RT Journal Article
SR Electronic
T1 Efficiency of bactericidal activity of liposomal mycobacteriophages against intracellular Mycobacteria tuberculosis in the model of macrophages RAW 264
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP PA4607
DO 10.1183/13993003.congress-2019.PA4607
VO 54
IS suppl 63
A1 Mikhail Vladimirsky
A1 Marina Lapenkova
A1 Yulia Alyapkina
A1 Irina Vasilyeva
YR 2019
UL http://erj.ersjournals.com/content/54/suppl_63/PA4607.abstract
AB Introduction: The use of lytic mycobacteriophage D29 incorporated into liposomes may be one way to achieve more efficient TB treatment.Aim: To study the bactericidal effect of liposomal lytic mycobacteriophage (phage) against intracellular MTB in the model of cultured macrophages.Methods: RAW 264-7 (ATCC) macrophage cell culture was infected for 24 hours with H37RV MTB at a concentration of 107/ml. Then macrophages were isolated by centrifugation in a Ficoll-Paque density gradient. Phage D29 (minimum 108 PFU) was purified on an ion-exchange Q-sepharose column and then shaken with a phospholipid film consisting of phosphatidylcholine and cholesterol, followed by 20-fold extrusion of the obtained liposomal mycobacteriophage particles using 0.4 microns pores filters. The liposomal inclusion of phage was controlled by analyzing phage DNA by real-time PCR and transmission electron microscopy. The experimental wells of macrophage cells, 3 for each experiment, were analyzed: control, phage and liposomal phage. After 24-hour incubation the macrophages were removed with scrubbers from the surface of the wells and after double freezing and thawing were sown on plates with agar medium 7H10 and cultivated in a CO2 incubator at 37°C.Results: Resulting counting of MTB colonies: control - 62±2,6; mycobacteriophage -17±1; liposomal mycobacteriophage -7±0,3. P1-2 <0,01; P2-3 < 0,05.Conclusions: Method of preparation macrophage culture with intracellular MTB infection was presented and the higher bactericidal efficacy of mycobacteriophages included in 400 nm liposomes was shown.FootnotesCite this article as: European Respiratory Journal 2019; 54: Suppl. 63, PA4607.This is an ERS International Congress abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).