Studies of the Specific Activity of Aerosolized Isoniazid against Tuberculosis in a Mouse Model.

The aerosol inhalation delivery of isoniazid in mice was investigated, and the specific activity of the aerosol form of isoniazid was studied with the mouse model of tuberculosis infection, the M. tuberculosis H37Rv strain. Aerosol delivery was performed using a laminar-flow horizontal nucleation chamber. The inhalation dose was measured in real-time mode using a diffusion aerosol spectrometer. The mean particle diameter was 0.6 ± 0.03 µm, and the inhalation dose was 5-9 mg/kg. Pharmacokinetic measurements were carried out in nose-only and whole-body chambers. Isoniazid concentration in blood serum and its mass in the lungs were measured as a function of time using high-performance liquid chromatography. Studies of the specific activity of aerosolized isoniazid reveal that treatment with the aerosol lead to the complete recovery of the experimental tuberculosis infection as early as after 28 days after the start of inhalation treatment, while in the animals from the group receiving isoniazid per-orally, sole revivable tuberculosis mycobacteria were detected. Histologic examinations show that only a few macrophagal (nonspecific) granulomas without mycobacteria were detected in the spleen after per-oral and aerosol treatment, the number of granulomas on the 28th day being three times smaller in the latter case. The results show that the developed technique of isoniazid aerosol inhalation may have clinical potential.

Modeling of Mycobacterium tuberculosis dormancy in bacterial cultures.

The currently available methods are unable to directly detect dormant forms of Mycobacterium tuberculosis (Mtb) in vivo. The persistence of Mtb in the host body is detectable only in an indirect manner via the immunological response to Mtb-specific antigens. It is commonly recognized that the pathogen prevalently exists in the human body in a latent stage. Additional research efforts focusing on the Mtb dormancy are needed for development of sterilizing drugs, which are necessary to control LTBI and stop TB epidemic. To this end, the in vitro models of Mtb dormancy may be useful. This review briefly describes the phenomenon of Mtb dormancy and its role in the context of tuberculosis as a persistent bacterial infection; then the article characterizes in details the in vitro methods used for modeling the Mtb dormancy in bacterial cultures.

BCG infection in mice is promoted by naïve mesenchymal stromal cells (MSC) and suppressed by poly(A:U)-conditioned MSC.

Mesenchymal stromal cells (MSC) transplantation is an actively studied therapeutic approach used in regenerative medicine and in the field of control of immunoinflammatory response. Conditioning of MSC in culture can form their predominantly pro- or anti-inflammatory phenotypes. We demonstrated that poly(A:U)-conditioning of bone marrow-derived mouse MSC induced predominantly pro-inflammatory phenotype. The effects of administration of naïve MSC (nMSC) or conditioned MSC (cMSC) on the course of mycobacterial infection were studied. BALB/c mice infected i.p. with 5 × 106 M. bovis BCG were successively injected i.v. with 0.75 × 106 of nMSC or cMSC in 11 and 12.5 weeks after infection and sacrificed at the week 14. Histological and bacteriological examination of BCG-infected animals revealed low bacterial loads in liver, lungs and spleen; the bacterial load in spleen was higher than in other organs. Treatment with nMSC induced 3-fold increase of the number of bacteria in spleen granulomas, while cMSC decreased significantly the number of bacteria in BCG-positive granulomas. Analysis of preparations of organ homogenates by luminescent microscopy, MGIT cultures and CFU count on Lowenstein-Jensen medium revealed that nMSC promoted mycobacterial growth whereas cMSC suppressed mycobacterial growth significantly. We concluded that MSC therapy can be effective in mycobacterial infection, but only in a case of appropriate conditioning of the cells.