туберкульоз, легеневі хвороби, ВІЛ-інфекція - Year 2019, Vol 0, Issue 2
Abstract
Objective — to evaluate the condition of cytoplasmic membranes of leukocytes and to study their viability, to determine the varieties and stages of cell death of leukocytes in patients with pulmonary tuberculosis (TB) with different treatment efficacy in the intensive phase (IP) of antimycobacterial therapy (AMBT). Materials and methods. The study involved 30 patients with an established diagnosis of pulmonary TB who were treated at the Regional Tuberculosis Hospital N 1 (Kharkiv). Patients were divided into groups depending on the effectiveness of treatment in the IP AMBT. The first group included 12 patients with effective treatment — after 60 doses of IP AMBT conversion of bacteriaexcretion and positive dynamics according to the results of clinical and radiological studies were determined. The second group included 18 patients with a delayed and/or negative result of treatment — after 60—90—120 doses of IP AMBT persistant bacteriaexcretion was detected and stable or negative dynamics were determined and/or treatment failure was established. The control group consisted of 12 healthy donors. The study was performed by flow cytometry on a flow cytometer FACS Calibur. Assessment of cells death stages was performed by adding markers CD45+, Annexin V FITC detection KIT I, 7AAD. Results and discussion. The proportion of live intact leukocytes in the blood of patients with pulmonary TB was (71.05 ± 3.66) %, compared with the control group (91.8 ± 0.1) %, which is less for 20.75 %. In the blood of TB patients an increasing of damaged cells was determined at the initial stage of apoptosis ((14.7 ± 1.47) vs (3.8 ± 0.16) % in the control group) and at the late stage of apoptosis/necrosis ((12.35 ± 2.36) vs (3.08 ± 0.3) % in the control group), which indicates a decrease in the active leukocyte pool. In the blood of patients with pulmonary TB, the proportion of dead necrotic cells decreases in 1.8 times ((0.95 ± 0.07) vs (1.78 ± 0.32) % in the control group), which may indicate the inclusion of compensatory mechanisms of the immune system. Each indicator studied was significantly (p < 0.05) different between groups. Comparing the stages of cell death in groups with different treatment efficiencies in IP AMBT showed that in each of the analyzed conditions of leukocytes, no significant (p > 0.05) differences between the comparison groups were identified, with significant (p < 0.05) differences with control group. Conclusions. TB is accompanied by profound impaired immune response, since externalization of phospholipid phosphatidylserine in the phospholipid bilayer of the cytoplasmic membrane of peripheral blood leukocytes is observed. The proportion of alive and functionally well-off leukocytes is 20.75 % less in TB patients than in practically healthy donors. Since the study was conducted on the background of AMBT, the reason for such a deficiency of cells of the leukocyte pool can be not only the progression of TB, but also the use of AMBT, which requires further study. Significant differences between the states of cytoplasmic membranes of leukocytes in groups of patients with TB with different efficacy of treatment in IP AMBT is not defined. (27.1 ± 1.9) % of peripheral blood leukocytes in patients with pulmonary TB were in a state of apoptosis, of which more than half (14.7 ± 1.47) % in the initial stage of apoptosis, indicating a therapeutic need for using additional immunomodulating drugs with the purpose of quantitative and qualitative recovery of immunocompetent cells.
Authors and Affiliations
O. S. Shevchenko, O. A. Nakonechna, O. О. Hovardovska
Keywords
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- EP ID EP674026
- DOI 10.30978/TB2019-2-50
- Views 86
- Downloads 0
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