In vitro model of latent mycobacterial infection

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/02 (2006.01) C12N 5/0786 (2010.01) C12N 1/20 (2006.01) C12Q 1/18 (2006.01)

Patent

CA 2685746

A method of inducing latency in Mycobacterium permits preparation of an in vitro model system of latent mycobacterial infection. Latency is induced in a pure culture of Mycobacterium by exposing it to multiple stress conditions, including a low nutrient culture medium without glycerol, a low pH, a relatively high level of carbon dioxide and a relatively low gas phase oxygen level. An in vitro model of mycobacterial infection employs macrophages induced from THP1 cells which are then infected with Mycobacterium. The infected macrophages are grown under hypoxic conditions to induce latency in the mycobacteria. The in vitro model of infection is useful in evaluating compounds for activity against latent mycobacteria.

L'invention concerne un procédé d'induction d'une latence dans une mycobactérie qui permet la préparation d'un système de modèle in vitro d'infection mycobactérienne latente. La latence est induite dans une culture pure de mycobactérie en l'exposant à de multiples conditions de contrainte, comprenant un milieu de culture à faible teneur en nutriment sans glycérol, un faible pH, un taux de dioxyde de carbone relativement élevé et un taux d'oxygène en phase gazeuse relativement faible. Un modèle in vitro d'infection mycobactérienne emploie des macrophages induits à partir de cellules THP1 qui sont ensuite infectés avec la mycobactérie. Les macrophages infectés croissent dans des conditions hypoxiques pour induire une latence dans les mycobactéries. Le modèle in vitro d'infection est utile dans l'évaluation des composés pour une activité par rapport aux mycobactéries latentes.

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