PT  - JOURNAL ARTICLE
AU  - Mikhail Vladimirsky
AU  - Marina Lapenkova
AU  - Yulia Alyapkina
AU  - Irina Vasilyeva
TI  - Efficiency of bactericidal activity of liposomal mycobacteriophages against intracellular Mycobacteria tuberculosis in the model of macrophages RAW 264
AID  - 10.1183/13993003.congress-2019.PA4607
DP  - 2019 Sep 28
TA  - European Respiratory Journal
PG  - PA4607
VI  - 54
IP  - suppl 63
4099  - http://erj.ersjournals.com/content/54/suppl_63/PA4607.short
4100  - http://erj.ersjournals.com/content/54/suppl_63/PA4607.full
SO  - Eur Respir J2019 Sep 28; 54
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).